WorldWideScience

Sample records for tk litwin ms

  1. Reliability analysis of Litwin and Stringer's Organizational Climate ...

    African Journals Online (AJOL)

    OCQ) developed by Litwin and Stringer (1968) in a Nigerian sample are presented. Data were collected from 600 employees of shoe, textile and food manufacturing industries in Lagos, Nigeria. An alpha co-efficient of .70 was observed for the ...

  2. Ojediran, TK

    African Journals Online (AJOL)

    Ojediran, TK. Vol 9, No 1-2 (2013) - Articles Biochemical and Heamatological Indices of Broiler Chickens fed Differently Processed Legume Seed Meals Abstract PDF · Vol 9, No 1-2 (2013) - Articles Lesions in Broiler Chicks Following Experimental Contamination with Battery Waste Abstract PDF. ISSN: 0331-5428.

  3. Perl/Tk Pocket Reference

    CERN Document Server

    Lidie, Stephen

    1998-01-01

    The Perl/Tk Pocket Reference is a companion volume to Learning Perl/Tk, an O'Reilly Animal Guide. Learning Perl/Tk is a tutorial for Perl/Tk, the extension to Perl for creating graphical user interfaces. With Tk, Perl programs can be window-based rather than command-line based, with buttons, entry fields, listboxes, menus, scrollbars, balloons, tables, dialogs, and more. And Perl/Tk programs run on UNIX and Windows-based computers. This small book is a handy reference guide geared toward the advanced Perl/Tk programmer. Novice Perl/Tk programmers will find that its compact size gives th

  4. Klubová identita TK Most

    OpenAIRE

    Čechová, Karolína

    2015-01-01

    Title: Corporate Identity of TK Most Objectives: The objective of this work is to create the list of recommendations that lead to improvement of corporate identity of TK Most. Methods: Method of non-structured interviews with players of the club, SWOT analysis. dates analysis and participant observation made by myself were used for corporate identity analysis and subsequently proposed recommendations. Results: It was revealed that corporate identity of TK Most has a couple of weaknesses. It n...

  5. Improved Separation of Pa from Th and U in Marine Sediments with TK400 Resin.

    Science.gov (United States)

    Süfke, Finn; Lippold, Jörg; Happel, Steffen

    2018-01-16

    Protactinium-231 is a radionuclide of broad interest in paleoceanography and paleoclimatology. This study describes an improved method for the purification and separation of Pa from marine sediment samples using the new TK400 resin by TrisKem International. The focus lies on the improvement of the separation of the abundant 232 Th from the Pa fraction of the sample, which would reduce the peak tailing from 232 Th on masses 231 and 233 during ICP-MS measurement. Furthermore, the reusability of TK400 has been tested. For this purpose, the conventional method using Dowex AG1X8 for the separation and purification of Pa has been compared to methods using the TK400 resin. A combination of a Dowex AG1X8 prior to a TK400 column has shown most convincing results. Based on our results we suggest a new efficient procedural method of analyzing 231 Pa from marine sediment samples using TK400. Chemical Pa yields for a Dowex-TK400 combination are highest compared to the Dowex only method. Furthermore, the 232 Th/ 231 Pa ratio of the Pa-fractions has been reduced by 1 order of magnitude compared to conventional methods with Dowex AG1X8. Additionally, the reusability of TK400 resin up to nine times has been proven. The usage of TK400 is only limited in the presence of samples with a high matrix load (e.g., Fe). Therefore, matrix from sediment samples needs to be removed (here using Dowex resin) before samples are loaded onto TK400. We also report a series of concentration measurements from standard reference materials (UREM-11, NIST 2702), which have been used for 233 Pa calibration.

  6. Tcl/Tk in a Nutshell

    CERN Document Server

    Raines, Paul

    2009-01-01

    The Tcl language and Tk graphical toolkit are powerful building blocks for custom applications. This quick reference briefly describes every command and option in the core Tcl/Tk distribution, as well as the most popular extensions. Keep it on your desk as you write scripts, and you'll be able to quickly find the particular option you need.

  7. Tcl/Tk A Developer's Guide

    CERN Document Server

    Flynt, Clif

    2011-01-01

    Newly updated with over 150 pages of material on the latest Tcl extensions, Tcl/Tk: A Developer's Guide is a unique practical tutorial for professional programmers and beginners alike. Starting with a clear picture of the basics, Tcl/Tk covers the variety of tools in this "Swiss army knife" of programming languages, giving you the ability to enhance your programs, extend your application's capabilities, and become a more effective programmer. This updated edition covers all of the new features of version 8.6, including object-oriented programming and the creation of megawidgets, existing d

  8. Experiences in effective use of Tcl/Tk

    Energy Technology Data Exchange (ETDEWEB)

    Lee, R.W.

    1995-06-01

    Tcl/Tk (Toot Command Language and Tool Kit, pronounced ``tickle tee-kay``) is a scripting language supporting Motifm style X Window interfaces. It is extendible, allowing developers to embed additional functionality as commands in the language. However, the power and flexibility of the system leads to many variations or possibilities in its usage. We describe effective methods for taking advantage of Tcl/Tk to increase productivity and enhance the flexibility and adaptability of applications: writing simple Tcl/Tk scripts, extending the Tcl/Tk widget set, wrapping Tcl commands around existing classes and functions, and building Tcl/Tk and 3GL coprocesses. Examples are presented from working applications.

  9. Implementing Portfolios Using Tk20: An Educational Assessment System

    Science.gov (United States)

    Zhang, Jie; Fallon, Moira A.; Wright, Allison M.

    2016-01-01

    The purpose of this paper is to share results of collaborative effort introducing special education portfolios into an inclusive teacher education program using the Tk20 assessment system. Tk20 is an assessment system for both providing evidence of educational skills and achieving that evidence in such a way as to demonstrate growth of teacher…

  10. Identity Management ToolKit (IdM TK)

    Data.gov (United States)

    Department of Veterans Affairs — With the IdM TK, authorized users can search and view identity and exception information from the Administrative Data Repository (ADR). Specifically, users can view...

  11. Discovery of Macrocyclic Pyrimidines as MerTK-Specific Inhibitors.

    Science.gov (United States)

    McIver, Andrew L; Zhang, Weihe; Liu, Qingyang; Jiang, Xinpeng; Stashko, Michael A; Nichols, James; Miley, Michael J; Norris-Drouin, Jacqueline; Machius, Mischa; DeRyckere, Deborah; Wood, Edgar; Graham, Douglas K; Earp, H Shelton; Kireev, Dmitri; Frye, Stephen V; Wang, Xiaodong

    2017-02-03

    Macrocycles have attracted significant attention in drug discovery recently. In fact, a few de novo designed macrocyclic kinase inhibitors are currently in clinical trials with good potency and selectivity for their intended target. In this study, we successfully engaged a structure-based drug design approach to discover macrocyclic pyrimidines as potent Mer tyrosine kinase (MerTK)-specific inhibitors. An enzyme-linked immunosorbent assay (ELISA) in 384-well format was employed to evaluate the inhibitory activity of macrocycles in a cell-based assay assessing tyrosine phosphorylation of MerTK. Through structure-activity relationship (SAR) studies, analogue 11 [UNC2541; (S)-7-amino-N-(4-fluorobenzyl)-8-oxo-2,9,16-triaza-1(2,4)-pyrimidinacyclohexadecaphane-1-carboxamide] was identified as a potent and MerTK-specific inhibitor that exhibits sub-micromolar inhibitory activity in the cell-based ELISA. In addition, an X-ray structure of MerTK protein in complex with 11 was resolved to show that these macrocycles bind in the MerTK ATP pocket. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Operability test procedure for the TK-900 effluent monitoring station

    International Nuclear Information System (INIS)

    Weissenfels, R.D.

    1994-01-01

    This procedure will verify that the 221-B liquid effluent monitoring system, installed near the east end of the 6-in. chemical sewer header, functions as intended by design. TK-900B was installed near stairwell 3 in the 221-B electrical gallery by Project W-007H. The system is part of BAT/AKART for the BCE liquid effluent system

  13. PROSES REPRESENTASI SIMBOL MATEMATIKA PADA PROSES BERMAIN ANAK TK

    Directory of Open Access Journals (Sweden)

    Ari Kusuma Sulyandari

    2017-01-01

    Full Text Available This study aims to determine the representation of children in kindergartens of the objects and symbols of numbers 1 to 10 when playing activities. So the mathematical activity in appropriate with the child's development. This study uses qualitative descriptive by Moleong. The collection of data through observation, interviews, photo and recording. To check the validity of researchers used data triangulation of data sources, theory and methodology. Results of the study is that children need visual in process representation. They need to understand the concept of visualization when many objects, counting objects, understand the numbers 1 to 10. Children are not able to think abstractly. Representation of children has not been perfect. Method of learning in kindergarten helps learning to count and recognize numbers. Especially if done with playing. Tujuan penelitian ini untuk mengetahui proses representasi anak TK pada simbol matematika bilangan asli 1 sampai 10 pada proses bermain sehingga pembelajaran matematika sesuai dengan perkembangan anak. Metode penelitian yang digunakan adalah deskriptif kualitatif. Teknik pengumpulan data menggunakan teknik observasi, wawancara, dan dokumentasi berupa foto serta perekaman. Teknik analisis data yang digunakan pada saat penelitian adalah analisis deskriptif untuk mengecek keabsahan peneliti menggunakan data triangulasi sumber data, teori, dan metodologi. Hasil penelitian menunjukkan bahwa anak usia TK membutuhkan visual dalam aktivitas representasi. Anak-anak membutuhkan bantuan visual saat memahami konsep banyak benda, menghitung benda, memahami bilangan 1 sampai 10. Hal ini dikarenakan anak usia TK masih belum dapat berpikir secara abstrak sehingga representasi anak usia TK masih belum sempurna. Penggunaan metode pembelajaran tematik terpadu di TK mempermudah pembelajaran berhitung dan mengenal angka terlebih jika dilakukan dengan bermain.

  14. ( ) 100 dk sin cTk ikX

    Indian Academy of Sciences (India)

    GENERAL I ARTICLE a function of IX I. Using eikX = cos kX + i sin A~X, we see that the contribution from the sin kX term vanishes because the integrand is an odd function of k. Thus u(1)(X, T). CB(T). -- sin (cTk) cos kX. 100 dk o 7rk. 100 dk. C B ( T ). -. ( sin (CT + X) k + sin (CT - X) k) o 27rk i B ( T) (c (CT + X) + c (CT - X) ).

  15. FISH comets show that the salvage enzyme TK1 contributes to gene-specific DNA repair.

    Science.gov (United States)

    McAllister, Katherine A; Yasseen, Akeel A; McKerr, George; Downes, C S; McKelvey-Martin, Valerie J

    2014-01-01

    Thymidine kinase 1 (TK1) is a salvage enzyme that phosphorylates thymidine, imported from surrounding fluids, to create dTMP, which is further phosphorylated to the DNA precursor dTTP. TK1 deficiency has for a long time been known to cause increased cellular sensitivity to DNA damage. We have examined preferential strand break repair of DNA domains in TK1(+) and TK1(-) clones of the Raji cell line, by the Comet-FISH technique, in bulk DNA and in the actively transcribed tumor suppressor (TP53) and human telomerase reverse transcriptase (hTERT) gene regions, over 1 h after 5Gy γ-irradiation. Results showed that repair of the TP53 and hTERT gene regions was more efficient in TK1(+) compared to TK1(-) cells, a trend also reflected to a lesser degree in genomic DNA repair between the cell-lines. The targeted gene-specific repair in TK(+) cells occurred rapidly, mainly over the first 15 min repair-period. Therefore, TK1 is needed for preferential repair of actively transcribed regions, through a previously unsuspected mechanism. In principle, TK1 could exert its protective effects through supply of a supplementary dTTP pool for accurate repair of damaged genes; but Raji TK1(+) cells in thymidine free media still show preferential repair of transcribed regions. TK1 therefore does not exert its protective effects through dTTP pools, but through another unidentified mechanism, which affects sensitivity to and mutagenicity by DNA damaging agents.

  16. FISH comets show that the salvage enzyme TK1 contributes to gene-specific DNA repair

    Directory of Open Access Journals (Sweden)

    Stephen eDownes

    2014-08-01

    Full Text Available Thymidine kinase 1 (TK1 is a salvage enzyme that phosphorylates thymidine, imported from surrounding fluids, to create dTMP, which is further phosphorylated to the DNA precursor dTTP. TK1 deficiency has for a long time been known to cause increased cellular sensitivity to DNA damage. We have examined preferential strand break repair of DNA domains in TK1+ and TK1- clones of the Raji cell line, by the Comet-FISH technique, in bulk DNA and in the actively transcribed tumour suppressor (TP53 and human telomerase reverse transcriptase (hTERT gene regions, over 1 hour after 5Gy γ-irradiation. Results showed that repair of the TP53 and hTERT gene regions was more efficient in TK1+ compared to TK1- cells, while levels of genomic DNA repair were consistant between the two cell-lines. The targeted gene-specific repair in TK+ cells occurred rapidly, mainly over the first 15 minute repair-period. Therefore, TK1 is needed for preferential repair of actively transcribed regions, through a previously unsuspected mechanism. In principle, TK1 could exert its protective effects through supply of a supplementary dTTP pool for accurate repair of damaged genes; but Raji TK1+ cells in thymidine free media still show preferential repair of transcribed regions. TK1 therefore does not exert its protective effects through dTTP pools, but through another unidentified mechanism, which affects sensitivity to and mutagenicity by DNA damaging agents.

  17. N-tuple data viewer using Tcl/Tk

    International Nuclear Information System (INIS)

    Kubota, Toshimiti; Yoshida, Hajime

    1995-01-01

    Nowadays, UNIX/C users are minority in high energy physics community, while majority use VMS or mainframe FORTRAN programs. During five years in which efforts were exerted to SSC computing, the environment of HEP computing has drastically changed, and distributed computing and downsizing changed computing world. The strategy is to survey computing environment as widely as possible and to postpone the final choice and decision as late as possible. But some strategic decisions were done when necessary, such as the choice of UNIX and TCP/IP-based network system for the common working platform. The SSC computing activities to which the authors contributed are enumerated. It is important to evaluate the latest tool available for GUI, i.e., Tcl/Tk by implementing a practical tool, a n-tuple viewer. AVS is one of the most advanced scientific visualization tools, and the evaluation of the AVS for histogramming n-tuple data is discussed. The goal for the n-tuple viewer is shown. The design and the implementation of the n-tuple viewer for the evaluation of the Tcl/Tk as an interface builder are explained. The lessons and works to be done are pointed out. (K.I.)

  18. Quantification of thymidine kinase (TK1) mRNA in normal and leukemic cells and investigation of structure-function relatiosnhip of recombinant TK1enzyme

    DEFF Research Database (Denmark)

    Kristensen, Tina

    Thymidine kinase (TK) catalyses the ATP-dependent phosphorylation of thymidine to thymidine monophosphate, which is subsequency phosphorylated to thymidine triphosphate and utilized for DNA synthesis. Human cytosolic TK (TKI) is cell cycle regulated, e.g. the TK1 activity increases sharply at the G...... patients with chronic lymphatic leukemia (CLL). 2: Structure-function relationship of recombinant TKI. In the first part a sensitive method (competitive PCR) for quantification of TKI mRNA was established. The TKI mRNA level was quantified in quiescent lymphocytes from control donors (n = 6...... are characterized as being quiescent, the TK activity was in the same range as in quiescent lymphocytes from control donors. However, quantification of the TKI mRNA level shows that all five CLL patients had a very high level (6 to 22 x IO6 copies mg-’ protein) of TKI mRNA, corresponding to the level in dividing...

  19. Pengelolaan Kurikulum Sekolah Alam di TK Alam Al Biruni Cirebon

    Directory of Open Access Journals (Sweden)

    Ilma Fitriya Hidayati

    2016-04-01

    Full Text Available Tujuan penelitian ini adalahmendeskripsikan dan menganalisis: konsep, perencanaan, implementasi, dan evaluasi kurikulum sekolah alam di TK Alam Al Biruni Cirebon.Metode penelitian yang digunakan adalah metode kualitatif deskriptif dengan pendekatan studi kasus. Teknik pengumpulan data yang digunakan: wawancara, observasi, dan dokumentasi terhadap direktur pendidikan, kepala sekolah, guru, dan orang tua siswa. Sedangkan analisis data menggunakan prosedur: pengumpulan data, reduksi data, penyajian data, dan verifikasi data. Hasil penelitian menunjukkan: (1 kurikulum yang diterapkan merupakan perpaduan dari kurikulum nasional dan khas sekolah alam berbasis multiple intellegence berpilar pada akhlak serta terintegrasi menggunakan metode spider web, (2 perencanaan kurikulumdiwujudkan dalam bentuk silabus dan Rencana Kerja Harian; (3 pelaksanaan kurikulummenitik beratkan kepada pembentukan karakter dan akhlak; (4 evaluasi dilaksanakan berupa evaluasi mingguan, bulanan, dan tahunan.

  20. Aplikasi Media Pembelajaran Tematik untuk Anak TK Berbasis Android

    Directory of Open Access Journals (Sweden)

    Y. Haryo Sulistyanto Sunaryo

    2015-08-01

    Full Text Available Proses pembelajaran anak usia dini di Taman Kanak-kanak telah menggunakan pendekatan tematik dimana guru menggunakan beberapa tema sebagai fokus dari kegiatan pembelajaran. Tema-tema belajar yang disajikan dalam kegiatan pembelajaran diambil dari lingkungan kehidupan sekitar anak untuk mempermudah memberikan pembelajaran pada anak. Android merupakan sebuah sistem operasi berbasis kernel Linux. Android telah menjadi mobile platform paling populer di dunia. Aplikasi media pembelajaran Cerdas Tema dikembangkan sebagai sarana untuk memudahkan kegiatan pembelajaran siswa TK. Aplikasi Cerdas Tema diimplementasikan pada platform Android menggunakan Bahasa Lua dan terdiri dari beberapa tema diantaranya anggota tubuh, rumahku, binatang, dan tema bagian tanaman. Aplikasi Cerdas Tema memberikan pembelajaran tematik yang disajikan dalam bentuk gambar, suara, dan ejaan yang berdasarkan hasil kuesioner menunjukkan hasil persentase yang dominan baik atau mudah dimengerti oleh user dalam aspek grafis visual serta aspek entertainment dan pembelajaran. Kata Kunci: Aplikasi, Media Pembelajaran, Cerdas Tema, Pembelajaran Tematik, Taman Kanak-kanak

  1. Rapid application development using the Tcl/Tk language

    International Nuclear Information System (INIS)

    van Zeijts, J.

    1995-01-01

    During the last year, high level applications at CEBAF were written using the Tcl/Tk scripting language. This language is rapidly gaining in popularity, in part due to ease of constructing programs with X11 graphical user interfaces, and in part to ease of adding compiled user code for specialized purposes. Extensions to the language provide object oriented programming, which was used to develop a hierarchy of classes relevant for high level accelerator control. We describe basic language features, some 3rd party add-on packages, and local additions to the toolbox. Next we describe features of the accelerator object hierarchy, and finally describe applications written using this toolbox such as the ModelServer prototype, Slow Orbit and Energy Lock, the Linac Energy Management System, and other applications

  2. Dual monitoring using 124I-FIAU and bioluminescence for HSV1-tk suicide gene therapy

    International Nuclear Information System (INIS)

    Lee, T. S.; Kim, J. H.; Kwon, H. C.

    2007-01-01

    Herpes simplex virus type I thymidine kinase (HSV-tk) is the most common reporter gene and is used in cancer gene therapy with a prodrug nucleoside analog, ganciclovir (GCV). The aim of this study is to evaluate therapeutic efficacy of suicide gene therapy with 2'-fluoro-2'-deoxy-1-D-arabinofuranosyl-5-[ 124 I] iodouracil ( 124 I - FIAU) and bioluminescence in retrovirally HSV -tk and firefly luciferase transduced hepatoma model. The HSV -tk and firefly luciferase (Luc) was retrovirally transduced and expressed in MCA rat Morris hepatoma cells. Nude mice with subcutaneous tumors, MCA and MCA-TK-Luc, were subjected to GCV treatment (50mg/Kg/d intraperitoneally) for 5 day. PET imaging and biodistribution with ( 124 I-FIAU) were performed at before and after initiation of therapy with GCV. Bioluminescent signal was also measured during GCV treatment. Before GCV treatment, no significant difference in tumor volume was found in tumors between MCA and MCA-TK-Luc. After GCV treatment, tumor volume of MCA-TK-Luc markedly reduced compared to that of MCA. In biodistribution study, 124 I-FIAU uptake after GCV therapy significantly decreased compared with pretreatment levels (34.8 13.67 %ID/g vs 7.6 2.59 %ID/g) and bioluminescent signal was also significantly decreased compared with pretreatment levels. In small animal PET imaging, 124 I-FIAU selectively localized in HSV -tk expressing tumor and the therapeutic efficacy of GCV treatment was evaluated by 124 I-FIAU PET imaging. 124 I-FIAU PET and bioluminescence imaging in HSV-tk suicide gene therapy were effective to evaluate the therapeutic response. 124 I-FIAU may serve as an efficient and selective agent for monitoring of transduced HSV1-tk gene expression in vivo in clinical trials

  3. Geological Mapping of Investigation Trenches OL-TK15 and OL-TK16 at the Olkiluoto Study Site, Eurajoki, SW Finland

    International Nuclear Information System (INIS)

    Vaarma, M.; Vuokko, J.

    2009-07-01

    Geological mapping of investigation trenches OL-TK15 and OL-TK16 was carried out by the Geological Survey of Finland (GTK) at the Olkiluoto study site as a part of Posiva Oy's site investigation program for the development of an underground repository for nuclear waste. OL-TK15 is ca. N-S striking and ca. 95 m long, and OL-TK16 is ca. E-W striking and ca. 172 m long. The trenches were cleaned with pressure washer and pressurized air. The rock types were determined in field by naked eyes. Five samples from OL-TK15 and 10 samples from OL-TK16 were thin sectioned and investigated microscopically. In addition, petrophysical measurements were carried out by GTK geophysical laboratory for these samples. The bedrock within the excavation trenches OL-TK15 and OL-TK16 consists mainly of veined gneiss (VGN) with intercalations of mica gneiss (MGN), and mafic gneiss (MFGN) of amphibolite, amphibolite-gneiss, and skarn gneiss. The VGN is frankly solid or intact, fine grained and weakly banded and multiple intruded by granitic and pegmatitic veins and veinlets. Narrow scrappy zones occupied by dark dots of highly altered pseudomorphs after cordierite and/or garnet tend to be common in places. In addition, VGN contains rounded oblong mafic to intermediate fragments due to boudination of competent layers and/or dykes. Some rather narrow pegmatitic and quartz veins cut the previous tectonic structures. The prevailing tectonic structures are strong pervasive foliation and conformal veining of several generations mostly granitic in composition. The strike of the foliation and banding and veining as well, varies a little from NNE to ENE and the dip is ca. 40 - 50 degrees to SE sector, respectively. The younger folding has bent the older foliation and neosomic veining mostly with dextral monoclinic style so that in many places there occur tight kinky nodes and rootles augen like knots showing dextral rotation, too. Due to brittle deformation, there are some shear zones and zones with

  4. Comparative evaluation of TK SLC-L, a rapid liquid mycobacterial culture medium, with the MGIT system.

    Science.gov (United States)

    Çiftci, İhsan Hakkı; Karakeçe, Engin

    2014-03-06

    The present study was conducted to assess the efficiency of using TK SLC-L (Salubris, Inc.) for the primary isolation of mycobacteria from clinical samples by comparing it to the MGIT detection system (Becton Dickinson Diagnostic Instrument Systems). Although TK SLC, a biphasic medium, has been evaluated previously, this is the first study to evaluate TK SLC-L, a liquid medium. Clinical specimens from a total of 146 clinically suspected cases of tuberculosis were studied. Each processed sample was evaluated by ZN staining and inoculated into TK SLC-L and MGIT tubes. The TK SLC tubes were incubated in a MYCOLOR TK while the MGIT tubes were incubated in a MGIT system. Growth, indicated by automated systems, was confirmed through production of a smear and microscopic evaluation after ZN staining. Mycobacterial growth was positive in 35 TK SLC-L and in 34 MGIT samples. Although the growth detection time was approximately 3 to 5 days shorter, on average, with the MGIT system, the contamination rate was significantly lower using TK SLC-L. The total time spent for the repetition of cultures for contaminated samples in MGIT make the total return time for culture results equal to or longer than the time required by TK SLC-L. The TK Culture System using TK SLC-L is an efficient system and possible alternative to other rapid mycobacterial culture systems.

  5. Genetics Home Reference: TK2-related mitochondrial DNA depletion syndrome, myopathic form

    Science.gov (United States)

    ... American Mitochondrial Disease Consortium Resource List from the University of Kansas Medical Center: Mitochondrial ... Blakely E, He L, Gardner JL, Hudson G, Walter J, Hughes I, Turnbull DM, Taylor RW. Novel mutations in the TK2 gene associated ...

  6. Biological Education of IVFRU and FIAU for HSV1-TK Reporter Gene Monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Su Hee; Kim, Eun Jung; Lee, Eun Ah; Lee, Jong Chan; Choi, Tae Hyun; Lee, Kyo Chul; An, Gwang Il; Cheon, Gi Jeong [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2006-07-01

    The Herpes Simplex Virus Type1-thymidine kinase (HSV1-TK) system is a useful gene therapy monitoring method. HSV1-TK is one of the most widely used effector gene systems used for imaging gene expression, in association with its use as a reporter gene. It has resulted the development of a number of radiolabeled HSV1-TK substrates for the non-invasive detection of HSV1-TK expression. In non-invasive imaging of the HSV1-TK system, many nucleoside derivatives have been developed as prodrugs for tumor proliferation imaging or as anti-viral drugs. Prodrug activation or sucide gene therapy has been shown to be successful in potentiating the therapeutic index by sensitizing genetically modified tumor cells to various prodrugs or enhancing the action of commonly used chemotherapeutic agents. The most studied prodrug activation approaches involve transfection of tumors with HSV1-TK gene. (Z)-5-(2-iodovinyl)-2'-fluoro- 2'-deoxyuridine (IVFRU) possesses a 2'-fluoro substituent in the ribose configuration, is considered to protect IVFRU from enzyme mediated degradation in vivo. It is obviously potential substrates for HSV1-TK imaging. 2'-Fiuoro-2'-deoxy-1-{beta}-D-arabinofuranosyl- 5-iodo-uridine (FIAU), an anticancer drug widely used in clinical practice, is an analogue of thymidine. In a series of studies using adenovirus vector for gene transfer described the appropriate combination of exogenously introduced HSV1-TK as a 'marker/reporter gene' and radiolabelled FIAU as a 'marker substrate/reporter probe' for monitoring gene therapy and gene expression.

  7. Biological Education of IVFRU and FIAU for HSV1-TK Reporter Gene Monitoring

    International Nuclear Information System (INIS)

    Hong, Su Hee; Kim, Eun Jung; Lee, Eun Ah; Lee, Jong Chan; Choi, Tae Hyun; Lee, Kyo Chul; An, Gwang Il; Cheon, Gi Jeong

    2006-01-01

    The Herpes Simplex Virus Type1-thymidine kinase (HSV1-TK) system is a useful gene therapy monitoring method. HSV1-TK is one of the most widely used effector gene systems used for imaging gene expression, in association with its use as a reporter gene. It has resulted the development of a number of radiolabeled HSV1-TK substrates for the non-invasive detection of HSV1-TK expression. In non-invasive imaging of the HSV1-TK system, many nucleoside derivatives have been developed as prodrugs for tumor proliferation imaging or as anti-viral drugs. Prodrug activation or sucide gene therapy has been shown to be successful in potentiating the therapeutic index by sensitizing genetically modified tumor cells to various prodrugs or enhancing the action of commonly used chemotherapeutic agents. The most studied prodrug activation approaches involve transfection of tumors with HSV1-TK gene. (Z)-5-(2-iodovinyl)-2'-fluoro- 2'-deoxyuridine (IVFRU) possesses a 2'-fluoro substituent in the ribose configuration, is considered to protect IVFRU from enzyme mediated degradation in vivo. It is obviously potential substrates for HSV1-TK imaging. 2'-Fiuoro-2'-deoxy-1-β-D-arabinofuranosyl- 5-iodo-uridine (FIAU), an anticancer drug widely used in clinical practice, is an analogue of thymidine. In a series of studies using adenovirus vector for gene transfer described the appropriate combination of exogenously introduced HSV1-TK as a 'marker/reporter gene' and radiolabelled FIAU as a 'marker substrate/reporter probe' for monitoring gene therapy and gene expression

  8. Characterization of a TK6-Bcl-xL gly-159-ala Human Lymphoblast Clone

    Energy Technology Data Exchange (ETDEWEB)

    Chyall, L.: Gauny, S.; Kronenberg, A.

    2006-01-01

    TK6 cells are a well-characterized human B-lymphoblast cell line derived from WIL-2 cells. A derivative of the TK6 cell line that was stably transfected to express a mutated form of the anti-apoptotic protein Bcl-xL (TK6-Bcl-xL gly-159- ala clone #38) is compared with the parent cell line. Four parameters were evaluated for each cell line: growth under normal conditions, plating efficiency, and frequency of spontaneous mutation to 6‑thioguanine resistance (hypoxanthine phosphoribosyl transferase locus) or trifluorothymidine resistance (thymidine kinase locus). We conclude that the mutated Bcl-xL protein did not affect growth under normal conditions, plating efficiency or spontaneous mutation frequencies at the thymidine kinase (TK) locus. Results at the hypoxanthine phosphoribosyl transferase (HPRT) locus were inconclusive. A mutant fraction for TK6‑Bcl-xL gly-159-ala clone #38 cells exposed to 150cGy of 160kVp x-rays was also calculated. Exposure to x-irradiation increased the mutant fraction of TK6‑Bcl-xL gly-159-ala clone #38 cells.

  9. [Growing characteristic of breast adenocarcinoma suicide gene cell line T47D-tk and construction of subcutaneous xenografts].

    Science.gov (United States)

    Xu, Wen-Gui; Yang, Kun; Fang, Na; Dai, Dong; Yu, Jin-Pu; Wei, Feng; Song, Xiu-Yu; Zhu, Yan-Jia; Wang, Jian; Men, Xiao-Yuan; Zhang, Ying

    2009-02-17

    To further identify the breast adenocarcinoma cell line T47D-tk stably expressing the suicide gene HSV1-tk, observe its growing characteristics, and establish an animal model of implanted breast adenocarcinoma. Retroviral vector of HSV1-tk gene and breast carcinoma cell line T47D-tk stably expressing the HSV1-tk gene were established. Breast carcinoma cells of the lines T47D and T47D-tk were cultured, and observed by inverted microscope. Growth curve was drawn. Genomic DNA of T47D-tk cells was extracted, and amplified by PCR with HSV1-tk and HSV1-tk P2 as primers. Agarose gel electrophoresis was used to identify the HSV1tk gene. Suspensions of T47D and T47D-tk cells were inoculated subcutaneously at bilateral roots of foreleg of female BALB/c nude mice respectively. The growth of tumor was observed every day. PCR showed 1131 bp fragment in the T47D-tk genome, but not in the T47D genome. The numbers of growing T47D cells on days 3, and 7 were (10.00 +/- 1.30) x 10(3) and (19.25 +/- 0.66) x 10(3) respectively, not significantly different from those of the T47D-tk cells [(10.25 +/- 0.90) x 10(3) and (19.00 +/- 1.80) x 10(3) respectively, both P > 0.05]. The time needed for tumor formation after the inoculation of T47D cells was (9.67 +/- 0.33) d, not significantly different from that of the T47D-tk cells [(9.83 +/- 0.48) d, P > 0.05]. The tumor size 19 days after inoculation of the T47D cells was (72.17 +/- 25.88) mm(3), not significantly different from that of the T47D-tk cells [(70.66 +/- 22.16) mm(3), P > 0.05]. The T47D-tk cells have integrated the HSV1-tk gene without changing its growing characteristics. An animal model of implanted breast cancer has been successfully established. The T47D and T47D-tk subcutaneous xenografts offer the foundation for further study of gene imaging and gene therapy.

  10. Dexamethasone inhibits the HSV-tk/ ganciclovir bystander effect in malignant glioma cells

    Directory of Open Access Journals (Sweden)

    Jolois Olivier

    2005-04-01

    Full Text Available Abstract Background HSV-tk/ ganciclovir (GCV gene therapy has been extensively studied in the setting of brain tumors and largely relies on the bystander effect. Large studies have however failed to demonstrate any significant benefit of this strategy in the treatment of human brain tumors. Since dexamethasone is a frequently used symptomatic treatment for malignant gliomas, its interaction with the bystander effect and the overall efficacy of HSV-TK gene therapy ought to be assessed. Methods Stable clones of TK-expressing U87, C6 and LN18 cells were generated and their bystander effect on wild type cells was assessed. The effects of dexamethasone on cell proliferation and sensitivity to ganciclovir were assessed with a thymidine incorporation assay and a MTT test. Gap junction mediated intercellular communication was assessed with microinjections and FACS analysis of calcein transfer. The effect of dexamethasone treatment on the sensitivity of TK-expressing to FAS-dependent apoptosis in the presence or absence of ganciclovir was assessed with an MTT test. Western blot was used to evidence the effect of dexamethasone on the expression of Cx43, CD95, CIAP2 and BclXL. Results Dexamethasone significantly reduced the bystander effect in TK-expressing C6, LN18 and U87 cells. This inhibition results from a reduction of the gap junction mediated intercellular communication of these cells (GJIC, from an inhibition of their growth and thymidine incorporation and from a modulation of the apoptotic cascade. Conclusion The overall efficacy of HSV-TK gene therapy is adversely affected by dexamethasone co-treatment in vitro. Future HSV-tk/ GCV gene therapy clinical protocols for gliomas should address this interference of corticosteroid treatment.

  11. Imaging expression of adenoviral HSV1-tk suicide gene transfer using the nucleoside analogue FIRU

    International Nuclear Information System (INIS)

    Nanda, Dharmin; Jong, Marion de; Bakker, Willem; Bijster, Magda; Cox, Peter; Vogels, Ronald; Havenga, Menzo; Driesse, Maarten; Avezaat, Cees; Morin, Kevin; Naimi, Ebrahim; Knaus, Edward; Wiebe, Leonard; Smitt, Peter Sillevis

    2002-01-01

    Substrates for monitoring HSV1-tk gene expression include uracil and acycloguanosine derivatives.The most commonly used uracil derivative to monitor HSV1-tk gene transfer is 1-(2-fluoro-2-deoxy-β-D-arabinofuranosyl)-5-[*I]iodouracil (fialuridine; I*-FIAU), where the asterisk denotes any of the radioactive iodine isotopes that can be used. We have previously studied other nucleosides with imaging properties as good as or better than FIAU, including 1-(2-fluoro-2-deoxy-β-D-ribofuranosyl)-5-[*I]iodouracil (FIRU). The first aim of this study was to extend the biodistribution data of 123 I-labelled FIRU. Secondly, we assessed the feasibility of detecting differences in HSV1-tk gene expression levels following adenoviral gene transfer in vivo with 123 I-FIRU. 9L rat gliosarcoma cells were stably transfected with the HSV1-tk gene (9L-tk+). 123 I-FIRU was prepared by radioiodination of 1-(2-fluoro-2-deoxy-β-D-ribofuranosyl)-5-tributylstannyl uracil (FTMRSU; precursor compound) and purified using an activated Sep-Pak column. Incubation of 9L-tk+ cells and the parental 9L cells with 123 I-FIRU resulted in a 100-fold higher accumulation of radioactivity in the 9L-tk+ cells after an optimum incubation time of 4 h. NIH-bg-nu-xid mice were then inoculated subcutaneously with HSV1-tk (-) 9L cells or HSV1-tk (+) 9L-tk+ cells into both flanks. Biodistribution studies and gamma camera imaging were performed at 15 min and 1, 2, 4 and 24 h p.i. At 15 min, the tumour/muscle, tumour/blood and tumour/brain ratios were 5.2, 1.0 and 30.3 respectively. Rapid renal clearance of the tracer from the body resulted in increasing tumour/muscle, tumour/blood and tumour/brain ratios, reaching values of 32.2, 12.5 and 171.6 at 4 h p.i. A maximum specific activity of 22%ID/g tissue was reached in the 9L-tk+ tumours 4 h after 123 I-FIRU injection. Two Ad5-based adenoviral vectors containing the HSV1-tk gene were constructed: a replication-incompetent vector with the transgene in the former E1

  12. Sampling and analysis plan (SAP) for WESF drains and TK-100 sump

    International Nuclear Information System (INIS)

    Simmons, F.M.

    1998-01-01

    The intent of this project is to determine whether the Waste Encapsulation and Storage Facility (WESF) floor drain piping and the TK-100 sump are free from contamination. TK-100 is currently used as a catch tank to transfer low level liquid waste from WESF to Tank Farms via B Plant. This system is being modified as part of the WESF decoupling since B Plant is being deactivated. As a result of the 1,1,1-trichloroethane (TCA) discovery in TK-100, the associated WESF floor drains and the pit sump need to be sampled. Breakdown constituents have been reviewed and found to be non-hazardous. There are 29 floor drains that tie into a common header leading into the tank. To prevent high exposure during sampling of the drains, TK-100 will be removed into the B Plant canyon and a new tank will be placed in the pit before any floor drain samples are taken. The sump will be sampled prior to TK-100 removal. A sample of the sludge and any liquid in the sump will be taken and analyzed for TCA and polychlorinated biphenyl (PCB). After the sump has been sampled, the vault floor will be flushed. The flush will be transferred from the sump into TK-100. TK-100 will be moved into B Plant. The vault will then be cleaned of debris and visually inspected. If there is no visual indication of TCA or PCB staining, the vault will be painted and a new tank installed. If there is an indication of TCA or PCB from laboratory analysis or staining, further negotiations will be required to determine a path forward. A total of 8 sets of three 40ml samples will be required for all of the floor drains and sump. The sump set will include one 125ml solid sample. The only analysis required will be for TCA in liquids. PCBs will be checked in sump solids only. The Sampling and Analysis Plan (SAP) is written to provide direction for the sampling and analytical activities of the 29 WESF floor drains and the TK-100 sump. The intent of this plan is to define the responsibilities of the various organizations

  13. Data Analysis of the TK-1G Sounding Rocket Installed with a Satellite Navigation System

    Directory of Open Access Journals (Sweden)

    Lesong Zhou

    2017-10-01

    Full Text Available This article gives an in-depth analysis of the experimental data of the TK-1G sounding rocket installed with the satellite navigation system. It turns out that the data acquisition rate of the rocket sonde is high, making the collection of complete trajectory and meteorological data possible. By comparing the rocket sonde measurements with those obtained by virtue of other methods, we find that the rocket sonde can be relatively precise in measuring atmospheric parameters within the scope of 20–60 km above the ground. This establishes the fact that the TK-1G sounding rocket system is effective in detecting near-space atmospheric environment.

  14. TRAIL causes deletions at the HPRT and TK1 loci of clonogenically competent cells

    Energy Technology Data Exchange (ETDEWEB)

    Miles, Mark A.; Shekhar, Tanmay M. [Department of Biochemistry and Genetics, La Trobe University, Bundoora, Victoria (Australia); La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria (Australia); Hall, Nathan E. [La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria (Australia); Life Sciences Computation Centre, Victorian Life Sciences Computation Initiative, Melbourne, Victoria (Australia); Hawkins, Christine J., E-mail: c.hawkins@latrobe.edu.au [Department of Biochemistry and Genetics, La Trobe University, Bundoora, Victoria (Australia); La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria (Australia)

    2016-05-15

    Highlights: • Treatment with TRAIL or EMS provokes mutations in clonogenically viable TK6 cells. • TRAIL is 2–5-fold less mutagenic than an equivalently lethal concentration of EMS. • EMS mainly causes transition mutations at the HPRT and TK1 loci of TK6 cells. • Most loss-of-function HPRT or TK1 mutations caused by TRAIL treatment are deletions. - Abstract: When chemotherapy and radiotherapy are effective, they function by inducing DNA damage in cancerous cells, which respond by undergoing apoptosis. Some adverse effects can result from collateral destruction of non-cancerous cells, via the same mechanism. Therapy-related cancers, a particularly serious adverse effect of anti-cancer treatments, develop due to oncogenic mutations created in non-cancerous cells by the DNA damaging therapies used to eliminate the original cancer. Physiologically achievable concentrations of direct apoptosis inducing anti-cancer drugs that target Bcl-2 and IAP proteins possess negligible mutagenic activity, however death receptor agonists like TRAIL/Apo2L can provoke mutations in surviving cells, probably via caspase-mediated activation of the nuclease CAD. In this study we compared the types of mutations sustained in the HPRT and TK1 loci of clonogenically competent cells following treatment with TRAIL or the alkylating agent ethyl methanesulfonate (EMS). As expected, the loss-of-function mutations in the HPRT or TK1 loci triggered by exposure to EMS were almost all transitions. In contrast, only a minority of the mutations identified in TRAIL-treated clones lacking HPRT or TK1 activity were substitutions. Almost three quarters of the TRAIL-induced mutations were partial or complete deletions of the HPRT or TK1 genes, consistent with sub-lethal TRAIL treatment provoking double strand breaks, which may be mis-repaired by non-homologous end joining (NHEJ). Mis-repair of double-strand breaks following exposure to chemotherapy drugs has been implicated in the pathogenesis of

  15. ClearTK 2.0: Design Patterns for Machine Learning in UIMA.

    Science.gov (United States)

    Bethard, Steven; Ogren, Philip; Becker, Lee

    2014-05-01

    ClearTK adds machine learning functionality to the UIMA framework, providing wrappers to popular machine learning libraries, a rich feature extraction library that works across different classifiers, and utilities for applying and evaluating machine learning models. Since its inception in 2008, ClearTK has evolved in response to feedback from developers and the community. This evolution has followed a number of important design principles including: conceptually simple annotator interfaces, readable pipeline descriptions, minimal collection readers, type system agnostic code, modules organized for ease of import, and assisting user comprehension of the complex UIMA framework.

  16. Autoradiography study and SPECT imaging of reporter gene HSV1-tk expression in heart

    Energy Technology Data Exchange (ETDEWEB)

    Lan Xiaoli [Department of Nuclear Medicine, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, Hubei Province, 430022 (China)], E-mail: LXL730724@hotmail.com; Liu Ying; He Yong; Wu Tao; Zhang Binqing; Gao Zairong; An Rui [Department of Nuclear Medicine, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, Hubei Province, 430022 (China); Zhang Yongxue [Department of Nuclear Medicine, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, Hubei Province, 430022 (China)], E-mail: zhyx1229@163.com

    2010-04-15

    Aim: To demonstrate the feasibility and optimal conditions of imaging herpes simplex virus 1-thymidine kinase (HSV1-tk) gene transferred into hearts with {sup 131}I-2'-fluoro-2'-deoxy-1-{beta}-D-arabinofuranosyl-5-iodouracil ({sup 131}I-FIAU) using autoradiography (ARG) and single photon emission computed tomography (SPECT) in animal models. Methods: HSV1-tk inserted into adenovirus vector (Ad5-tk) and adenovirus (Ad5-null) was prepared. Rats or rabbits were divided into a study group receiving intramyocardial injection of Ad5-tk, and a control group receiving Ad-null injection. In the study group of rats, two sets of experiments, time-course study and dose-dependence study, were performed. In time-course experiments, rats were injected with {sup 131}I-FIAU on Days 1, 2, 3, 5 and 7, after transfection of 1x10{sup 8} pfu Ad5-tk, to study the feasibility and suitable time course for reporter gene imaging. In dose-dependence study, various titers of Ad5-tk (5x10{sup 8}, 1x10{sup 8}, 5x10{sup 7} and 1x10{sup 7} pfu) were used to determine the threshold and optimal viral titer needed for detection of gene expression. The gamma counts of hearts were measured. The rat myocardium was analyzed by ARG and reverse transcriptase-polymerase chain reaction (RT-PCR). SPECT whole-body planar imaging and cardiac tomographic imaging were performed in the rabbit models. Results: From the ARG images, rats injected with Ad5-tk showed significant {sup 131}I-FIAU activity in the anterolateral wall compared with background signals seen in the control Ad5-null rats. In time-course study, the highest radioactivity in the focal myocardium could be seen on Day 1, and then progressively declined with time. In dose-dependence study, the level of {sup 131}I-FIAU accumulation in the transfected myocardium declined with the decrease of Ad viral titers. From the ARG analysis and gamma counting, the threshold viral titer was 5x10{sup 7} pfu, and the optimal Ad titer was 1x10{sup 8} pfu

  17. Real-time dynamic optical imaging of ACC-M tumor cells killed by HSV-tk/ACV system.

    Science.gov (United States)

    Xiong, Tao; Li, Yongjin; Li, Zhiyang; Xie, Xiangmo; Lu, Lisha

    2013-01-01

    HSV-tk/ACV induced and killed human adenoid cystic carcinoma cell (ACC-M) in vivo and in vitro, which were observed through optical imaging and green fluorescence protein (GFP) tagging technique. ACC-M was transfected with TK-GFP, and the single clone cell ACC-M-TK-GFP was selected by G418. With fluorescent stereomicroscope, whole-body fluorescent imaging system and fluorescent microscope, we could observe ACV treated ACC-M-TK-GFP cells in cell level and nude mice. The therapies of tumor were visualized clearly with optical imaging. This study proves that optical imaging is a very good approach for studying the effect of HSV-tk/ACV on the ACC-M tumor cells and decreasing the amount of vessel about tumors cell. Optical imaging will become a visual groundwork for monitoring tumor growth and evaluating in vivo curative effect of antitumor drugs.

  18. MerTK Cleavage on Resident Cardiac Macrophages Compromises Repair After Myocardial Ischemia Reperfusion Injury.

    Science.gov (United States)

    DeBerge, Matthew; Yeap, Xin Yi; Dehn, Shirley; Zhang, Shuang; Grigoryeva, Lubov; Misener, Sol; Procissi, Daniel; Zhou, Xin; Lee, Daniel C; Muller, William A; Luo, Xunrong; Rothlin, Carla; Tabas, Ira; Thorp, Edward B

    2017-09-29

    Clinical benefits of reperfusion after myocardial infarction are offset by maladaptive innate immune cell function, and therapeutic interventions are lacking. We sought to test the significance of phagocytic clearance by resident and recruited phagocytes after myocardial ischemia reperfusion. In humans, we discovered that clinical reperfusion after myocardial infarction led to significant elevation of the soluble form of MerTK (myeloid-epithelial-reproductive tyrosine kinase; ie, soluble MER), a critical biomarker of compromised phagocytosis by innate macrophages. In reperfused mice, macrophage Mertk deficiency led to decreased cardiac wound debridement, increased infarct size, and depressed cardiac function, newly implicating MerTK in cardiac repair after myocardial ischemia reperfusion. More notably, Mertk(CR ) mice, which are resistant to cleavage, showed significantly reduced infarct sizes and improved systolic function. In contrast to other cardiac phagocyte subsets, resident cardiac MHCII LO CCR2 - (major histocompatibility complex II/C-C motif chemokine receptor type 2) macrophages expressed higher levels of MerTK and, when exposed to apoptotic cells, secreted proreparative cytokines, including transforming growth factor-β. Mertk deficiency compromised the accumulation of MHCII LO phagocytes, and this was rescued in Mertk(CR ) mice. Interestingly, blockade of CCR2-dependent monocyte infiltration into the heart reduced soluble MER levels post-ischemia reperfusion. Our data implicate monocyte-induced MerTK cleavage on proreparative MHCII LO cardiac macrophages as a novel contributor and therapeutic target of reperfusion injury. © 2017 American Heart Association, Inc.

  19. A philosophical assessment of TK's autopsy report: Implications for the debate over the brain death criteria.

    Science.gov (United States)

    Austriaco, Nicanor Pier Giorgio

    2016-05-01

    In recent years, there has been increasing evidence that the totally brain-dead patient is able to continue to live and to maintain some integrated functions, albeit with the necessary assistance of mechanical ventilation. Several years ago, the autopsy report of a totally brain-dead patient named TK who was kept on life support for nearly twenty years was published in the Journal of Child Neurology. He remains the individual kept on life support the longest after suffering total brain failure. In this essay, I argue that the clinical data described in the autopsy report demonstrate that TK's long-term survival after total brain failure supports the claim acknowledged by the President's Council on Bioethics that the brain-dead patient retains his bodily integrity. As such, he is not dead. He is still a living, though severely disabled, human organism, a human person made in the image and likeness of God. Traditionally, the presence or absence of bodily integration has been used to definitively discern the presence or absence of life in the human being where decomposition of the body is the surest sign of death. The autopsy report of a patient named TK who was brain-dead for nearly twenty years demonstrates that brain-dead patients retain their bodily integrity. As such, TK and other brain-dead patients are not dead. They are living, though severely disabled, human organisms, who are human persons made in the image and likeness of God.

  20. HUBUNGAN ANTARA FLAT FOOT DENGAN KESEIMBANGAN DINAMIS PADA MURID TK SULAWESI KOTA MAKASSAR

    OpenAIRE

    SAHABUDDIN, HUMAIRAH

    2016-01-01

    2016 Humairah Sahabuddin, C13112002. ???Hubungan antara Flat Foot dengan Keseimbangan Dinamis pada Murid TK Sulawesi Kota Makassar???. Dibimbing oleh Muliyadi dan Nindrahayu. Latar belakang: Kemampuan keseimbangan anak pada usia perkembangan sangat penting untuk ditinjau, baik dari segi keseimbangan duduk, berdiri, dan saat berjalan. Salah satu faktor yang dapat menyebabkan gangguan keseimbangan pada anak yaitu gangguan muskuloskeletal berupa kelainan bentuk telapak kaki ...

  1. Decreased TK activity alters growth, yield and tolerance to low temperature and low light intensity in transgenic cucumber plants.

    Science.gov (United States)

    Bi, Huangai; Dong, Xubing; Wu, Guoxiu; Wang, Meiling; Ai, Xizhen

    2015-02-01

    Four CsTK antisense transgenic cucumber plants were obtained. Decreased TK activity decreased the photosynthetic rate, seed germination rate, growth yield, and the tolerance to low temperature and weak light stress. Transketolase (TK, EC 2.2.1.1) is a key enzyme in the photosynthetic carbon reduction cycle (Calvin cycle). A cDNA fragment (526 bp) encoding transketolase was cloned from cucumber plants (Cucumis sativa L. cv 'Jinyou 3') by RT-PCR. The antisense expression [(PBI-CsTK(-)] vector containing the CsTK gene fragment was constructed. The resulting plasmid was introduced into the cucumber inbred lines '08-1' using the agrobacterium-mediated method, and four antisense transgenic cucumber plants were obtained. Decreased CsTK expression either unaltered or slightly increased the mRNA abundance and activities of the other main enzymes in the Calvin cycle, however, it decreased the TK activity and net photosynthetic rate (Pn) in antisense transgenic cucumber leaves. Antisense plants showed decreases in the growth, ratio of female flowers and yield compared with the wild-type (WT) plants. The decrease in Pn, stomatal conductance (Gs), transpiration rate (Tr), photochemical efficiency (Fv/Fm) and actual photochemical efficiency of PSII (ΦPSII) and the increase in electrolyte leakage (EL) were greater in antisense transgenic plants than in WT plants under low temperature (5 °C) and low light intensity (100 μmol m(-2) s(-1)).

  2. Dual monitoring using {sup 124}I-FIAU and bioluminescence for HSV1-tk suicide gene therapy

    Energy Technology Data Exchange (ETDEWEB)

    Lee, T. S.; Kim, J. H.; Kwon, H. C. [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)] (and others)

    2007-07-01

    Herpes simplex virus type I thymidine kinase (HSV-tk) is the most common reporter gene and is used in cancer gene therapy with a prodrug nucleoside analog, ganciclovir (GCV). The aim of this study is to evaluate therapeutic efficacy of suicide gene therapy with 2'-fluoro-2'-deoxy-1-D-arabinofuranosyl-5-[{sup 124}I] iodouracil ({sup 124}I - FIAU) and bioluminescence in retrovirally HSV -tk and firefly luciferase transduced hepatoma model. The HSV -tk and firefly luciferase (Luc) was retrovirally transduced and expressed in MCA rat Morris hepatoma cells. Nude mice with subcutaneous tumors, MCA and MCA-TK-Luc, were subjected to GCV treatment (50mg/Kg/d intraperitoneally) for 5 day. PET imaging and biodistribution with ({sup 124}I-FIAU) were performed at before and after initiation of therapy with GCV. Bioluminescent signal was also measured during GCV treatment. Before GCV treatment, no significant difference in tumor volume was found in tumors between MCA and MCA-TK-Luc. After GCV treatment, tumor volume of MCA-TK-Luc markedly reduced compared to that of MCA. In biodistribution study, {sup 124}I-FIAU uptake after GCV therapy significantly decreased compared with pretreatment levels (34.8 13.67 %ID/g vs 7.6 2.59 %ID/g) and bioluminescent signal was also significantly decreased compared with pretreatment levels. In small animal PET imaging, {sup 124}I-FIAU selectively localized in HSV -tk expressing tumor and the therapeutic efficacy of GCV treatment was evaluated by {sup 124}I-FIAU PET imaging. {sup 124}I-FIAU PET and bioluminescence imaging in HSV-tk suicide gene therapy were effective to evaluate the therapeutic response. {sup 124}I-FIAU may serve as an efficient and selective agent for monitoring of transduced HSV1-tk gene expression in vivo in clinical trials.

  3. Synthesis and Cellular Uptake of Radioiodine labeled 2{sup '}-deoxyuridine derivatives with HSV1-TK

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Eun Ah; Lee, Kyo Chul; Hong, Su Hee; Kim, Eun Jung; Lee, Jong Chan; An, Gwang Il; Choi, Tae Hyun; Cheon, Gi Jeong; Chun, Kwon Soo [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2006-07-01

    Several different radiolabeled probes have been developed to image Herpes Simplex Virus Type-1 thyrimidine kinase gene (HSV1-TK) expression. The nucleoside prodrugs under investigation for HSV1-TK imaging generally fall into two main chemical and radioisotope categories: the pyrimidine nucleosides, primarily radioiodinated, and the purin nucleosides, primarily radiofluorinated, and their respective analogues. In non-invasive imaging of the HSV1-TK system, many nucleoside derivatives have been recommended as HSV1-TK substrates. Most of these nucleoside derivatives have been developed as prodrugs for tumor proliferation imaging or as anti-viral drugs. For example, 5-fluorouracil (5-FU) and IUdR have been used as tumor agents and acyclovir (ACV), ganciclovir (GCV) and (E)-5-(2-bromovinyl)-2{sup '}- deoxyuridine (BVDU) as an anti-viral agents for virus infection several 5-substituted uracil nucleoside derivatives have been identified to have high sensitivity and selective accumulation in HSV1-TK expressing cells. Of those, IVDU was shown to be rapidly accumulated in HSV1- TK expressing cells in vitro. Imaging of the HSV1-TK reporter gene along with various reporter probes is of current interest. In contrast to the mammalian kinase, which phosphorylates thymidine preferentially, HSV1-TK is less discriminative and phosphorylates a wide range of nucleoside analogues such as acycloguanosines and 2{sup '}-fluoro-2{sup '}-deoxyuridine derivatives that are not phosphorylated efficiently by the native enzyme. More specifically, 5-substituted 2{sup '}-fluoro-2{sup '}-deoxyarabinofuranosyluracil nucleosides are efficiently phosphorylated by HSV- TK. This property, together with the presence of fluorine in the 2{sup '}-arabino-position, endows the 2{sup '}-fluoro-2{sup '}-deoxyuridines with antiviral activity against HSV.

  4. MALDI MS

    Indian Academy of Sciences (India)

    Unknown

    Abstract. Matrix assisted laser desorption ionization mass spectroscopy (MALDI MS) is a technique which allows the measurement of molecular mass > 200,000 Daltons by ionization and vapourization without degra- dation. This technique is useful for the mass analysis of synthetic polymers, which have very low volatility.

  5. PET imaging of HSV1-tk mutants with acquired specificity toward pyrimidine- and acycloguanosine-based radiotracers

    Energy Technology Data Exchange (ETDEWEB)

    Likar, Yury; Dobrenkov, Konstantin; Olszewska, Malgorzata; Shenker, Larissa; Hricak, Hedvig; Ponomarev, Vladimir [Memorial Sloan-Kettering Cancer Center, Molecular Imaging Laboratory, Department of Radiology, New York, NY (United States); Cai, Shangde [Memorial Sloan-Kettering Cancer Center, Radiochemistry/Cyclotron Core Facility, New York, NY (United States)

    2009-08-15

    The aim of this study was to create an alternative mutant of the herpes simplex virus type 1 thymidine kinase (HSV1-tk) reporter gene with reduced phosphorylation capacity for acycloguanosine derivatives, but not pyrimidine-based compounds that will allow for successful PET imaging. A new mutant of HSV1-tk reporter gene, suitable for PET imaging using pyrimidine-based radiotracers, was developed. The HSV1-tk mutant contains an arginine-to-glutamine substitution at position 176 (HSV1-R176Qtk) of the nucleoside binding region of the enzyme. The mutant-gene product showed favorable enzymatic characteristics toward pyrimidine-based nucleosides, while exhibiting reduced activity with acycloguanosine derivatives. In order to enhance HSV1-R176Qtk reporter activity with pyrimidine-based radiotracers, we introduced the R176Q substitution into the more active HSV1-sr39tk mutant. U87 human glioma cells transduced with the HSV1-R176Qsr39tk double mutant reporter gene showed high {sup 3}H-FEAU pyrimidine nucleoside and low {sup 3}H-penciclovir acycloguanosine analog uptake in vitro. PET imaging also demonstrated high {sup 18}F-FEAU and low {sup 18}F-FHBG accumulation in HSV1-R176Qsr39tk+ xenografts. The feasibility of imaging two independent nucleoside-specific HSV1-tk mutants in the same animal with PET was demonstrated. Two opposite xenografts expressing the HSV1-R176Qsr39tk reporter gene and the previously described acycloguanosine-specific mutant of HSV1-tk, HSV1-A167Ysr39tk reporter gene, were imaged using a short-lived pyrimidine-based {sup 18}F-FEAU and an acycloguanosine-based {sup 18}F-FHBG radiotracer, respectively, administered on 2 consecutive days. We conclude that in combination with acycloguanosine-specific HSV1-A167Ysr39tk reporter gene, a HSV1-tk mutant containing the R176Q substitution could be used for PET imaging of two different cell populations or concurrent molecular biological processes in the same living subject. (orig.)

  6. Model of Religious Study and Moral Values in TK Putra Harapan Nalumsari Jepara

    Directory of Open Access Journals (Sweden)

    Mubasyaroh Mubasyaroh

    2016-12-01

    Full Text Available Religious and moral education from an early age so needs to be invested for the child, so that in the future they will have a strong and deep understanding of the norms and teachings of Islam. Age children early childhood and kindergarten (TK is a time to play, so education is implemented particularly religious education should be designed properly by the teacher so that the education process into an active and fun activities. Kindergarten (TK Putra Harapan Nalumsari Jepara is one of the educational institutions that provide education for early childhood, with one lesson material is a moral and religious education. Moral education is one of the materials is very important because in load values, morals and religion, so this will be a guide for students in later life. The research method used is descriptive qualitative research to describe in detail the subjects and issues to be studied. The findings in this study is the cultivation of religious values and morals for children Kindergarten revolves around the activities of daily life. In particular cultivation of religious values with laying the foundations of the faith, personality or character that is commendable and devotional practices, in accordance with the child's ability is implemented. As one of the early childhood education institutions, in conducting the study, have several models of delivery Edutainment, habituation and uswah hasanah as a reference implementation of teaching and learning. Pendidikan agama dan moral sejak usia dini perlu ditanamkan bagi anak, sehingga di masa depan mereka akan memiliki pemahaman yang kuat dan mendalam dari norma-norma dan ajaran Islam. Usia anak-anak usia dini dan taman kanak-kanak (TK adalah waktu untuk bermain, sehingga pendidikan diimplementasikan khususnya pendidikan agama harus dirancang dengan baik oleh guru sehingga proses pendidikan menjadi kegiatan yang aktif dan menyenangkan. TK (TK Putra Harapan Nalumsari Jepara merupakan salah satu lembaga

  7. Genotoxic and mutagenic potential of camphorquinone in L5178/TK+/-mouse lymphoma cells.

    Science.gov (United States)

    Volk, J; Ziemann, C; Leyhausen, G; Geurtsen, W

    2018-03-01

    Camphorquinone (CQ) is the most important photoinitiator used in dental composite resins. Sparse data indicate a mutagenic potential of CQ. Therefore, it was aim of this study to evaluate the cytotoxicity, genotoxicity, and mutagenicity of CQ in L5178Y TK +/- mouse lymphoma cells. L5178Y/TK +/- cells were exposed to different concentrations of non-irradiated CQ (0.25-2.5mM). Cytotoxicity was evaluated by propidium iodide assay, determination of suspension growth rate, relative total growth and the mitotic index. Intracellular levels of reactive oxygen/nitrogen species (ROS/RNS) were quantified by 2',7'-dichlorofluoresceine diacetate (DCFH-DA). Early induction of DNA strand breaks and oxidative DNA base lesions was assessed using the 8-hydroxyguanine DNA-glycosylase 1 (hOGG1)-modified alkaline comet assay, whereas mutagenicity of CQ was determined in the mouse lymphoma TK assay (MLA), according to OECD Guideline No. 490. CQ (0.5-2.5mM) induced concentration- and time-dependent inhibition of cell growth associated with increased ROS/RNS production, amounting to 2342%±1108% of controls after 90min at 2.5mM. Additionally, CQ concentration-dependently caused direct DNA-damage, i.e. formation of DNA strand breaks and 8-hydroxy-2'-deoxyguanosine. Whereas the MLA indicated lack of mutagenicity of CQ after a 4h of treatment, CQ concentration-dependently increased total mutant frequency (MF) after 24h (about 2-fold at 2.5mM). But, based on the global evaluation factor concept, increase in MF did not reach biologically relevance. CQ induced concentration-dependent, cytotoxic and genotoxic effects in L5178Y/TK +/- cells, most likely due to oxidative stress, but without mediating obvious biological relevant mutagenicity. Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  8. Increased heterologous production of the antitumoral polyketide mithramycin A by engineered Streptomyces lividans TK24 strains.

    Science.gov (United States)

    Novakova, Renata; Núñez, Luz Elena; Homerova, Dagmar; Knirschova, Renata; Feckova, Lubomira; Rezuchova, Bronislava; Sevcikova, Beatrica; Menéndez, Nuria; Morís, Francisco; Cortés, Jesús; Kormanec, Jan

    2018-01-01

    Mithramycin A is an antitumor compound used for treatment of several types of cancer including chronic and acute myeloid leukemia, testicular carcinoma, hypercalcemia and Paget's disease. Selective modifications of this molecule by combinatorial biosynthesis and biocatalysis opened the possibility to produce mithramycin analogues with improved properties that are currently under preclinical development. The mithramycin A biosynthetic gene cluster from Streptomyces argillaceus ATCC12956 was cloned by transformation assisted recombination in Saccharomyces cerevisiae and heterologous expression in Streptomyces lividans TK24 was evaluated. Mithramycin A was efficiently produced by S. lividans TK24 under standard fermentation conditions. To improve the yield of heterologously produced mithramycin A, a collection of derivative strains of S. lividans TK24 were constructed by sequential deletion of known potentially interfering secondary metabolite gene clusters using a protocol based on the positive selection of double crossover events with blue pigment indigoidine-producing gene. Mithramycin A production was evaluated in these S. lividans strains and substantially improved mithramycin A production was observed depending on the deleted gene clusters. A collection of S. lividans strains suitable for heterologous expression of actinomycetes secondary metabolites were generated and efficient production of mithramycin A with yields close to 3 g/L, under the tested fermentation conditions was achieved using these optimized collection of strains.

  9. Inhibition of MerTK increases chemosensitivity and decreases oncogenic potential in T-cell acute lymphoblastic leukemia

    International Nuclear Information System (INIS)

    Brandao, L N; Winges, A; Christoph, S; Sather, S; Migdall-Wilson, J; Schlegel, J; McGranahan, A; Gao, D; Liang, X; DeRyckere, D; Graham, D K

    2013-01-01

    Pediatric leukemia survival rates have improved dramatically over the past decades. However, current treatment protocols are still largely ineffective in cases of relapsed leukemia and are associated with a significant rate of chronic health conditions. Thus, there is a continued need for new therapeutic options. Here, we show that mer receptor tyrosine kinase (MerTK) was abnormally expressed in approximately one half of pediatric T-cell leukemia patient samples and T-cell acute lymphoblastic leukemia (T-ALL) cell lines. Stimulation of MerTK by the ligand Gas6 led to activation of the prosurvival proteins Erk 1/2 and Stat5, and MerTK-dependent activation of the STAT pathway in leukemia represents a novel finding. Furthermore, inhibition of MerTK expression increased the sensitivity of T-ALL cells to treatment with chemotherapeutic agents and decreased the oncogenic potential of the Jurkat T-ALL cell line in a methylcellulose colony-forming assay. Lastly, inhibition of MerTK expression significantly increased median survival in a xenograft mouse model of leukemia (30.5 days vs 60 days, P<0.0001). These results suggest that inhibition of MerTK is a promising therapeutic strategy for the treatment of leukemia and may allow for dose reduction of currently used chemotherapeutics resulting in decreased rates of therapy-associated toxicities

  10. UPAYA MENINGKATKAN KREATIVITAS ANAK DENGAN MEMANFAATKAN MEDIA BARANG BEKAS DI TK KOTA BIMA

    Directory of Open Access Journals (Sweden)

    Sri Hardiningsih Hanafi

    2015-12-01

    Full Text Available Penelitian ini bertujuan untuk memperbaiki proses pembelajaran anak TK serta untuk meningkatkan kreativitas anak melalui pemanfaatan media barang bekas di TK N Pembina Kota Bima. Penelitian ini merupakan penelitian tindakan kelas, yang terdiri atas dua siklus. Subjek dari penelitian ini adalah 21 anak. Sedangkan objeknya adalah media daur ulang. Teknik pengambilan data diambil dengan lembar observasi dan catatan lapangan. Sedangkan instrumen yang digunakan adalah lembaran observasi kreativitas anak dan aktivitas guru selama proses pembelajaran. Terjadi peningkatan kreativitas anak secara keseluruhan pada pratindakan dengan rerata 28,81 dengan presentase 19,05%. Rerata pada siklus I adalah 83,85 dengan persentase sebesar 73,73%, sedangkan rerata pada siklus II adalah 135,17 dengan persentase sebesar 87,97%. Sementara itu, aktivitas kegiatan guru mengalami peningkatan dengan rerata sebesar 72,22 pada siklus I dan 94,44 pada siklus II. Dengan demikian media barang bekas dapat meningkatkan kemampuan kreativitas anak dalam proses pembelajaran. Kata Kunci: aspek kreativitas, barang bekas, anak TK kelompok B   IMPROVING CHILDREN’S CREATIVITY THROUGH RECYCLING MEDIA IN TK BIMA CITY Abstract This study aims to improve the learning process of children in kindergartens as well as to enhance the creativity of children through the use of recycling media in TK N Pembina Bima City. This study is a classroom action-research which consisted of two cycles. The subjects of this study were 21 children. While the objects were recycling media. The data collection techniques were observation and field notes while the instruments used were observation sheets of children’s creativity and teacher’s activities during the learning process. There is an overal improvement in children’s creativity after the treatment with a mean of 28.81 and a precentage of 19.05%. The mean of cycle I is 83.85 with a precentage of 73.73% while the mean of cycle II is 135.17 with a

  11. Studies of quaternary deposits in Olkiluoto in 2004; Maaperaetutkimukset tutkimuskaivannoissa TKB ja TK9 Olkiluodossa 2004

    Energy Technology Data Exchange (ETDEWEB)

    Huhta, P. [Geological Survey of Finland, Espoo (Finland)

    2005-04-15

    The study sections (trenches) TK 8 and TK 9 in Olkiluoto were studied according to the contract in autumn 2004. The investigations were carried out in three phases as the excavation progressed. Samples for grain size determinations were taken from vertical profiles, placed 25 m apart along the trench, with the profiles extending from the soil surface down to bedrock or, if the bedrock was covered with water, to the water table. Between 6 and 7 kg of sediment was taken in a bucket with a shovel and placed in plastic bags for transport to the laboratory. The samples were first dried in the laboratory after which they were sieved. In addition, grain size distribution of the < 63 {mu}m fraction was analyzed with the Micromeritics 5000 ET Sedigraph. Sedimentological observations of the sampling profiles were documented in field by drawing (on a field observation form) and the profiles were photographed using a digital camera. In addition, the excavated sections were photographed along their whole length. The length of the TK 8 trench is approximately 730 m and 30 profiles were sampled from it. The height of the profiles varied from 6 m to 0.8 m with an average of 2.6 m. The till in the trench TK 8 is classified as having a normal amount of cobbles and boulders with the size of the larger clasts varying from 6 to 60 cm. The clasts are normally rounded or sub-angular. Both the size and number of cobbles increases with depth, as does their angularity. In places the till is overlain by postglacial clays/silts and littoral sands. With the exception of one sample, all samples from the TK 8 section can be classified as sandy tills based on the d50 grain size. According to the guidelines for mapping of Quatemary deposits one third of the samples is classified as fine-grained tills. This is because of the high percentages of silt and clay size particles that affect the frost heaving properties of the tills as well as their suitability for construction. A till is considered fine

  12. A zinc-dependent protease AMZ-tk from a thermophilic archaeon is a new member of the archaemetzincin protein family

    Directory of Open Access Journals (Sweden)

    Baolei eJia

    2015-12-01

    Full Text Available A putative zinc-dependent protease (TK0512 in Thermococcus kodakarensis KOD1 shares a conserved motif with archaemetzincins, which are metalloproteases found in archaea, bacteria, and eukarya. Phylogenetic and sequence analyses showed that TK0512 and its homologues in Thermococcaceae represent new members in the archaemetzincins family, which we named AMZ-tk. We further confirmed its proteolytic activity biochemically by overexpression of the recombinant AMZ-tk in E. coli and characterization of the purified enzyme. In the presence of zinc, the purified enzyme degraded casein, while adding EDTA strongly inhibited the enzyme activity. AMZ-tk also exhibited self-cleavage activity that required Zn2+. These results demonstrated that AMZ-tk is a zinc-dependent protease within the archaemetzincin family. The enzyme displayed activity at alkaline pHs ranging from 7.0-10.0, with the optimal pH being 8.0. The optimum temperature for the catalytic activity of AMZ-tk was 55ºC. Quantitative reverse transcription-PCR revealed that transcription of AMZ-tk was also up-regulated after exposing the cells to 55 ºC and 65ºC. Mutant analysis suggests that Zn2+ binding histidine and catalytic glutamate plays key roles in proteolysis. AMZ-tk was thermostable on incubation for 4 h at 70°C in the presence of EDTA. AMZ-tk also retained >50% of its original activity in the presence of both laboratory surfactants and commercial laundry detergents. AMZ-tk further showed antibacterial activity against several bacteria. Therefore, AMZ-tk is of considerable interest for many purposes in view of its activity at alkaline pH, detergents, and thermostability.

  13. Small animal PET imaging of HSV1-tk gene expression with {sup 124}IVDU in liver by the hydrodynamic injection

    Energy Technology Data Exchange (ETDEWEB)

    Song, I. H.; Lee, T. S.; Woo, S. G.; Jeong, J. H.; Kang, J. H.; Kim, K. M.; Chun, K. J.; Choi, C. W.; Lim, S. M. [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2007-07-01

    The liver is an important target organ for gene transfer due to its capacity for synthesizing serum protein and its involvement in numerous genetic diseases. High level of foreign gene expression in liver can be achieved by a large-volume and high-speed intravenous injection of naked plasmid DNA (pDNA), so called hydrodynamic injection. This study is aimed to evaluate liver specific-gene expression of herpes simplex virus type 1 thymidine kinase(HSV1-tk) by hydrodynamic injection and image HSV1-tk expression using {sup 124}IVDU-PET. We constructed herpes simplex virus type 1 thymidine kinase (HSV1-tk)-expressing pDNA (pHSV1-tk) modified from pEGFP-N1. Hydrodynamic injection was performed using 40 {mu}g of plasmid (pEGFP/N1 or pHSV1-tk) in 2 ml of 0.85% saline solution for 20{approx}22g mice in 5 seconds intravenously. At 1 d post-hydrodynamic injection, biodistribution study was performed at 2 h post-injection of radiolabeled IVDU, fluorescence image was obtained using optical imager and small animal PET image was acquired with {sup 124}IVDU at 2 h post-injection. After PET imaging, digital whole body autoradiography (DWBA) was performed. Expression of HSV1-tk and EGFP was confirmed by RT-PCR in each liver tissue. In liver of pHSV1-tk and pEGFP/N1 injection groups, {sup 123}IVDU uptake was 5.65%ID/g and 0.98%ID/g, respectively. {sup 123}IVDU uptake in liver of pHSV1-tk injection group showed 5.7-fold higher than that of pEGFP/N1 injection group (p<0.01). On the other hand, the liver of pEGFP/N1 injection group showed fluorescence activity. In small animal PET images, {sup 124}IVDU uptake was selectively localized in liver of pHSV1-tk injection group and also checked in DWBA, but showed minimal uptake in liver of pEGFP/N1 injection mice. Hydrodynamic injection was effective to liver-specific delivery of plasmid DNA. Small animal PET image of {sup 124}IVDU could be used in the evaluation of noninvasive reporter gene imaging in liver.

  14. TkPl_SU: An Open-source Perl Script Builder for Seismic Unix

    Science.gov (United States)

    Lorenzo, J. M.

    2017-12-01

    TkPl_SU (beta) is a graphical user interface (GUI) to select parameters for Seismic Unix (SU) modules. Seismic Unix (Stockwell, 1999) is a widely distributed free software package for processing seismic reflection and signal processing. Perl/Tk is a mature, well-documented and free object-oriented graphical user interface for Perl. In a classroom environment, shell scripting of SU modules engages students and helps focus on the theoretical limitations and strengths of signal processing. However, complex interactive processing stages, e.g., selection of optimal stacking velocities, killing bad data traces, or spectral analysis requires advanced flows beyond the scope of introductory classes. In a research setting, special functionality from other free seismic processing software such as SioSeis (UCSD-NSF) can be incorporated readily via an object-oriented style to programming. An object oriented approach is a first step toward efficient extensible programming of multi-step processes, and a simple GUI simplifies parameter selection and decision making. Currently, in TkPl_SU, Perl 5 packages wrap 19 of the most common SU modules that are used in teaching undergraduate and first-year graduate student classes (e.g., filtering, display, velocity analysis and stacking). Perl packages (classes) can advantageously add new functionality around each module and clarify parameter names for easier usage. For example, through the use of methods, packages can isolate the user from repetitive control structures, as well as replace the names of abbreviated parameters with self-describing names. Moose, an extension of the Perl 5 object system, greatly facilitates an object-oriented style. Perl wrappers are self-documenting via Perl programming document markup language.

  15. Monitoring apoptosis of TK-GFP-expressing ACC-M cells induced by ACV using FRET technique

    Science.gov (United States)

    Xiong, Tao; Zhang, Zhihong; Lin, Juqiang; Yang, Jie; Zeng, Shaoqun; Luo, Qingming

    2006-09-01

    Apoptosis is an evolutionary conserved cellular process that plays an important role during development, but it is also involved in tissue homeostasis and in many diseases. To study the characteristics of suicide gene system of the herpes simplex virus thymidine kinase (HSV-tk) gene in tumor cells and explore the apoptosis phenomena in this system and its effect on the human adenoid cystic carcinoma line ACC-M cell, we detected apoptosis of CD3- (ECFP-CRS-DsRed) and TK-GFP-expressing ACC-M (ACC-M-TK-GFP-CD3) cells induced by acyclovir (ACV) using fluorescence resonance energy transfer (FRET) technique. CD3 is a FRET-based indicator for activity of caspase-3, which is composed of an enhanced cyan fluorescent protein, a caspase-3 sensitive linker, and a red fluorescent protein from Discosoma with efficient maturation property. FRET from ECFP to DsRed could be detected in normal ACC-M-TK-GFP-CD3 cells, and the FRET efficient was remarkably decreased and then disappeared during the cells apoptosis induced by ACV. It was due to the activated caspase-3 cleaved the CD3 fusion protein. In this study, the results suggested that the AVC-induced apoptosis of ACC-M-TK-GFP-CD3 cells was through caspase-3 pathway.

  16. Web, Graphics & PerlTk Programming Best of the Perl Journal

    CERN Document Server

    Orwant, Jon

    2010-01-01

    In its first five years of existence, The Perl Journal (TPJ) became the voice of the Perl community. Every serious Perl programmer subscribed to it, and every notable Perl guru jumped at the opportunity to write for it. TPJ explained critical Perl topics and demonstrated Perl's utility for fields as diverse as astronomy, biology, economics, AI, and games. Back issues were hoarded, or swapped like trading cards. No longer in print format, The Perl Journal remains a proud and timeless achievement of Perl during one of its most exciting periods of development. Web, Graphics & Perl/Tk is the se

  17. Living with Advanced MS

    Science.gov (United States)

    ... Area Donate Donate Search v What Is MS? Definition of MS What Causes MS? Who Gets MS? Multiple Sclerosis FAQs Types of MS ... example, one study found that people with MS who also had a vascular comorbidity (e.g., hypertension, diabetes, high cholesterol) required assistance with walking sooner ...

  18. Wild Type p53 gene sensitizes rat C6 glioma cells to HSV-TK/ACV treatment in vitro and in vivo.

    Science.gov (United States)

    Huang, Qiang; Xia, Zhibo; You, Yongping; Pu, Peiyu

    2010-12-01

    Suicide gene therapy using herpes simplex virus-thymidine kinase (HSV-TK)/ganciclovir (GCV), has been extensively tested for the treatment of glioma. Our previous study showed that exogenous wild type p53 (wt-p53) enhanced the anti-tumor effect of HSV-TK/GCV therapy. However, the use of GCV is hindered by its low penetration to the brain and its toxicity when used at higher dose. In the present study, we used another pro-drug, acyclovir (ACV), and examined the therapeutic efficacy of HSV-TK/ACV combining with wt-p53 in C6 glioma cells. We observed that wt-p53 combined with HSV-TK/ACV resulted in the super-additive anti-tumor effect in vitro. Exogenous wt-p53 significantly enhanced the sensitivity of TK positive C6 cells to ACV in vitro. Our in vivo experiment demonstrated that the effect of wt-p53 and HSV-TK/ACV combination therapy was better than that of HSV-TK/ACV alone. The survival time of tumor-bearing rats treated with wt-p53 in combination with HSV-TK/ACV was also significantly prolonged than those treated with HSV-TK/ACV alone. These results suggest that wt-p53 can enhance the therapeutic efficacy of HSV-TK/ACV both in vitro and in vivo. These findings are considerably valuable with the respect of using less toxic ACV as prodrug. This novel strategy could provide benefit to HSV-TK/prodrug gene therapy.

  19. From the Angle of Reader 50 Year of TKDB (1952-1986 / TK (1987-2001

    Directory of Open Access Journals (Sweden)

    Hasan S. Keseroğlu

    2002-09-01

    Full Text Available This study deals with the conditions and the medium of the issue of the Bulletin of the Turkish Librarians’ Association (TKDB; 1952-1986, which has been na- med as the Journal of the Turkish Librarianship (TK; 1987-2001 later. Besides, the features of it s contents, the changes based on the historical periods and the change it self has been examined closely. This evaluative procedure can be sum- marized as follows: first, the type and the genre of the writings identified as ori- ginal, translation, the latest book reviews, citation/guotation, report, letter, law, argumentative essay, interview,competition or news, considering the features obtained from the close study of each number of t he TKDB / TK j ou mal s within fift.y years. Second, af ter 1980, from the point of scientifi c method and techrıique the numerical data have been obtained fronı the usage of bibliographies, deep rıotes, and the utility of translations into Turkish in the field of Information and documentation, and ali the findings have been discussed.

  20. tkLayout: a Design Tool for Innovative Silicon Tracking Detectors

    CERN Document Server

    Bianchi, Giovanni

    2014-01-01

    A new CMS tracker is scheduled to become operational for the LHC Phase 2 upgrade in the early 2020's. tkLayout is a software package developed to create 3d models for the design of the CMS tracker and to evaluate its fundamental performance figures. The new tracker will have to cope with much higher luminosity conditions, resulting in increased track density, harsher radiation exposure and, especially, much higher data acquisition bandwidth, such that equipping the tracker with triggering capabilities is envisaged. The design of an innovative detector involves deciding on an architecture offering the best trade-off among many figures of merit, such as tracking resolution, power dissipation, bandwidth, cost and so on. Quantitatively evaluating these figures of merit as early as possible in the design phase is of capital importance and it is best done with the aid of software models. tkLayout is a flexible modeling tool: new performance estimates and support for different detector geometries can be quickly ad...

  1. ImTK: an open source multi-center information management toolkit

    Science.gov (United States)

    Alaoui, Adil; Ingeholm, Mary Lou; Padh, Shilpa; Dorobantu, Mihai; Desai, Mihir; Cleary, Kevin; Mun, Seong K.

    2008-03-01

    The Information Management Toolkit (ImTK) Consortium is an open source initiative to develop robust, freely available tools related to the information management needs of basic, clinical, and translational research. An open source framework and agile programming methodology can enable distributed software development while an open architecture will encourage interoperability across different environments. The ISIS Center has conceptualized a prototype data sharing network that simulates a multi-center environment based on a federated data access model. This model includes the development of software tools to enable efficient exchange, sharing, management, and analysis of multimedia medical information such as clinical information, images, and bioinformatics data from multiple data sources. The envisioned ImTK data environment will include an open architecture and data model implementation that complies with existing standards such as Digital Imaging and Communications (DICOM), Health Level 7 (HL7), and the technical framework and workflow defined by the Integrating the Healthcare Enterprise (IHE) Information Technology Infrastructure initiative, mainly the Cross Enterprise Document Sharing (XDS) specifications.

  2. Monitoring of tumor growth and metastasis potential in MDA-MB-435s/tk-luc human breast cancer xenografts

    International Nuclear Information System (INIS)

    Chang, Y.-F.; Lin, Y.-Y.; Wang, H.-E.; Liu, R.-S.; Pang Fei; Hwang, J.-J.

    2007-01-01

    Molecular imaging of reporter gene expression provides a rapid, sensitive and non-invasive monitoring of tumor behaviors. In this study, we reported the establishment of a novel animal model for longitudinal examination of tumor growth kinetics and metastatic spreading in vivo. The highly metastatic human breast carcinoma MDA-MB-435s cell line was engineered to stably express herpes simplex virus type 1 thymidine kinase (HSV-1-tk) and luciferase (luc). Both 131 I-FIAU and D-luciferin were used as reporter probes. For orthotopic tumor formation, MDA-MB-435s/tk-luc cells were implanted into the first nipple of 6-week-old female NOD/SCID mice. For metastatic study, cells were injected via the lateral tail vein. Mice-bearing MDA-MB-435s/tk-luc tumors were scanned for tumor growth and metastatsis using Xenogen IVIS50 system. Gamma scintigraphy and whole-body autoradiography were also applied to confirm the tumor localization. The results of bioluminescence imaging as well as histopathological finding showed that tumors could be detected in femur, spine, ovary, lungs, kidney, adrenal gland, lymph nodes and muscle at 16 weeks post i.v. injection, and correlated photons could be quantified. This MDA-MB-435s/tk-luc human breast carcinoma-bearing mouse model combined with multimodalities of molecular imaging may facilitate studies on the molecular mechanisms of cancer invasion and metastasis

  3. Sequence, Structure, and Binding Analysis of Cyclodextrinase (TK1770 from T. kodakarensis (KOD1 Using an In Silico Approach

    Directory of Open Access Journals (Sweden)

    Ramzan Ali

    2015-01-01

    Full Text Available Thermostable cyclodextrinase (Tk1770 CDase from hyperthermophilic archaeon Thermococcus kodakarensis (KOD1 hydrolyzes cyclodextrins into linear dextrins. The sequence of Tk1770 CDase retrieved from UniProt was aligned with sequences of sixteen CD hydrolyzing enzymes and a phylogenetic tree was constructed using Bayesian inference. The homology model of Tk1770 CDase was constructed and optimized with Modeller v9.14 program. The model was validated with ProSA server and PROCHECK analysis. Four conserved regions and the catalytic triad consisting of Asp411, Glu437, and Asp502 of GH13 family were identified in catalytic site. Also an additional fifth conserved region downstream to the fourth region was also identified. The structure of Tk1770 CDase consists of an additional N′-domain and a helix-loop-helix motif that is conserved in all archaeal CD hydrolyzing enzymes. The N′-domain contains an extended loop region that forms a part of catalytic domain and plays an important role in stability and substrate binding. The docking of substrate into catalytic site revealed the interactions with different conserved residues involved in substrate binding and formation of enzyme-substrate complex.

  4. A predictive toxicogenomics signature to classify genotoxic versus non-genotoxic chemicals in human TK6 cells

    Directory of Open Access Journals (Sweden)

    Andrew Williams

    2015-12-01

    Full Text Available Genotoxicity testing is a critical component of chemical assessment. The use of integrated approaches in genetic toxicology, including the incorporation of gene expression data to determine the DNA damage response pathways involved in response, is becoming more common. In companion papers previously published in Environmental and Molecular Mutagenesis, Li et al. (2015 [6] developed a dose optimization protocol that was based on evaluating expression changes in several well-characterized stress-response genes using quantitative real-time PCR in human lymphoblastoid TK6 cells in culture. This optimization approach was applied to the analysis of TK6 cells exposed to one of 14 genotoxic or 14 non-genotoxic agents, with sampling 4 h post-exposure. Microarray-based transcriptomic analyses were then used to develop a classifier for genotoxicity using the nearest shrunken centroids method. A panel of 65 genes was identified that could accurately classify toxicants as genotoxic or non-genotoxic. In Buick et al. (2015 [1], the utility of the biomarker for chemicals that require metabolic activation was evaluated. In this study, TK6 cells were exposed to increasing doses of four chemicals (two genotoxic that require metabolic activation and two non-genotoxic chemicals in the presence of rat liver S9 to demonstrate that S9 does not impair the ability to classify genotoxicity using this genomic biomarker in TK6cells.

  5. Implementasi Pendidikan Entrepreneurship pada Anak Usia Dini di TK Khalifah Sukonandi Yogyakarta

    Directory of Open Access Journals (Sweden)

    Lailatu Rohmah

    2017-12-01

    Full Text Available Entrepreneurial spirit and entrepreneurship education have grown everywhere. But ideally this spirit is implanted as early as possible. So far there has been no serious kindergarten to support this entrepreneurial spirit. TK Khalifah who has tauhid education and entrepreneurship. This research focuses on: (1 how the implementation of entrepreneurship education in Khalifah kindergarten, (2 entrepreneurship education activities in Khalifah Kindergarten, and (3 factors that support and obstruct early childhood entrepreneurship education in Khalifah Kindergarten. This research is qualitative research, and data collected by interview, documentation, and observation. Data analysis with reduction, display, and verification / conclusion. The results showed that the implementation of entrepreneurship education in early childhood in Khalifah kindergarten is integrating entrepreneurial values in learning.

  6. PinK - a TCL/TK based database interface to ADAMO and DAD

    International Nuclear Information System (INIS)

    Ackerstaff, K.; Funk, M.A.; Dueren, M.; Ferstl, M.; Wander, W.; Welch, P.

    1996-01-01

    PinK is a TCL/TK extension developed initially for the HERMES experiment at HERA/DESY. Like other HEP experiments, HERMES needs a highly structured database to cope with the complexity and amount of data being handled. HERMES uses the ADAMO Entity Relationship database concept together with the DAD package which extends the use of ADAMO towards a distributed database with flexible, fast I/O and time dependent data handling. With a large and complex time dependent database and many programs accessing it, there is a strong demand for a flexible common user interface. PINK incorporates the functionality of ADAMO and DAD providing an Object Oriented interface to the database. It can be used as an interactive command line, as well as graphical user interface to access and manipulate data on the databases serves and files. (author)

  7. Mineralogical study on quaternary soils of the Olkiluoto investigation trench OL-TK19

    Energy Technology Data Exchange (ETDEWEB)

    Lukkari, S.; Marmo, J. [Geological Survey of Finland, Espoo (Finland)

    2014-07-15

    Six till samples representing three vertical profiles in a research trench OL TK19 from Olkiluoto were studied for their mineralogy. The client, Posiva Oy, was interested especially in carbonates and sulphides. The study is based on size and density classified subsamples. Mineral distributions were determined by a Scanning Electron Microscope (SEM) equipped with an energy dispersive X-ray spectrometer (EDS), using the INCA FEATURE phase detection and classification software. Lithologies within the cobble fraction are also measured. The study revealed unhomogeneity in the sample material. Sample TK 19/2 POH is rich in calcareous sedimentary rocks and older till compared to the other samples. Three main carbonate-bearing sedimentary rock types are: limestone, carbonate cemented quartz feldspar rock, and quartz graywacke, containing clasts of the carbonate cemented quartz feldspar rock. The contents of heavy minerals (0.1-1.2%) and semi heavy minerals (1.3-3.3%) are low in every sample. Carbonate concentrates of the semi heavy mineral separates are low (0.04-0.57 %). Sulfide is predominately pyrite (1.3-8.6%). Pyrite grains have commonly goethite crusts, which are ubiquitous in the surface samples. Both sulphide and carbonate occurs as liberated grains and as fine grained matrix, enveloping other mineral grains. Phosphorites and phosphorous rock fragments are also common in the till samples. The mode of occurrence of phosphate resembles that of carbonate and sulphides that makes the quantification of these buffer minerals impossible. Sedimentary rocks are interpreted to have been derived from the sedimentary rock pile preserved under the Gulf of Bothnia. (orig.)

  8. Effect of chemical mutagens and carcinogens on gene expression profiles in human TK6 cells.

    Directory of Open Access Journals (Sweden)

    Lode Godderis

    Full Text Available Characterization of toxicogenomic signatures of carcinogen exposure holds significant promise for mechanistic and predictive toxicology. In vitro transcriptomic studies allow the comparison of the response to chemicals with diverse mode of actions under controlled experimental conditions. We conducted an in vitro study in TK6 cells to characterize gene expression signatures of exposure to 15 genotoxic carcinogens frequently used in European industries. We also examined the dose-responsive changes in gene expression, and perturbation of biochemical pathways in response to these carcinogens. TK6 cells were exposed at 3 dose levels for 24 h with and without S9 human metabolic mix. Since S9 had an impact on gene expression (885 genes, we analyzed the gene expression data from cells cultures incubated with S9 and without S9 independently. The ribosome pathway was affected by all chemical-dose combinations. However in general, no similar gene expression was observed among carcinogens. Further, pathways, i.e. cell cycle, DNA repair mechanisms, RNA degradation, that were common within sets of chemical-dose combination were suggested by clustergram. Linear trends in dose-response of gene expression were observed for Trichloroethylene, Benz[a]anthracene, Epichlorohydrin, Benzene, and Hydroquinone. The significantly altered genes were involved in the regulation of (anti- apoptosis, maintenance of cell survival, tumor necrosis factor-related pathways and immune response, in agreement with several other studies. Similarly in S9+ cultures, Benz[a]pyrene, Styrene and Trichloroethylene each modified over 1000 genes at high concentrations. Our findings expand our understanding of the transcriptomic response to genotoxic carcinogens, revealing the alteration of diverse sets of genes and pathways involved in cellular homeostasis and cell cycle control.

  9. Responses of the L51781Y tk/sup +//tk/sup -/ mouse lymphoma cell forward mutation assay: III. 72 coded chemicals

    Energy Technology Data Exchange (ETDEWEB)

    McGregor, D.B.; Brown, A.; Cattanach, P.; Edwards, I.; McBride, D.; Riach, C.; Caspary, W.J.

    1988-01-01

    Seventy-two chemicals were tested for their mutagenic potential in the L51781Y tk/sup +///sup -/ mouse lymphoma cell forward mutation assay, using procedures based upon those described previously. Cultures were exposed to the chemicals for 4 hr, then cultured for 2 days before planting in soft agar with or without trifluorothymidine (TFT), 3 ..mu..g/ml. The chemicals were tested at least twice. Significant responses were obtained with allyl isothiocyanate, p-benzoquinone dioxime, benzyl acetate, 2-biphenylamine HCl, bis(2-chloro-1-methylethyl)ether, cadmium chloride, chlordane, chlorobenzene, chlorobenzilate, 2-chloroethanol, chlorothalonil, cytarabine x HCl, p,p'-DDE, diazinon, 2,6-dichloro-p-phenylenediamine, N,N-diethylthiourea, diglycidylresorcinol ether, 2,4-dimethoxy aniline x HCl, disperse yellow 3, endosulfan, 1,2-epoxyhexadecane, ethyl acrylate, ethyl benzene, ethylene thiourea, F D and C yellow Number 6, furan, heptachlor, isophorone, mercuric chloride, 4,4'-methylenedianiline x 2 HCl, methyl viologen, nickel sulfate x 6H/sub 2/O, 4,4'-oxydianiline, pentachloroethane, piperonyl butoxide, propyl gallate, quinoline, rotenone, 2,4,5,6-tetrachloro-4-nitro-anisole, 1,1,1,2-tetrachloroethane, trichlorfon, 2,4,6-trichlorophenol, 2,4,5-trimethoxybenzaldehyde, 1,1,3-trimethyl-2-thiourea, 1-vinyl-3-cyclopetene dioxide, vinyl toluene, and ziram. The assay was incapable of providing a clear indication of whether some chemicals were mutagens; these benzyl alcohol, 1,4-dichlorobenzene, phenol, succinic acid-2,2-dimethyl hydrazide, and toluene.

  10. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... MS Symptoms Diagnosing MS Possible MS Clinically Isolated Syndrome (CIS) Newly Diagnosed Diagnosing Tools Other Conditions to ... Sclerosis FAQs d Types of MS Clinically Isolated Syndrome (CIS) Relapsing-remitting MS (RRMS) Primary progressive MS ( ...

  11. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Area Donate Donate Search v What Is MS? Definition of MS What Causes MS? Who Gets MS? ... Support Personal Stories d What Is MS? d Definition of MS Myelin Immune-Mediated Disease T Cells ...

  12. In vivo and in vitro experimental study on cervix cancer with combination of HSV-TK/GCV suicide gene therapy system and 60Co radiotherapy

    International Nuclear Information System (INIS)

    Chen Daozhen; Xue Wenqun; Zhan Huiying; Zhu Yunxia; Yang Youyi; Liu Lu; Tang Qiusha

    2006-01-01

    Objective: To evaluate the killing effect of HSV-TK/GCV suicide gene therapy system combined with 60 Co radiotherapy on human cervical cancer HeLa cell line in vivo and in vitro, and to explore radiosensitization by the HSV-TK/GCV system. Methods: The HSV-TK/GCV suicide gene therapy system and 60 Co radiotherapy were used separately or in combination for human cervical cancer HeLa cell line in vivo and in vitro to compare their effects. Colony formation test and the rate of radiosensitization effect(E/O) were employed to observed the radiosensitization by the HSV-TK/GCV system. Results: The HSV-TK/GCV suicide gene therapy system showed strong therapeutic effects on HeLa cells both in vitro and in vivo (the inhibition rates were 45.8% and 39.5%, respectively). Moreover, the combined application of gene therapy and radiotherapy exhibited stronger therapeutic effects in vitro and in vivo (the inhibition rate was 87.5% in vitro, and was 87.9% in vivo) (P 1.4), indicating the HSV-TK/GCV system could exert a sensitizing effect on 60 Co radiotherapy of the transplanted human cervical cancer cells in nude mice. Conclusion: The HSV-TK/GCV system has radiosensitizationaction. Gene therapy combined with radiotherapy may be a good supplementary method for synthetic treatment of cervical cancer. (authors)

  13. Gene expression deregulation in postnatal skeletal muscle of TK2 deficient mice reveals a lower pool of proliferating myogenic progenitor cells.

    Directory of Open Access Journals (Sweden)

    João A Paredes

    Full Text Available Loss of thymidine kinase 2 (TK2 causes a heterogeneous myopathic form of mitochondrial DNA (mtDNA depletion syndrome (MDS in humans that predominantly affects skeletal muscle tissue. In mice, TK2 deficiency also affects several tissues in addition to skeletal muscle, including brain, heart, adipose tissue, kidneys and causes death about 3 weeks after birth. We analysed skeletal muscle and heart muscle tissues of Tk2 knockout mice at postnatal development phase and observed that TK2 deficient pups grew slower and their skeletal muscles appeared significantly underdeveloped, whereas heart was close to normal in size. Both tissues showed mtDNA depletion and mitochondria with altered ultrastructure, as revealed by transmission electron microscopy. Gene expression microarray analysis showed a strong down-regulation of genes involved in cell cycle and cell proliferation in both tissues, suggesting a lower pool of undifferentiated proliferating cells. Analysis of isolated primary myoblasts from Tk2 knockout mice showed slow proliferation, less ability to differentiate and signs of premature senescence, even in absence of mtDNA depletion. Our data demonstrate that TK2 deficiency disturbs myogenic progenitor cells function in postnatal skeletal muscle and we propose this as one of the causes of underdeveloped phenotype and myopathic characteristic of the TK2 deficient mice, in addition to the progressive mtDNA depletion, mitochondrial damage and respiratory chain deficiency in post-mitotic differentiated tissue.

  14. STAT3/NF-κB-Regulated Lentiviral TK/GCV Suicide Gene Therapy for Cisplatin-Resistant Triple-Negative Breast Cancer.

    Science.gov (United States)

    Kuo, Wei-Ying; Hwu, Luen; Wu, Chun-Yi; Lee, Jhih-Shian; Chang, Chi-Wei; Liu, Ren-Shyan

    2017-01-01

    Triple-negative breast cancer (TNBC) represents approximately 20% of all breast cancers and appears resistance to conventional cytotoxic chemotherapy, demonstrating a particularly poor prognosis and a significantly worse clinical outcome than other types of cancer. Suicide gene therapy has been used for the in vivo treatment of various solid tumors in recent clinical trials. In tumor microenvironment, STAT3/NF-κB pathways are constitutively activated in stromal cells as well as in cancer stem cells (CSCs). In this study, we have cloned a novel STAT3/NF-κB-based reporter system to drive the expression of herpes simplex virus thymidine kinase (HSV-TK) against breast cancer. Lentiviral vector expressing HSV-TK under the regulation of STAT3/NF-κB fused response element was developed. In this setting, we exploited the constitutive STAT3/NF-κB activation in tumors to achieve higher transgene expression than that driven by a constitutively active CMV promotor in vivo . An orthotropic MDA-MB-231 triple negative breast cancer mouse model was used for evaluating the feasibility of STAT3-NF-κB-TK/GCV suicide gene therapy system. The basal promoter activity of Lenti-CMV-TK and Lenti-STAT3-NF-κB-TK in MDA-MB-231 cells was compared by 3 H-FEAU uptake assay. The Lenti-CMV-TK showed ~5 fold higher 3 H-FEAU uptake then Lenti -STAT3-NF-κB-TK. In clonogenic assay, cells expressing Lenti-CMV-TK were 2-fold more sensitive to GCV than Lenti-STAT3-NF-κB-TK transduced cells. In vitro effect of STAT3-NF-κB-induced transgene expression was determined by 10ng/mL TNF-α induction and confirmed by western blot analysis and DsRedm fluorescent microscopy. In vivo evaluation of therapeutic effect by bioluminescence and [ 18 F]FHBG microPET imaging indicated that Lenti-STAT3-NF-κB-TK showed more tumor growth retardation than Lenti-CMV-TK when GCV (20 mg/kg) was administered. The invasiveness and expression of cancer stem cell markers were both decreased after STAT3/NF-κB-regulated HSV-TK

  15. Comparison of [18F]FHBG and [14C]FIAU for imaging of HSV1-tk reporter gene expression: adenoviral infection vs stable transfection

    International Nuclear Information System (INIS)

    Min, Jung-Jun; Iyer, Meera; Gambhir, Sanjiv S.

    2003-01-01

    Earlier studies involving comparison of different reporter probes have shown conflicting results between pyrimidine nucleosides [e.g., 2'-fluoro-2'-deoxy-1-β-d-arabinofuranosyl-5-iodouracil (FIAU)] and acycloguanosine derivatives [e.g., penciclovir (PCV), 9-(4-fluoro-3-hydroxymethylbutyl)guanine (FHBG)]. We hypothesized that this reported discrepancy may be related to how the reporter gene is delivered to the cells - stably transfected vs adenoviral infection. We directly compared the uptake characteristics of [ 18 F]FHBG, [ 3 H]PCV, and [ 14 C]FIAU in cell culture and in vivo using an adenoviral mediated gene transfer model and stably transfected cells. We further compared the uptake of three reporter probes using both HSV1-tk and a mutant HSV1-sr39tk expressing cells to assess the optimal reporter probe/reporter gene combination. [ 14 C]FIAU accumulation was greater than that of [ 3 H]PCV and [ 18 F]FHBG in control cells and in HSV1-tk stably transfected cells (P 8 pfu), [ 18 F]FHBG and [ 3 H]PCV accumulation was significantly greater than that of [ 14 C]FIAU (P 18 F]FHBG and [ 3 H]PCV accumulated to a significantly greater extent than [ 14 C]FIAU in C6-stb-sr39tk+ and AdCMV-HSV1-sr39tk infected C6 cells (P 14 C]FIAU led to significantly higher %ID/g in C6-stb-tk+ xenografts than [ 18 F]FHBG (P 14 C]FIAU, [ 18 F]FHBG led to as high %ID/g in HSV1-tk expressing hepatocytes and to significantly greater %ID/g in C6-stb-sr39tk+ xenografts and HSV1-sr39tk expressing hepatocytes. Dynamic sequential images showed that [ 18 F]FHBG was well retained in HSV1-sr39tk expressing cells (C6-stb-sr39tk+) for at least 4 h after injection, while it was rapidly cleared from HSV1-tk expressing cells (MH3924A-stb-tk+). [ 14 C]FIAU accumulated in HSV1-tk stably expressing cells to a greater extent than either [ 3 H]PCV or [ 18 F]FHBG. However, the accumulation of [ 3 H]PCV and [ 18 F]FHBG in adenoviral infected C6 cells or hepatocytes was equivalent to or greater than that of [ 14 C

  16. Pengembangan Peta Tiga Dimensi Interaktif untuk Dharma Wanita, TK dan Wisma Yasmine Institut Teknologi Sepuluh Nopember Menggunakan Unreal Engine

    Directory of Open Access Journals (Sweden)

    Mochamad Rahman Irwandi

    2013-12-01

    Full Text Available Peningkatan teknologi yang berkembang pesat saat ini berdampak pada penyajian informasi mengenai bangunan, yang dahulu menggunakan dua dimensi (2D, tetapi sekarang menggunakan tiga dimensi (3D. Tampilan gambar 3D ini membuat bangunan terlihat lebih detail dan menarik, tak terkecuali pada perguruan tinggi Institut Teknologi Sepuluh Nopember (ITS yang menjadi objek visualisasi 3D sebagai sarana promosi pada konsumen.Pada tugas akhir ini penulis mengembangkan aplikasi peta tiga dimensi dengan menggunakan Unreal Development Kit (UDK Engine. Aplikasi peta 3D ini menyajikan tampilan Gedung Dharma Wanita, TK dan Wisma Yasmine ITS secara interaktif dan akurat, sehingga pengguna mendapatkan pengalaman yang berbeda dengan mengetahui gedung Dharma Wanita, TK dan Wisma Yasmine ITS secara virtual tanpa harus pergi ke tempat tersebut

  17. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... MS? d Definition of MS Myelin Immune-Mediated Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters d Who Gets MS? Pediatric MS African Americans Hispanics & Latino/ ...

  18. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... v What Is MS? Definition of MS What Causes MS? Who Gets MS? Multiple Sclerosis FAQs Types ... Myelin Immune-Mediated Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters d Who Gets ...

  19. Both p53-PUMA/NOXA-Bax-mitochondrion and p53-p21cip1 pathways are involved in the CDglyTK-mediated tumor cell suppression

    International Nuclear Information System (INIS)

    Yu, Zhendong; Wang, Hao; Zhang, Libin; Tang, Aifa; Zhai, Qinna; Wen, Jianxiang; Yao, Li; Li, Pengfei

    2009-01-01

    CDglyTK fusion suicide gene has been well characterized to effectively kill tumor cells. However, the exact mechanism and downstream target genes are not fully understood. In our study, we found that CDglyTK/prodrug treatment works more efficiently in p53 wild-type (HONE1) cells than in p53 mutant (CNE1) cells. We then used adenovirus-mediated gene delivery system to either knockdown or overexpress p53 and its target genes in these cells. Consistent results showed that both p53-PUMA/NOXA/Bcl2-Bax and p53-p21 pathways contribute to the CDglyTK induced tumor cell suppression. Our work for the first time addressed the role of p53 related genes in the CDglyTK/prodrug system.

  20. Development of a sufficient and effective procedure for transformation of an oleaginous yeast, Rhodosporidium toruloides DMKU3-TK16.

    Science.gov (United States)

    Tsai, Yung-Yu; Ohashi, Takao; Kanazawa, Takenori; Polburee, Pirapan; Misaki, Ryo; Limtong, Savitree; Fujiyama, Kazuhito

    2017-05-01

    Rhodosporidium toruloides DMKU3-TK16 (TK16), a basidiomycetous yeast isolated in Thailand, can produce a large amount of oil corresponding to approximately 70 % of its dry cell weight. However, lack of a sufficient and efficient transformation method makes further genetic manipulation of this organism difficult. We here developed a new transformation system for R. toruloides using a lithium acetate method with the Sh ble gene as a selective marker under the control of the R. toruloides ATCC 10657 GPD1 promoter. A linear DNA fragment containing the Sh ble gene expression cassette was integrated into the genome, and its integration was confirmed by colony PCR and Southern blot. Then, we further optimized the parameters affecting the transformation efficiency, such as the amount of linear DNA, the growth phase, the incubation time in the transformation mixture, the heat shock treatment temperature, the addition of DMSO and carrier DNA, and the recovery incubation time. With the developed method, the transformation efficiency of approximately 25 transformants/μg DNA was achieved. Compared with the initial trial, transformation efficiency was enhanced 417-fold. We further demonstrated the heterologous production of EGFP in TK16 by microscopic observation and immunoblot analysis, and use the technique to disrupt the endogenous URA3 gene. The newly developed method is thus simple and time saving, making it useful for efficient introduction of an exogenous gene into R. toruloides strains. Accordingly, this new practical approach should facilitate the molecular manipulation, such as target gene introduction and deletion, of TK16 and other R. toruloides strains as a major source of biodiesel.

  1. Criticality safety analysis of TK-13 cask in Bushehr nuclear power plant

    Energy Technology Data Exchange (ETDEWEB)

    Mohammadi, Ashgar; Omidvari, Nima [Iran Radioactive Waste Management Company, Tehran (Iran, Islamic Republic of); Hassanzadeh, Mostafa [Nuclear Science and Technology Research Institute, Tehran (Iran, Islamic Republic of)

    2017-12-15

    Spent fuel production is one of the main problems of nuclear power plants that should be managed properly considering the strategy of each country. Today, in most of nuclear power owner countries, the interim storage has been selected as the temporary solution of spent fuel management because of absence of deep geological repositories and no tendency for reprocessing. On the other side, considering the merits of storage in dual purpose casks based on dry storage, this method was chosen for interim storage. By taking into account that the only operating reactor of Iran is of Water-Water Energetic Reactor (WWER)-1000 type, proposed TK-13 cask by Russia which is the manufacturer of these types of reactors has been considered. In this study, the calculation of basket holding spent fuel assembly criticality of this cask has been analyzed for two modes of fresh and spent fuel by ORIGEN2.1 and MCNPX2.6 nuclear codes. The criterion of the nuclear criticality safety for effective multiplication factor (k{sub eff}) should be 0.95 and 0.98 for many ordinary and accident conditions, respectively. Therefore, the results show that a cylindrical basket with 66 cm diameter and 28 cm pitch with internal holding basket made of borated steel with 0.1% borate and steel free from borate would meet the criticality of cask, respectively.

  2. Enzymatic activity of a novel halotolerant lipase from Haloarcula hispanica 2TK2

    Directory of Open Access Journals (Sweden)

    Ozgen Melis

    2016-06-01

    Full Text Available A strain of Haloarcula hispanica isolated from Tuzkoy salt mine, Turkey exhibited extracellular lipolytic activity. Important parameters such as carbon sources and salt concentration for lipase production were investigated. Optimal conditions for the enzyme production from Haloarcula hispanica 2TK2 were determined. It was observed that the lipolytic activity of Haloarcula hispanica was stimulated by some of the carbon sources. The high lipase acitivity values were obtained in the presence of 2% (v/v walnut oil (6.16 U/ml, 1% (v/v fish oil (5.07 U/ml, 1% (v/v olive oil (4.52 U/ml and 1% (w/v stearic acid (4.88 U/ml at 4M NaCl concentration. Lipase was partially purified by ammonium sulfate precipitation and ultrafiltration. Optimal temperature and pH values were determined as 45°C and 8.0, respectively. Lipase activity decreased with the increasing salt concentration, but 85% activity of the enzyme was maintained at 5M NaCl concentration. The enzyme preserved 41% of its relative activity at 90°C. The partially purified lipase maintained its activity in the presence of surfactants such as Triton X-100 and SDS. Therefore, the lipase which is an extremozyme may have potential applications especially in detergent industry.

  3. Complete genome sequence of Hydrogenobacter thermophilus type strain (TK-6T)

    Energy Technology Data Exchange (ETDEWEB)

    Zeytun, Ahmet [Los Alamos National Laboratory (LANL); Sikorski, Johannes [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Nolan, Matt [Joint Genome Institute, Walnut Creek, California; Lapidus, Alla L. [Joint Genome Institute, Walnut Creek, California; Lucas, Susan [Joint Genome Institute, Walnut Creek, California; Han, James [Joint Genome Institute; Tice, Hope [Joint Genome Institute, Walnut Creek, California; Cheng, Jan-Fang [Joint Genome Institute, Walnut Creek, California; Tapia, Roxanne [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [Joint Genome Institute, Walnut Creek, California; Liolios, Konstantinos [Joint Genome Institute, Walnut Creek, California; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Mikhailova, Natalia [U.S. Department of Energy, Joint Genome Institute; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Chen, Amy [Joint Genome Institute, Walnut Creek, California; Palaniappan, Krishna [Joint Genome Institute, Walnut Creek, California; Ngatchou, Olivier Duplex [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Chang, Yun-Juan [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Han, Cliff [Los Alamos National Laboratory (LANL); Detter, J. Chris [Joint Genome Institute, Walnut Creek, California; Ubler, Susanne [Universitat Regensburg, Regensburg, Germany; Rohde, Manfred [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Tindall, Brian [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Wirth, Reinhard [Universitat Regensburg, Regensburg, Germany; Woyke, Tanja [Joint Genome Institute, Walnut Creek, California; Bristow, James [Joint Genome Institute, Walnut Creek, California; Eisen, Jonathan [Joint Genome Institute, Walnut Creek, California; Markowitz, Victor [Joint Genome Institute, Walnut Creek, California; Hugenholtz, Philip [U.S. Department of Energy, Joint Genome Institute; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Kyrpides, Nikos C [Joint Genome Institute, Walnut Creek, California

    2011-01-01

    Hydrogenobacter thermophilus Kawasumi et al. 1984 is the type species of the genus Hydrogenobacter. H. thermophilus was the first obligate autotrophic organism reported among aerobic hydrogen-oxidizing bacteria. Strain TK-6T is of interest because of the unusually efficient hydrogen-oxidizing ability of this strain, which results in a faster generation time compared to other autotrophs. It is also able to grow anaerobically using nitrate as an electron acceptor when molecular hydrogen is used as the energy source, and able to aerobically fix CO2 via the reductive tricarboxylic acid cycle. This is the fifth completed genome sequence in the family Aquificaceae, and the second genome sequence determined from a strain derived from the original isolate. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 1,742,932 bp long genome with its 1,899 protein-coding and 49 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  4. Synthesis of substrates for gene therapy monitoring of HSV1-TK system

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Tae Hyun; Ahn, Soon Hyuk; Choi, Chang Woon; Lim, Sang Moo; Awh, Ok Doo [College of Medicine, Yonsei Univ., Wonju (Korea, Republic of)

    2002-04-01

    In gene therapy, tumor cells expressing the herpes simplex virus thymidine kinase are sensitive to prodrugs. Potential prodrugs IVDU and IVFRU were synthesized and radiolabeled with radioiodine for noninvasive imaging of herpes simplex virus type 1 gene expression. 5-(2-trimethysilyl) vinyl-2'-deoxyuridine and 5-t(2-trimethylsilyl)vinyl-2'-fluoro-2'-deoxyuridine, precursors of 5-(2-iodo)viny 1-2'-deoxy uridine(IVDU) and 5-(2-iodo)-2'-vinyl-2'-deoxy-2'-fluorotibofuranosyl uracil(IVFRU), were synthesized from reaction of trans-1-trimethylsillyl-2-tri-n-butylstannylethylene with 5-iodo-2'-deoxyuridine and 5-iodo-2'-fluoro-2'-deoxyuridine, respectively, on the condition of Pd catalyst. These precursors were separated from reaction mixture by silica gel column chromatography method. Each precursor was radioiodinated with radioiodine by mixing with ICI oxidizing agent. These radioiodinated compounds were purified with HPLC. Radiohalogen exchange has been shown to be effective for the synthesis of products with lower specific activity. Similarly, carrier-added and high specific activity products have been isolated in respectable radiochemical yields using ICI method. Synthetic yield of precursors, IVDU and IVFRU were 43% and 18%, respectively. Radiochemical purity of both compunds was over 98%. We synthesized precursors of IVDU and IVFRU for monitoring of HSV1-tk gene expression. Radiotracers were radioiodinated with high radiolabeling yield by ICI method.

  5. From the Angle of Reader 50 Year of TKDB (1952-1986 / TK (1987-2001 Okur Gözüyle TKDB (1952-1986 / TK (1987-2001 Dergisinin 50 Yılı

    Directory of Open Access Journals (Sweden)

    Hasan S. Keseroğlu

    2002-09-01

    Full Text Available This study deals with the conditions and the medium of the issue of the Bulletin of the Turkish Librarians' Association (TKDB; 1952-1986, which has been named as the Journal of the Turkish Librarianship (TK; 1987-2001 later. Besides, the features of its contents, the changes based on the historical periods and the change itself has been examined closely. This evaluative procedure can be summarized as follows: first, the type and the genre of the writings identified as original, translation, the latest book reviews, citation/quotation, report, letter, law, argumentative essay, interview, competition or news, considering the features obtained from the close study of each number of the TKDB / TK journals within fifty years. Second, after 1980, from the point of scientific method and technique the numerical data have been obtained from the usage of bibliographies, deep notes, and the utility of translations into Turkish in the field of information and documentation, and all the findings have been discussed. Bu çalışmada, Türk Kütüphaneciler Derneği Bülteni (TKDB; 1952-1986 değişen adıyla Türk Kütüphaneciliği (TK; 1987-2001 dergisinin çıkış koşulları ve ortamı anlamaya çalışılmakta; ayrıca TKDB /TK'nin içerik özellikleri, dönemlere bağlı olarak değişimi, bu değişimin irdelenmesi ele alınmaktadır.Bu değerlendirme işlemi, öncesinde TKDB / TK dergilerinin elli yıllık sayılarındaki yazılar tek tek gözden geçirilerek özellikleri tür (telif, çeviri, yayın tanıtım, alıntı/aktarma, rapor, mektup, yasa, tartışma metni, röportaj, yarışma ve haber olarak; 1980 sonrası da bilimsel yöntem ve teknik açısından kaynakça, dipnot kullanma ve bilgi ve belge yönetimi konusunda Türkçeye yapılan çevirilerden yararlanma yönünde sayısal veriler belirlenmiş ve bu veriler tartışılmıştır.

  6. An experimental study on cervix cancer with combination of HSV-TK/GCV suicide gene therapy system and 60Co radiotherapy

    International Nuclear Information System (INIS)

    Chen, Daozhen; Tang, Qiusha

    2010-01-01

    To evaluate the killing effect of HSV-TK/GCV suicide gene therapy system combined with 60 Co radiotherapy on human cervical cancer Hela cell line in vitro and in vivo, and to explore the radiosensitization by HSV-TK/GCV system. HSV-TK/GCV suicide gene therapy system and 60 Co radiotherapy were used separately or in combination on human cervical cancer Hela cell line in vitro and in vivo to compare their effects. Colony formation test and the rate of radiosensitization effect (E/O) were employed to observed the radiosensitization by HSV-TK/GCV system. HSV-TK/GCV suicide gene therapy system had strong therapeutic effects on Hela cells in vitro and in vivo (the inhibition rates were 45.8% and 39.5%, respectively), moreover, the combined administration of gene therapy and radiotherapy had stronger therapeutic effects in vitro and in vivo (the inhibition rate was 87.5% in vitro, and the inhibition rate was 87.9% in vivo) (P < 0.01). The inhibition rate by radiotherapy alone was 42.4% in vitro and 35.8% in vivo. The sensitivity of combined therapy to radiotherapy increased more than that of therapy alone, the ability of colony formation decreased (P < 0.01). The rate of radiosensitivity effect (E/O) was 3.17(> 1.4), indicating HSV-TK/GCV system could exert a sensitizing effect on 60 Co radiotherapy of the transplanted human cervical cancer cell in nude mice. HSV-TK/GCV system had radiosensitization. Gene therapy combined with radiotherapy may be a good supplementary method for cervix cancer synthetic treatment

  7. Non-invasive imaging of cardiac transgene expression with PET: comparison of the human sodium/iodide symporter gene and HSV1-tk as the reporter gene

    Energy Technology Data Exchange (ETDEWEB)

    Miyagawa, Masao; Haubner, Roland; Souvatzoglou, Michael; Schwaiger, Markus; Bengel, Frank M. [Technische Universitaet Muenchen, Klinikum rechts der Isar, Nuklearmedizinische Klinik und Poliklinik, Muenchen (Germany); Anton, Martina; Gansbacher, Bernd [Technische Universitaet Muenchen, Institut fuer Experimentelle Onkologie und Therapieforschung, Munich (Germany); Wagner, Bettina [Technische Universitaet Muenchen, Klinikum rechts der Isar, Nuklearmedizinische Klinik und Poliklinik, Muenchen (Germany); Technische Universitaet Muenchen, Institut fuer Experimentelle Onkologie und Therapieforschung, Munich (Germany)

    2005-09-01

    Genes encoding for intracellular enzymes or transmembrane proteins are suitable as reporters, but may differ in terms of applicability for cardiac imaging. The aim of this study was to compare the human sodium iodide symporter gene (hNIS) with the herpes simplex virus type 1 thymidine kinase gene (HSV1-tk) as the reporter gene in non-invasive imaging of cardiac transgene expression with positron emission tomography (PET). Equal doses of adenoviral vectors encoding for hNIS, wild-type HSV1-tk, mutant HSV1-sr39tk or LacZ as the control gene were directly injected into the myocardium of 34 animals. Two days later, dynamic PET was performed with a clinical scanner, using reporter probes specific for the respective reporter gene. Imaging with {sup 13}N-ammonia was also performed to identify cardiac regions of interest. Kinetics differed significantly: {sup 124}I as the probe for hNIS showed rapid early uptake, remaining stable over time. Maximal myocardial concentration was 3.61{+-}1.15%. The nucleoside {sup 18}F-FHBG, as the specific probe for HSV1-sr39tk, showed increasing uptake over time, but maximal accumulation was significantly lower (1.45{+-}0.54%, P=0.0009). {sup 124}I-FIAU, as the specific probe for wild-type HSV1-tk, showed early uptake with subsequent washout. Maximal accumulation was lowest (0.63{+-}0.23%, P<0.0001). Post-mortem analysis by autoradiography and gamma counting confirmed the in vivo data. Reporter genes encoding for transporter proteins such as hNIS are an attractive alternative to overexpression of intracellular enzymes for cardiac gene product imaging. hNIS yielded higher signal intensity and imaging contrast for PET than did HSV1-tk and HSV1-sr39tk. Therefore, this approach may be preferable for the future monitoring of cardiac gene- or cell-based therapy. (orig.)

  8. Epidermal growth factor receptor tyrosine kinase (EGFR-TK) mutation testing in adults with locally advanced or metastatic non-small cell lung cancer: A systematic review and cost-effectiveness analysis

    OpenAIRE

    Westwood, Marie; Joore, Manuela; Whiting, Penny; Asselt, Thea; Ramaekers, Bram; Armstrong, Nigel; Misso, Kate; Severens, Hans; Kleijnen, Jos

    2014-01-01

    markdownabstract__Abstract__ Background: Non-small cell lung cancer (NSCLC) is the most common form of lung cancer. Some epidermal growth factor receptor tyrosine kinase (EGFR-TK) mutations make tumours responsive to treatment with EGFR-TK inhibitors (EGFR-TKIs) but less responsive to treatment with standard chemotherapy. Patients with NSCLC are therefore tested for EGFR-TK tumour gene mutations to inform treatment decisions. There are a variety of tests available to detect these mutations. T...

  9. Sensitivity of GC-EI/MS, GC-EI/MS/MS, LC-ESI/MS/MS, LC-Ag(+) CIS/MS/MS, and GC-ESI/MS/MS for analysis of anabolic steroids in doping control.

    Science.gov (United States)

    Cha, Eunju; Kim, Sohee; Kim, Ho Jun; Lee, Kang Mi; Kim, Ki Hun; Kwon, Oh-Seung; Lee, Jaeick

    2015-01-01

    This study compared the sensitivity of various separation and ionization methods, including gas chromatography with an electron ionization source (GC-EI), liquid chromatography with an electrospray ionization source (LC-ESI), and liquid chromatography with a silver ion coordination ion spray source (LC-Ag(+) CIS), coupled to a mass spectrometer (MS) for steroid analysis. Chromatographic conditions, mass spectrometric transitions, and ion source parameters were optimized. The majority of steroids in GC-EI/MS/MS and LC-Ag(+) CIS/MS/MS analysis showed higher sensitivities than those obtained with other analytical methods. The limits of detection (LODs) of 65 steroids by GC-EI/MS/MS, 68 steroids by LC-Ag(+) CIS/MS/MS, 56 steroids by GC-EI/MS, 54 steroids by LC-ESI/MS/MS, and 27 steroids by GC-ESI/MS/MS were below cut-off value of 2.0 ng/mL. LODs of steroids that formed protonated ions in LC-ESI/MS/MS analysis were all lower than the cut-off value. Several steroids such as unconjugated C3-hydroxyl with C17-hydroxyl structure showed higher sensitivities in GC-EI/MS/MS analysis relative to those obtained using the LC-based methods. The steroids containing 4, 9, 11-triene structures showed relatively poor sensitivities in GC-EI/MS and GC-ESI/MS/MS analysis. The results of this study provide information that may be useful for selecting suitable analytical methods for confirmatory analysis of steroids. Copyright © 2015 John Wiley & Sons, Ltd.

  10. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Viruses Clusters d Who Gets MS? Pediatric MS African Americans Hispanics & Latino/as d Multiple Sclerosis FAQs d ... Managing MS Resources for You and Your Practice Publications for Clinicians Publications for Your Patients MS Navigator ...

  11. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... to navigation Skip to content Menu Navigation National Multiple Sclerosis Society Sign In In Your Area Donate Donate ... of MS What Causes MS? Who Gets MS? Multiple Sclerosis FAQs Types of MS Related Conditions Symptoms & Diagnosis ...

  12. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Definition of MS Myelin Immune-Mediated Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters ... d Research News & Progress Research News MSPARIS2017 Stem Cells in MS Progressive MS Research Clinical Trials in ...

  13. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... MS d Diet, Exercise & Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another Condition Aging with MS Anesthesia and Surgery Managing Your MS ...

  14. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Gets MS? Pediatric MS African Americans Hispanics & Latino/as d Multiple Sclerosis FAQs d Types of MS ... Clinicians d Treating MS d Comprehensive Care Developing a Healthcare Team Make the Most of Your Doctor ...

  15. Axl and MerTK receptor tyrosine kinases maintain human macrophage efferocytic capacity in the presence of viral triggers.

    Science.gov (United States)

    Grabiec, Aleksander M; Goenka, Anu; Fife, Mark E; Fujimori, Toshifumi; Hussell, Tracy

    2018-02-05

    The requirement to remove apoptotic cells is equally important in homeostasis and inflammatory disease. In particular, during viral infections large quantities of infected cells undergo apoptosis and need to be efficiently cleared by phagocytes to prevent secondary necrosis. Although specific roles of several apoptotic cell sensors, such as the TAM (Tyro3, Axl, MerTK) receptor family, have been characterized in mouse models, little is known about their regulation and involvement in apoptotic cell uptake (efferocytosis) by human macrophages under inflammatory conditions. We show that whereas pro-inflammatory stimuli consistently downregulated MerTK expression in human monocyte-derived macrophages (MDMs), stimuli indicative of a viral infection, interferon-α (IFN-α) and the TLR3 ligand poly(I:C), specifically induced Axl expression and promoted binding of the bridging molecule Gas6. Axl induction by IFN-α and poly(I:C) was associated with higher MDM efferocytic capacity compared to cells treated with other pro-inflammatory stimuli, such as LPS and IFN-γ. While MerTK blocking antibody uniformly suppressed apoptotic cell uptake by MDMs, Axl blocking antibody significantly reduced efferocytosis by poly(I:C)-stimulated MDMs, but not by resting MDMs. Our observations demonstrate that Axl induction during viral infections contributes to maintaining macrophage capacity to engulf apoptotic cells, which may have important consequences for resolution of anti-viral immune responses. © 2018 The Authors. European Journal of Immunology published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. The Anti-Tumor Effects of Adipose Tissue Mesenchymal Stem Cell Transduced with HSV-Tk Gene on U-87-Driven Brain Tumor.

    Directory of Open Access Journals (Sweden)

    Suely Maymone de Melo

    Full Text Available Glioblastoma (GBM is an infiltrative tumor that is difficult to eradicate. Treating GBM with mesenchymal stem cells (MSCs that have been modified with the HSV-Tk suicide gene has brought significant advances mainly because MSCs are chemoattracted to GBM and kill tumor cells via a bystander effect. To use this strategy, abundantly present adipose-tissue-derived mesenchymal stem cells (AT-MSCs were evaluated for the treatment of GBM in mice. AT-MSCs were prepared using a mechanical protocol to avoid contamination with animal protein and transduced with HSV-Tk via a lentiviral vector. The U-87 glioblastoma cells cultured with AT-MSC-HSV-Tk died in the presence of 25 or 50 μM ganciclovir (GCV. U-87 glioblastoma cells injected into the brains of nude mice generated tumors larger than 3.5 mm2 after 4 weeks, but the injection of AT-MSC-HSV-Tk cells one week after the U-87 injection, combined with GCV treatment, drastically reduced tumors to smaller than 0.5 mm2. Immunohistochemical analysis of the tumors showed the presence of AT-MSC-HSV-Tk cells only within the tumor and its vicinity, but not in other areas of the brain, showing chemoattraction between them. The abundance of AT-MSCs and the easier to obtain them mechanically are strong advantages when compared to using MSCs from other tissues.

  17. Recombinant interleukin-2 enhanced the antitumor effect of ADV/RSV-HSV-tk/ACV therapy in a murine bladder cancer model.

    Science.gov (United States)

    Terao, Shuji; Shirakawa, Toshiro; Goda, Kazumasa; Kamidono, Sadao; Fujisawa, Masato; Gotoh, Akinobu

    2005-01-01

    Previous studies demonstrated the antitumor effects of IL-2 and ADV/RSV-HSV-tk in bladder tumor models. In our study, we employed the intramuscular injection of recombinant IL-2 combined with ADV/RSV-HSV-tk gene therapy in the MBT-2 murine bladder tumor model. In the in vitro study, after adenoviral gene transduction efficiency had been assessed, the cytotoxicity of ADV/RSV-HSV-tk/ACV was examined. In the in vivo study, ADV/RSV-HSV-tk was injected into MBT-2 subcutaneous tumors, ACV was injected intraperitoneally daily for 13 days and recombinant IL-2 was injected intramuscularly daily for 10 days. The X-gal staining of MBT-2 cells infected with 125 multiplicity of injection (MOI) indicated > 20% adenoviral gene transduction efficiency. The cell growth of MBT-2 infected with 125 MOI was significantly inhibited by 40 microM of ACV. In the in vivo study, the combination therapy significantly inhibited tumor growth in the MBT-2 tumor model. The systemic administration of recombinant IL-2 in combination with HSV-tk gene therapy exhibited an enhanced antitumor effect.

  18. Potent anti-tumor activity of telomerase-dependent and HSV-TK armed oncolytic adenovirus for non-small cell lung cancer in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    Ren Peng-Kang

    2010-05-01

    Full Text Available Abstract Background Non-small cell lung cancer (NSCLC is the leading cause of cancer related mortality, any improvements in therapeutic strategies are urgently required. In this study we generated a novel 'suicide gene' armed oncolytic adenoviral vector and investigated its antitumor effect both in vitro and in vivo. Methods Since the up-regulated expression of human telomerase reverse transcriptase (hTERT is a hallmark of alltypes of NSCLC, we chose hTERT promoter to transcriptionally control E1A gene expression to obtain adenoviral replication in NSCLC. In order to further enhance anti-tumor effect of this oncolytic adenoviral vector, we inserted a 'suicide gene' i.e. Herpes Simplex Virus Thymidine Kinase (HSV-TK into oncolytic adenoviral vector to engineer a novel armed oncolytic adenoviral vector 'Ad.hTERT-E1A-TK'. Results Ad.hTERT-E1A-TK efficiently killed different types of tumor cells including two types of NSCLC cells in vitro, causing no damage to normal primary fibroblasts. Furthermore, Ad.hTERT-E1A-TK infection combined with administration of prodrug gancyclovir (GCV resulted in more potent cytotoxicity on NSCLC cells, and synergistically suppressed human NSCLC tumor growth in nude mice. Conclusion The results from this study showed that Ad.hTERT-E1A-TK/GCV could be a potent but safe anti-tumor strategy for NSCLC biotherapy.

  19. Ms. Mentor Unmasked

    Science.gov (United States)

    Krebs, Paula

    2008-01-01

    This article presents an interview with Emily Toth, who writes the monthly "Ms. Mentor" academic advice column in the "Chronicle of Higher Education" and teaches in the English department at Louisiana State University, in Baton Rouge. She is the author of "Ms. Mentor's Impeccable Advice for Women in Academia" (1997), "Inside Peyton Place: The Life…

  20. Comparative study of cellular kinetics of reporter probe [{sup 131}I]FIAU in neonatal cardiac myocytes after transfer of HSV1-tk reporter gene with two vectors

    Energy Technology Data Exchange (ETDEWEB)

    Lan Xiaoli [Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China); Hubei Province Key Laboratory of Molecular Imaging, Wuhan 430022 (China)], E-mail: lxl730724@hotmail.com; Yin Xiaohua; Wang Ruihua; Liu Ying [Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China); Hubei Province Key Laboratory of Molecular Imaging, Wuhan 430022 (China); Zhang Yongxue [Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China) and Hubei Province Key Laboratory of Molecular Imaging, Wuhan 430022 (China)], E-mail: zhyx1229@163.com

    2009-02-15

    Aim: Reporter gene imaging is a promising approach for noninvasive monitoring of cardiac gene therapy. In this study, HSV1-tk (herpes simplex virus type 1 thymidine kinase) and FIAU (2'-fluoro-2'-deoxy-1-{beta}-D-arabinofuranosyl-5-iodouracil) were used as the reporter gene and probe, respectively. Cellular uptakes of radiolabeled FIAU of neonatal rat cardiac myocytes transferred with HSV1-tk were compared between two vectors, adenovirus and liposome. The aims of this study were to choose the better vector and to provide a theoretical basis for good nuclide images. Methods: Neonatal cardiac myocytes were obtained from rat heart by single collagenase digestion. HSV1-tk inserted into adenovirus vector (recombinant adenovirus type 5, Ad5-tk) and plasmid (pDC316-tk) coated with Lipofectamine 2000 (pDC316-tk/lipoplex) were developed; thus, HSV1-tk could be transferred into neonatal cardiac myocytes. FAU (2'-fluoro-2'-deoxy-1-{beta}-D-arabinofuranosyluracil) was labeled with {sup 131}I, and the product was assessed after purification with reversed-phase Sep-Pak C-18 column. The uptake rates of [{sup 131}I]FIAU in the transferred cardiac myocytes at different times (0.5, 1, 2, 3, 4 and 5 h) were detected. Furthermore, mRNA expression and protein expression of HSV1-tk were detected by semiquantitative reverse-transcriptase polymerase chain reaction and immunocytochemistry. Results: FAU could be labeled with {sup 131}I, and the labeling efficiency and radiochemical purity rates were 53.82{+-}2.05% and 94.85{+-}1.76%, respectively. Time-dependent increase of the accumulation of [{sup 131}I]FIAU was observed in both the Ad5-tk group and the pDC316/lipoplex group, and the highest uptake rate occurred at 5 h, with peak values of 12.55{+-}0.37% and 2.09{+-}0.34%, respectively. Greater uptakes of [{sup 131}I]FIAU in Ad5-tk-infected cells compared with pDC316/lipoplex-transfected ones occurred at all the time points (t=12.978-38.253, P<.01). The exogenous gene

  1. The targeted gene (KDRP-CD/TK) therapy of breast cancer mediated by SonoVue and ultrasound irradiation in vitro.

    Science.gov (United States)

    Li, Xing-hua; Zhou, Ping; Wang, Li-hua; Tian, Shuang-ming; Qian, Ying; Chen, Li-rong; Zhang, Ping

    2012-01-01

    Suicide gene therapy has become an effective therapy for breast cancer, and ultrasound targeted microbubble destruction (UTMD) has become a popular topic in the gene therapy field. In this study, MCF-7 cells with the KDR promoter and LSl74T cells without the KDR promoter were transfected with the recombinant plasmid pEGFP-KDRP-CD/TK using UTMD. The recombinant plasmid pEGFP-KDRP-CD/TK was transfected into MCF-7 and LS174T cells successfully with no significant difference in transfection efficiency (p>0.05). By RT-PCR, the CD/TK fusion gene was shown to be expressed in MCF-7 cells but not expressed in LS174T cells. In a cytotoxicity experiment, transgenic MCF-7 cells were sensitive to the prodrugs 5-FC and GCV. When both 5-FC and GCV were administered, the rate of cellular inhibition was significantly greater than that achieved when only one of the prodrugs was administered (p<0.001). Moreover, the inhibition rates achieved administering 5-FC, GCV and both 5-FC and GCV were all significantly greater than the gene transfection rate of 21.92±3.64% (p<0.001). However, transgenic LS174T cells were not sensitive to any prodrug. These results demonstrated that UTMD is a safe, effective and targeted gene delivery system. Also, the KDR promoter can drive expression of the CD/TK double suicide gene target in MCF-7 cells, and the targeted killing effect of the KDRP-CD/TK gene on MCF-7 cells in vitro has good synergy with expression of the CD/TK fusion gene. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. APPRECIATION AND CREATION IN CERAMIC ART LEARNING AS A FORM OF CULTURAL PRESERVATION FOR STUDENTS OF TK PANDEYAN 2 SUKOHARJO IN SURAKARTA RESIDENCY

    Directory of Open Access Journals (Sweden)

    Novita Wahyuningsih

    2017-03-01

    Full Text Available This study is aimed to describe the process of ceramic art learning process and to determine the impact of ceramic art learning for students of TK Pandeyan 2 Kindegarten of Sukoharjo by appreciating their creations. This method also intended to introduce and preserve ceramic culture for early age students. The method used is qualitative data analysis, in the form of data reduction, data presentation, and conclusion. The result shows that the ceramic art learning in TK Pandeyan 2 Kindegarten of Sukoharjo has positive effects as the teaching and learning process can be used as a medium for teachers and parents to understand children’s characters and personalities better.

  3. Activation of the HSV-TK promoter in control reporter vector pBLCAT5 by liganded nuclear retinoid receptor RXRα

    Directory of Open Access Journals (Sweden)

    Nikčević Gordana

    2006-01-01

    Full Text Available Widely used reporter vector systems for studying the putative regulatory DNA elements usually contain basal promoters from pathogenic mammalian viruses. It is a common assumption that reliable results can be achieved only if the viral promoter activity is unaffected by transacting factors or any experimental treatment. Here we report that liganded nuclear retinoid receptor RXRa stimulates the HSV-TK promoter in control reporter vector pBLCAT5. Thus, TK driven reporter vectors should be employed only after thorough testing of the regulation of this promoter under experimental stimuli for a particular research purpose in order to avoid unreliable interpretation of the assay results.

  4. IMS - MS Data Extractor

    Energy Technology Data Exchange (ETDEWEB)

    2015-10-20

    An automated drift time extraction and computed associated collision cross section software tool for small molecule analysis with ion mobility spectrometry-mass spectrometry (IMS-MS). The software automatically extracts drift times and computes associated collision cross sections for small molecules analyzed using ion mobility spectrometry-mass spectrometry (IMS-MS) based on a target list of expected ions provided by the user.

  5. Epidermal growth factor receptor tyrosine kinase (EGFR-TK) mutation testing in adults with locally advanced or metastatic non-small cell lung cancer : a systematic review and cost-effectiveness analysis

    NARCIS (Netherlands)

    Westwood, Marie; Joore, Manuela; Whiting, Penny; van Asselt, Thea; Ramaekers, Bram; Armstrong, Nigel; Misso, Kate; Severens, Johan; Kleijnen, Jos

    BACKGROUND: Non-small cell lung cancer (NSCLC) is the most common form of lung cancer. Some epidermal growth factor receptor tyrosine kinase (EGFR-TK) mutations make tumours responsive to treatment with EGFR-TK inhibitors (EGFR-TKIs) but less responsive to treatment with standard chemotherapy.

  6. Epidermal growth factor receptor tyrosine kinase (EGFR-TK) mutation testing in adults with locally advanced or metastatic non-small cell lung cancer: A systematic review and cost-effectiveness analysis

    NARCIS (Netherlands)

    M. Westwood (Marie); M.A. Joore (Manuela); P. Whiting (Penny); T. van Asselt (Thea); B.L.T. Ramaekers (Bram); N. Armstrong (Nigel); K. Misso (Kate); J.L. Severens (Hans); J. Kleijnen (Jos)

    2014-01-01

    markdownabstract__Abstract__ Background: Non-small cell lung cancer (NSCLC) is the most common form of lung cancer. Some epidermal growth factor receptor tyrosine kinase (EGFR-TK) mutations make tumours responsive to treatment with EGFR-TK inhibitors (EGFR-TKIs) but less responsive to treatment

  7. TK-Team AS on üllatavalt edukas firma ja töökale töötajale hea tööandja / Raivo Feldmann

    Index Scriptorium Estoniae

    Feldmann, Raivo

    2007-01-01

    TK Team AS on Taulukeskus TK-Team AB kontserni ettevõte, kümne tegevusaasta jooksul on ta läbi teinud märkimisväärse arengu, mis võimaldab firmal edukalt konkureerida ka maailma tahvliturul. Tegevjuht Ants Reinumägi tutvustab ettevõtet

  8. Enhanced therapeutic effect of multiple injections of HSV-TK + GCV gene therapy in combination with ionizing radiation in a mouse mammary tumor model

    International Nuclear Information System (INIS)

    Vlachaki, Maria T.; Chhikara, Madhu; Aguilar, Laura; Zhu Xiaohong; Chiu, Kam J.; Woo, Shiao; Teh, Bin S.; Thompson, Timothy C.; Butler, E. Brian; Aguilar-Cordova, Estuardo

    2001-01-01

    Purpose: Standard therapies for breast cancer lack tumor specificity and have significant risk for recurrence and toxicities. Herpes simplex virus-thymidine kinase (HSV-tk) gene therapy combined with radiation therapy (XRT) may be effective because of complementary mechanisms and distinct toxicity profiles. HSV-tk gene therapy followed by systemic administration of ganciclovir (GCV) enhances radiation-induced DNA damage by generating high local concentrations of phosphorylated nucleotide analogs that increase radiation-induced DNA breaks and interfere with DNA repair mechanisms. In addition, radiation-induced membrane damage enhances the 'bystander effect' by facilitating transfer of nucleotide analogs to neighboring nontransduced cells and by promoting local and systemic immune responses. This study assesses the effect of single and multiple courses of HSV-tk gene therapy in combination with ionizing radiation in a mouse mammary cancer model. Methods and Materials: Mouse mammary TM40D tumors transplanted s.c. in syngeneic immunocompetent BALB-c mice were treated with either adenoviral-mediated HSV-tk gene therapy or local radiation or the combination of gene and radiation therapy. A vector consisting of a replication-deficient (E1-deleted) adenovirus type 5 was injected intratumorally to administer the HSV-tk gene, and GCV was initiated 24 h later for a total of 6 days. Radiation was given as a single dose of 5 Gy 48 h after the HSV-tk injection. A metastatic model was developed by tail vein injection of TM40D cells on the same day that the s.c. tumors were established. Systemic antitumor effect was evaluated by counting the number of lung nodules after treating only the primary tumors with gene therapy, radiation, or the combination of gene and radiation therapy. To assess the therapeutic efficacy of multiple courses of this combinatorial approach, one, two, and three courses of HSV-tk + GCV gene therapy, in combination with radiation, were compared to HSV-tk or

  9. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters d Who Gets MS? Pediatric MS ... bowel and bladder problems, and problems with cognition (learning and memory or information processing). People with progressive ...

  10. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters d Who Gets MS? Pediatric MS ... times as often as men; in PPMS, the number of women and men are approximately equal. RRMS ...

  11. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters d Who Gets MS? Pediatric MS ... Well-Being Mood Changes d Spiritual Well-Being Building Spirituality into Your Life d Cognitive Health d ...

  12. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... without symptoms of which the person is aware. What happens in RRMS? Relapsing-remitting MS is defined ... Professionals Mental Health Professionals Health and Wellness Professionals What Is MS? Symptoms & Diagnosis Treating MS Resources & Support ...

  13. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Types of MS Relapsing-remitting MS (RRMS) Share this page Facebook Twitter Email Relapsing-remitting MS (RRMS) ... been living with RRMS for at least 10 years. The most common symptoms reported in RRMS include ...

  14. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Trials in MS Wellness and Lifestyle Research Diet Vitamin D How and Why Do Scientists Share Results ... The National MS Society is Here to Help Need More Information? We Are Here Our MS Navigators ...

  15. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Relapsing-remitting MS (RRMS) Share this page Facebook Twitter Email Relapsing-remitting MS (RRMS) Relapsing-remitting MS (RRMS) Diagnosing RRMS Treating RRMS Research in RRMS Share Smaller Text Larger Text Print ...

  16. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Viruses Clusters d Who Gets MS? Pediatric MS African Americans Hispanics & Latino/as d Multiple Sclerosis FAQs d ... Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another Condition ...

  17. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Disease T Cells d What Causes MS? Disproved Theories Viruses Clusters d Who Gets MS? Pediatric MS ... of Distinction Lawry Circle Circle of Influence d Planned Giving d Other Ways to Give Donate by ...

  18. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Room Events at a Glance MS the Disease Public Service Announcements In the News Archives d MS ... For Professionals Researchers Physicians Nurses Rehabilitation Professionals Mental Health Professionals Health and Wellness Professionals What Is MS? ...

  19. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Disproved Theories Viruses Clusters d Who Gets MS? Pediatric MS African Americans Hispanics & Latino/as d Multiple ... Questions to Ask d Resources for Specific Populations Pediatric MS Support Veterans with Multiple Sclerosis d Find ...

  20. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another Condition Aging with MS ...

  1. Influence of p53 status on the HSV-Tk/GCV-induced bystander effect in a panel of human ovarian carcinoma cell lines

    NARCIS (Netherlands)

    van Dillen, IJ; Mulder, NH; Sluiter, WJ; Meijer, C; de Jong, S; Loncarek, J; Mesnil, M; de Vries, EFJ; Vaalburg, W; Hospers, GAP

    2005-01-01

    The p53 protein is mutated in 50% of all human tumors and plays a key role in apoptosis, cell cycle, and the expression of several genes. We investigated if p53 mutations influence the herpes simplex virus thymidine kinase (HSV-tk)/ganciclovir (GCV)-induced bystander effect (BE). Additionally, we

  2. Noninvasive theranostic imaging of HSV-TK/GCV suicide gene therapy in liver cancer by folate-targeted quantum dot-based liposomes

    NARCIS (Netherlands)

    Shao, D.; Li, J.; Pan, Y.; Zhang, X.; Zheng, X.; Wang, Z.; Zhang, M.; Zhang, H.; Chen, L.

    2015-01-01

    Theranostics is emerging as a popular strategy for cancer therapy; thanks to the development of nano-technology. In this work, we have combined an HSV-TK/GCV suicide gene system and near-infrared quantum dots, as the former is quite effective in liver cancer treatment and the latter facilitates

  3. City vs. climate changes - the future and its (unsustainability: The global applicability of Sarriguren and Jätkäsaari?

    Directory of Open Access Journals (Sweden)

    Stupar Aleksandra

    2011-01-01

    Full Text Available The global reality, intensively exposed to the challenges caused by climate changes, demands new ideas, methods and tools which could eventually prevent or mitigate the consequences of carbon emissions. Therefore, the contemporary cities, as focal points of the new global comprehension, have become testing grounds for numerous initiatives based upon imperatives of sustainability, environmental consciousness, energy efficiency and ‘green’ life style. In Serbia, experience in matters of the built environment has had a tendency to be formed exclusively on the basis of authentic local initiatives, aspirations, successes and failures. In the global effort for sustainable development, considering the commitments imposed by climate change and carbon emission, a greater reliance on results achieved elsewhere would be a welcome change. Consequently, this article will present and comment two eco-driven projects - Sarriguren and Jätkäsaari, both covering a wide range of useful ideas, intriguing concepts and globally applicable solutions for the challenges caused by climate shifts.

  4. Purification and Characterization of the Principal Antimutagenic Bioactive as Ethoxy-Substituted Phylloquinone from Spinach (Spinacea oleracea L.) Based on Evaluation in Models Including Human Lymphoblast TK+/-Cells.

    Science.gov (United States)

    Kumar, Sanjeev; Chatterjee, Suchandra; Tripathi, Jyoti; Gautam, Satyendra

    2016-11-23

    During in vitro analysis, spinach (Spinacea oleracea L.) leaf extracts displayed varying antimutagenicity when analyzed in models including human lymphoblast (TK +/- ) cell line (thymidine kinase gene mutation assay) and Escherichia coli MG1655 (rifampicin resistance assay) against chemically (ethyl methanesulfonate and 5-azacytidine) induced mutagenicity. Highest antimutagenicity was displayed by the quinone extract. The principal bioactive compound exhibited fluorescence in TLC at 366 nm (termed C4) resolved at R f 0.32 and t R 15.2 min in TLC and HPLC, respectively. On the TLC plate, three spots (C1-C3), observed at 254 nm, displayed comparatively lesser antimutagenicity. Furthermore, biochemical and spectroscopic analyses using MALDI-TOF MS and NMR indicated the nature of the potent compound (C4) as an ethoxy-substituted phylloquinone derivative [2-ethoxy-3-((E)-3,7,11,15-tetramethylhexadec-2-enyl)naphthalene-1,4-dione]. The C4 compound did not display any cytotoxicity and hence possesses significant nutraceutical-based intervention possibility to combat the onset of mutation-associated disease(s).

  5. A combination hepatoma-targeted therapy based on nanotechnology: pHRE-Egr1-HSV-TK/131I-antiAFPMcAb-GCV/MFH

    Science.gov (United States)

    Lin, Mei; Huang, Junxing; Jiang, Xingmao; Zhang, Jia; Yu, Hong; Ye, Jun; Zhang, Dongsheng

    2016-09-01

    Combination targeted therapy is a promising cancer therapeutic strategy. Here, using PEI-Mn0.5Zn0.5Fe2O4 nanoparticles (PEI-MZF-NPs) as magnetic media for MFH (magnetic fluid hyperthermia) and gene transfer vector for gene-therapy, a combined therapy, pHRE-Egr1-HSV-TK/131I-antiAFPMcAb-GCV/MFH, for hepatoma is developed. AntiAFPMcAb (Monoclonal antibody AFP) is exploited for targeting. The plasmids pHRE-Egr1-HSV-TK are achieved by incorporation of pEgr1-HSV-TK and pHRE-Egr1-EGFP. Restriction enzyme digestion and PCR confirm the recombinant plasmids pHRE-Egr1-HSV-TK are successfully constructed. After exposure to the magnetic field, PEI-MZF-NPs/pHRE-Egr1-EGFP fluid is warmed rapidly and then the temperature is maintained at 43 °C or so, which is quite appropriate for cancer treatment. The gene expression reaches the peak when treated with 200 μCi 131I for 24 hours, indicating that the dose of 200 μCi might be the optimal dose for irradiation and 24 h irradiation later is the best time to initiate MFH. The in vitro and in vivo experiments demonstrate that pHRE-Egr1-HSV-TK/131I-antiAFPMcAb-GCV/MFH can greatly suppress hepatic tumor cell proliferation and induce cell apoptosis and necrosis and effectively inhibit the tumor growth, much better than any monotherapy does alone. Furthermore, the combination therapy has few or no adverse effects. It might be applicable as a strategy to treat hepatic cancer.

  6. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... the Challenges of MS You CAN! Webcasts DVDs Books For Kids: Keep S'myelin Información en Español Brochures ... Managing MS and Another Condition Aging with MS Anesthesia and Surgery Managing Your MS d Emotional Well- ...

  7. Determination of Glyphosate Levels in Breast Milk Samples from Germany by LC-MS/MS and GC-MS/MS

    NARCIS (Netherlands)

    Steinborn, Angelika; Alder, Lutz; Michalski, Britta; Zomer, Paul; Bendig, Paul; Martinez, Sandra Aleson; Mol, Hans G.J.; Class, Thomas J.; Costa Pinheiro, Nathalie

    2016-01-01

    This study describes the validation and application of two independent analytical methods for the determination of glyphosate in breast milk. They are based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS), respectively. For

  8. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... MS Relapsing-remitting MS (RRMS) Share this page Facebook Twitter Email Relapsing-remitting MS (RRMS) Relapsing-remitting ... Here Start Here Colophon Stay Informed Join Us Facebook Twitter LinkedIn YouTube Pinterest MS Connection About the ...

  9. Human herpesviruses and MS

    DEFF Research Database (Denmark)

    Christensen, Tove

    2007-01-01

    Environmental factors operate on a background of genetic susceptibility in MS pathogenesis; the human herpesviruses (HHV) are likely candidates for such factors. HHV share a number of properties: they are almost ubiquitous, they are highly prevalent worldwide, they all cause latent infections...

  10. ICP-MS Workshop

    Energy Technology Data Exchange (ETDEWEB)

    Carman, April J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Eiden, Gregory C. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2014-11-01

    This is a short document that explains the materials that will be transmitted to LLNL and DNN HQ regarding the ICP-MS Workshop held at PNNL June 17-19th. The goal of the information is to pass on to LLNL information regarding the planning and preparations for the Workshop at PNNL in preparation of the SIMS workshop at LLNL.

  11. Long term follow up of patients after allogeneic stem cell transplantation and transfusion of HSV-Tk transduced T-cells.

    Directory of Open Access Journals (Sweden)

    Eva Maria Weissinger

    2015-04-01

    Full Text Available Allogeneic stem cell transplantation (allo-HSCT is one of the curative treatments for hematologic malignancies, but is hampered by severe complications, such as acute or chronic graft-versus-host-disease (aGvHD; cGvHD and infections. CD34-selcetion of stem cells reduces the risk of aGvHD, but also leads to increased infectious complications and relapse. Thus, we studied the efficacy, safety and feasibility of transfer of gene modified donor T-cells shortly after allo-HSCT in two clinical trials between 2002 and 2007 and here we compare the results to unmodified donor leukocyte transfusion (DLI. The aim of these trials was to provide patients with the protection of T-cells after T-cell-depleted allo-HSCT in the matched or mismatched donor setting with an option to delete transduced T-cells, if severe aGvHD occurred within the trial period. Donor-T-cells were transduced with the replication-deficient retrovirus SFCMM-3, expressing HSV-Tk and the truncated LNGFR for selection of transduced cells. Transduced cells were transfused either after day +60 (matched donors or on day +42 (haploidentical donors.Nine patients were included in the first trial (MHH; 2002 until 2007 2 were included in TK007 (2005-2009 and 6 serve as a control group for outcome after haploidentical transplantation without HSV-TK-transduced DLI. Three patients developed acute GvHD, two had grade I of the skin, one had aGvHD on day +131 (post-HSCT; +89 post-HSV-Tk DLI grade II, which was successfully controlled by ganciclovir (GCV. Donor chimerism was stabilized after transfusion of the transduced cells in all patients treated. Functionality of HSV-Tk gene expressing T-cells was shown by loss of bcr-abl gene expression as well as by control of cytomegalovirus-reactivation. To date, 6patients have relapsed and died, 2 after a second HSCT without T-cell depletion or administration of unmodified T-cells. Eleven patients (7 post-HSV-Tk DLI are alive and well to date.

  12. Analysis of cellular response by exposure to acute or chronic radiation in human lymphoblastoid TK-6 cells

    Science.gov (United States)

    Ohnishi, T.; Yasumoto, J.; Takahashi, A.; Ohnishi, K.

    To clarify the biological effects of low-dose rate radiation on human health for long-term stay in space, we analyzed the induction of apoptosis and apoptosis-related gene expression after irradiation with different dose-rate in human lymphoblastoid TK-6 cells harboring wild-type p53 gene. We irradiated TK-6 cells by X-ray at 1.5 Gy (1 Gy/min) and then sampled at 25 hr after culturing. We also irradiated by gamma-ray at 1.5 Gy (1 mGy/min) and then sampled immediately or 25 hr after irradiation. For DNA ladder analysis, we extracted DNA from these samples and electrophoresed with 2% agarose gel. In addition, we extracted mRNA from these samples for DNA-array analysis. mRNA from non-irradiated cells was used as a control. After labeling the cDNA against mRNA with [α -33P]-dCTP and hybridizing onto DNA array (Human Apoptosis Expression Array, R&D Systems), we scanned the profiles of the spots by a phosphorimager (BAS5000, FUJI FILM) and calculated using a NIH Image program. The data of each DNA-array were normalized with eight kinds of house keeping genes. We analyzed the expression level of apoptosis-related genes such as p53-related, Bcl-2 family, Caspase family and Fas-related genes. DNA ladders were obviously detected in the cells exposed to a high dose-rate radiation. We detected the induction of the gene expression of apoptosis-promotive genes. In contrast, almost no apoptosis was observed in the cells exposed to the chronic radiation at a low dose-rate. In addition, we detected the induction of the gene expression of apoptosis-suppressive genes as compared with apoptosis promotive-genes immediately after chronic irradiation. These results lead the importance of biological meaning of exposure to radiation at low dose-rate from an aspect of carcinogenesis. Finally, the effects of chronic irradiation become a highly important issue in space radiation biology for human health.

  13. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... working effectively. How does RRMS differ from progressive types of MS? While RRMS is defined by attacks ... Brochure Working with MS (.pdf) Download Brochure Taming Stress (.pdf) Download Brochure Multiple Sclerosis and Your Emotions (. ...

  14. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Treating MS Comprehensive Care Find an MS Care Provider Medications Managing Relapses Rehabilitation Complementary & Alternative Medicines For Clinicians Resources & Support Library & Education Programs Find Support Advanced Care ...

  15. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... the News Archives d MS Prevalence d Charitable Ratings d Corporate Support and Programs d Helpful Links ... Values Financials News Press Room MS Prevalence Charitable Ratings Corporate Support Helpful Links Donate Society Store MSConnection ...

  16. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... d Spiritual Well-Being Building Spirituality into Your Life d Cognitive Health d Work, Home & Leisure Employment ... Partnership Opportunities d Personal Stories Ambassadors & Familiar Faces Life with MS Givers Shakers Families with MS Seekers ...

  17. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Contact Us Newly Diagnosed If you or somone close to you has recently been diagnosed, access our MS information and resources. Start ... Room MS Prevalence Charitable Ratings Corporate Support Helpful Links ...

  18. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... d Research We Fund Stopping MS In Its Tracks Restoring What's Been Lost Ending the Disease Forever ... which produce the symptoms of MS . Because the location of the damage is so variable, no two ...

  19. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... of Programs and Events Living Well with MS Diet, Exercise & Healthy Behaviors Emotional Well-Being Spiritual Well- ... and Events d Living Well with MS d Diet, Exercise & Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature ...

  20. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Your Practice Publications for Clinicians Publications for Your Patients MS Navigator Program Programs and Services for Your Patients Contact Us Clinical Fellows d Careers in MS ...

  1. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Programs and Events Living Well with MS Diet, Exercise & Healthy Behaviors Emotional Well-Being Spiritual Well-Being ... Events d Living Well with MS d Diet, Exercise & Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity ...

  2. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... progressive MS (SPMS) d Related Conditions Acute Disseminated Encephalomyelitis (ADEM) Balo’s Disease HTLV-I Associated Myelopathy (HAM) ... Out Lyme Disease Lupus Neuromyelitis Optica Acute Disseminated Encephalomyelitis (ADEM) d For Clinicians d Treating MS d ...

  3. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... is working effectively. How does RRMS differ from progressive types of MS? While RRMS is defined by attacks of inflammation (relapses) in the CNS, progressive forms of MS involve much less of this ...

  4. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Banks d Research News & Progress Research News MSPARIS2017 Stem Cells in MS Progressive MS Research Clinical Trials ... inflammation. People with RRMS tend to have more brain lesions — also called plaques or scars — on magnetic ...

  5. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Team Make the Most of Your Doctor Visits Advance Medical Directives d Find an MS Care Provider ... Research Diet Vitamin D How and Why Do Scientists Share Results d Research We Fund Stopping MS ...

  6. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... bowel and bladder problems, and problems with cognition (learning and memory or information processing). People with progressive ... they may have. Diagnosing relapsing-remitting MS (RRMS) Learn More Learn More Treating relapsing-remitting MS (RRMS) ...

  7. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Events Living Well with MS Diet, Exercise & Healthy Behaviors Emotional Well-Being Spiritual Well-Being Cognitive Health ... Living Well with MS d Diet, Exercise & Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations ...

  8. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... FES) d Complementary & Alternative Medicines Chiropractic Therapy Marijuana Massage and Bodywork Acupuncture d For Clinicians d Resources & ... Teams Partnership Opportunities d Personal Stories Ambassadors & Familiar Faces Life with MS Givers Shakers Families with MS ...

  9. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another ... working effectively. How does RRMS differ from progressive types of MS? While RRMS is defined by attacks ...

  10. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... MS? While RRMS is defined by attacks of inflammation (relapses) in the CNS, progressive forms of MS involve much less of this type of inflammation. People with RRMS tend to have more brain ...

  11. Reversible tetramerization of human TK1 to the high catalytic efficient form is induced by pyrophosphate, in addition to tripolyphosphates, or high enzyme concentration

    DEFF Research Database (Denmark)

    Munch-Petersen, Birgitte

    2009-01-01

    of ATP is necessary for tetramerisation and how the reaction velocity is influenced by the enzyme concentration. The results show that only two or three of the phosphate groups of ATP are necessary for tetramerisation, and that kinetics and tetramerisation are closely related. Furthermore, enzyme...... concentration was found to have a pivotal effect on catalytic efficiency.......Thymidine kinase (TK1) is a key enzyme in the salvage pathway of deoxyribonucleotide metabolism catalyzing the first step in the synthesis of dTTP by the transfer of a gamma-phosphate group from a nucleoside triphosphate to the 5´-hydroxyl group of thymidine forming dTMP. Human TK1 is cytosolic...

  12. Imaging grafted cells with [18F]FHBG using an optimized HSV1-TK mammalian expression vector in a brain injury rodent model.

    Directory of Open Access Journals (Sweden)

    Anne-Sophie Salabert

    Full Text Available Cell transplantation is an innovative therapeutic approach after brain injury to compensate for tissue damage. To have real-time longitudinal monitoring of intracerebrally grafted cells, we explored the feasibility of a molecular imaging approach using thymidine kinase HSV1-TK gene encoding and [18F]FHBG as a reporter probe to image enzyme expression.A stable neuronal cell line expressing HSV1-TK was developed with an optimised mammalian expression vector to ensure long-term transgene expression. After [18F]FHBG incubation under defined parameters, calibration ranges from 1 X 104 to 3 X 106 Neuro2A-TK cells were analysed by gamma counter or by PET-camera. In parallel, grafting with different quantities of [18F]FHBG prelabelled Neuro2A-TK cells was carried out in a rat brain injury model induced by stereotaxic injection of malonate toxin. Image acquisition of the rats was then performed with PET/CT camera to study the [18F]FHBG signal of transplanted cells in vivo.Under the optimised incubation conditions, [18F]FHBG cell uptake rate was around 2.52%. In-vitro calibration range analysis shows a clear linear correlation between the number of cells and the signal intensity. The PET signal emitted into rat brain correlated well with the number of cells injected and the number of surviving grafted cells was recorded via the in-vitro calibration range. PET/CT acquisitions also allowed validation of the stereotaxic injection procedure. Technique sensitivity was evaluated under 5 X 104 grafted cells in vivo. No [18F]FHBG or [18F]metabolite release was observed showing a stable cell uptake even 2 h post-graft.The development of this kind of approach will allow grafting to be controlled and ensure longitudinal follow-up of cell viability and biodistribution after intracerebral injection.

  13. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... and problems with cognition (learning and memory or information processing). People with progressive forms of MS are more likely to experience gradually worsening problems with walking and mobility, along with whatever other symptoms they may have. Diagnosing ... MS Society is Here to Help Need More Information? We Are Here Our MS Navigators help identify ...

  14. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Living Well with MS Diet, Exercise & Healthy Behaviors Emotional Well-Being Spiritual Well-Being Cognitive Health Work, ... MS Anesthesia and Surgery Managing Your MS d Emotional Well-Being Mood Changes d Spiritual Well-Being ...

  15. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another Condition Aging with MS Anesthesia and Surgery Managing Your MS d Emotional Well-Being Mood Changes d Spiritual Well-Being Building Spirituality into Your ...

  16. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... MS? d Definition of MS Myelin Immune-Mediated Disease T Cells d What Causes MS? Disproved Theories ... d Related Conditions Acute Disseminated Encephalomyelitis (ADEM) Balo’s Disease HTLV-I Associated Myelopathy (HAM) Neuromyelitis Optica (NMO) ...

  17. Safety and immunogenicity of a gE/gI/TK gene-deleted pseudorabies virus variant expressing the E2 protein of classical swine fever virus in pigs.

    Science.gov (United States)

    Lei, Jian-Lin; Xia, Shui-Li; Wang, Yimin; Du, Mingliang; Xiang, Guang-Tao; Cong, Xin; Luo, Yuzi; Li, Lian-Feng; Zhang, Lingkai; Yu, Jiahui; Hu, Yonghao; Qiu, Hua-Ji; Sun, Yuan

    2016-06-01

    Classical swine fever (CSF) and pseudorabies (PR) are both major infectious diseases of pigs, causing enormous economic losses to the swine industry in many countries. A marker vaccine that enables differentiation of infected from vaccinated animals (DIVA) is highly desirable for control and eradication of these two diseases in endemic areas. Since late 2011, PR outbreaks have been frequently reported in many Bartha-K61-vaccinated pig farms in China. It has been demonstrated that a pseudorabies virus (PRV) variant with altered antigenicity and increased pathogenicity was responsible for the outbreaks. Previously, we showed that rPRVTJ-delgE/gI/TK, a gE/gI/TK-deleted PRV variant, was safe for susceptible animals and provided a complete protection against lethal PRV variant challenge, indicating that rPRVTJ-delgE/gI/TK can be used as an attractive vaccine vector. To develop a safe bivalent vaccine against CSF and PR, we generated a recombinant virus rPRVTJ-delgE/gI/TK-E2 expressing the E2 protein of classical swine fever virus (CSFV) based on rPRVTJ-delgE/gI/TK and evaluated its safety and immunogenicity in pigs. The results indicated that pigs (n=5) immunized with rPRVTJ-delgE/gI/TK-E2 of different doses did not exhibit clinical signs or viral shedding following immunization, the immunized pigs produced anti-PRV or anti-CSFV neutralizing antibodies and the pigs immunized with 10(6) or 10(5) TCID50 rPRVTJ-delgE/gI/TK-E2 were completely protected against the lethal challenge with either CSFV Shimen strain or variant PRV TJ strain. These findings suggest that rPRVTJ-delgE/gI/TK-E2 is a promising bivalent DIVA vaccine candidate against CSFV and PRV coinfections. Copyright © 2016 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

  18. Low doses of gamma ionizing radiation increase hprt mutant frequencies of TK6 cells without triggering the mutator phenotype pathway

    Energy Technology Data Exchange (ETDEWEB)

    Ayres, Flavio Monteiro; Glickman, Barry W.; Steele, Patricia [University of Victoria, BC (Canada). Dept. of Biology. Centre for Environmental; Cruz, Aparecido Divino da [Universidade Estadual de Goias, Anapolis, GO (Brazil). Dept. de Biologia. Nucleo de Pesquisas Replicon]. E-mail: acruz@ucg.br

    2006-07-01

    The TK6 lymphoblastoid cell line is known to be mismatch repair (MMR) and p53 proficient. Deficiency in MMR results in a mutator phenotype characterized by microsatellite instability (MSI) and increased hprt mutant frequency (MF). Increased hprt MF is also a biomarker of effect for exposure to ionizing radiation. In order to test if a mutator phenotype could be induced by low doses of gamma ionizing radiation, an hprt cloning assay and a MSI investigation were performed after radiation exposure. The spontaneous MF was 1.6 x 10-6. The groups exposed to 0.2, 0.5 and 1.0 Gy had hprt MFs of 2.3, 3.3 and 2.2 x 10-6, respectively. The spontaneous MSI frequency per allele in non-selected cells was 5.4 x 10-3, as evidenced at the loci D11S35, nm23-H1, D8S135 and p53. MSI frequencies in the groups exposed to 0.2, 0.5 and 1.0 Gy were found to be < 4.7, < 7.7 and < 12 x 10-3, respectively. The frequencies of hprt mutants and MSI found in this study suggest that low doses of ionizing radiation increase hprt mutant frequency without triggering the mutator phenotype pathway. (author)

  19. LC-MS (/MS) in clinical toxicology screening methods.

    Science.gov (United States)

    Viette, Véronique; Hochstrasser, Denis; Fathi, Marc

    2012-01-01

    Toxicological screening is the analysis of biological samples to detect and identify unknown compounds. The high selectivity and sensitivity of liquid chromatography (LC) coupled to mass spectrometry (MS) or tandem mass spectrometry (MS/MS) technology provide an attractive alternative to the current methods (LC-UV, GC/MS, etc.). For these reasons, an increasing number of applications are being published. This paper is a brief overview of LC-MS(/MS) screening methods developed for clinical toxicology in recent years. Various sample treatments, chromatographic separations and detection by mass spectrometry can be combined to obtain screening methods adapted to the constraints and needs of clinical toxicology laboratories. Currently the techniques are in the hands of specialists, mainly in academic institutions. However, the evolution in technology should allow application of these techniques as a tool in toxicology laboratories, thus allowing a more widespread exploitation of their potential.

  20. Decrease in protein binding and its effect on toxicokinetics (TK)/toxicodynamics (TD) of diclofenac and propranolol in pregnant rats.

    Science.gov (United States)

    Miida, Hiroaki; Noritake, Yumiko; Shimoda, Hitomi; Honda, Kumi; Matsuoka, Toshiki; Sakurai, Ken; Shirai, Makoto; Manabe, Sunao; Takasaki, Wataru; Ueno, Koichi

    2008-12-01

    Plasma protein binding is an important factor for the kinetics of drugs and how they act since it is the first step in drug distribution. Physiological changes in pregnancy, which include plasma composition, can affect drug binding and subsequent drug response. In the present study, we investigated the toxicokinetics (TK) and/or toxicodynamics (TD) of diclofenac and propranolol comparing pregnant Sparague-Dawley (SD) rats with non-pregnant SD rats in terms of protein binding and drug distribution. Diclofenac and propranolol are reported to bind to albumin and alpha(1)-acid glycoprotein (AGP), respectively. After a single administration of diclofenac, the area under plasma concentration-time curve (AUC) based on free diclofenac in pregnant rats was 3.9 times higher than that in non-pregnant rats. This difference is considered to be due to a lower concentration of serum albumin and a higher concentration of non-esterified fatty acid (NEFA) that inhibits drug binding to albumin, in pregnant rats. In histopathological examination, more severe gastrointestinal toxicity was observed in pregnant rats at 24 hr after dosing. This severe toxicity was likely to be correlated with the higher AUC. With respect to propranolol, the difference of the AUC based on free propranolol was not clear although the concentration of serum AGP was lower in pregnant rats. However, the binding analysis data suggested a difference of protein binding at a lower propranolol concentration range. Consequently, lowered serum proteins and increased NEFA in pregnant rats can lead to low protein binding, subsequent increase in free drug concentrations, and eventual increase in the TD of drugs.

  1. Pengembangan Kemampuan Kognitif Dan Sosial-Emosional Melalui Penerapan Media Balok Dan Bermain Peran Pada Siswa Tk Kuntum Mekar, Lampung

    Directory of Open Access Journals (Sweden)

    Leli Fertiliana Dea

    2017-12-01

    Full Text Available This study aims to (1 to determine the use of block media in the cognitive and social development of children in Kuntum Mekar 2 Bandar Lampung, (2 to know the application of bemain role method in developing cognitive and social emotional of children in Kuntum Mekar Kindergarten 2 Bandar Lampung, (3 to know the successful application of block media and role playing method in early childhood cognitive and social development. This research uses qualitative research method, the data obtained through observation and interview, then the data is analyzed by data reduction, data presentation and concluded. From the research, it can be concluded that: First, the application of beam media in Kuntum Mekar TK 2 Bandar Lampung succeeded. Use the following steps: (a The teacher prepares the tools. (b The teacher accepts the children present. (c Master explains how to play blocks. (d The child builds with the beam and the teacher is just watching. Second, the application of role playing in Kuntum Mekar 2 Bandar Lampung was successful. Apply using the following steps: (a Teachers collect children for direction and rules in the game, (b Teacher discusses the tools to be used, (c Teacher gives assignments to the child. Third, the successful application of beam media and role playing methods in early childhood cognitive and social development is evidenced by children developing as expected, as children are able to recognize objects by function, the child is able to classify objects by shape, color and size, the child is able to recognize the ABC pattern , children are able to recognize the concept of numbers, children are able to say a lot of objects 1-10 and children are able to cooperate with friends, children are able to know etiquette and courtesy in accordance with local social cultural values, children are able to understand the rules and discipline and children are able to show empathy attitude.

  2. Study of [18F]FLT and [123I]IaraU for cellular imaging in HSV1 tk-transfected murine fibrosarcoma cells: evaluation of the tracer uptake using 5-fluoro, 5-iodo and 5-iodovinyl arabinosyl uridines as competitive probes.

    Science.gov (United States)

    Huang, Ho-Lien; Chiang, Li-Wu; Chen, Jia-Rong; Yang, Wen K; Jeng, Kee-Ching; Chen, Jenn-Tzong; Duh, Ting-Shien; Lin, Wuu-Jyh; Farn, Shiou-Shiow; Chiang, Chi-Shiun; Huang, Chia-Wen; Lin, Kun-I; Yu, Chung-Shan

    2012-04-01

    As one of the most intensively studied probes for imaging of the cellular proliferation, [(18)F]FLT was investigated whether the targeting specificity of thymidine kinase 1 (TK1) dependency could be enhanced through a synergistic effect mediated by herpes simplex type 1 virus (HSV1) tk gene in terms of the TK1 or TK2 expression. 5-[(123)I]Iodo arabinosyl uridine ([(123)I]IaraU) was prepared in a radiochemical yield of 8% and specific activity of 21 GBq/μmol, respectively. Inhibition of the cellular uptake of these two tracers was compared by using the arabinosyl uridine analogs such as 5-iodo, 5-fluoro and 5-(E)-iodovinyl arabinosyl uridine along with 2'-fluoro-5-iodo arabinosyl uridine (FIAU). Due to potential instability of the iodo group, accumulation index of 1.6 for [(123)I]IaraU by HSV1-TK vs. control cells could virtually be achieved at 1.5 h, but dropped to 0.2 compared to 2.0 for [(18)F]FLT at 5 h. The results from competitive inhibition by these nucleosides against the accumulation of [(18)F]FLT implied that FLT exerted a mixed TK1- and TK2-dependent inhibition with HSV1-tk gene transfection because of the shifting of thymidine kinase status. Taken together, the combination of [(18)F]FLT and HSV1-TK provides a synergistic imaging potency. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... For Kids: Keep S'myelin Información en Español Brochures Free From Falls d Find Support Ask an MS ... for Change Become an MS Activist Take Action Current Advocacy Issues Advocacy Results Advocacy News d Raise ...

  4. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Programs and Events Living Well with MS Diet, Exercise & Healthy Behaviors Emotional Well-Being Spiritual Well-Being Cognitive ... Events d Living Well with MS d Diet, Exercise & Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep ...

  5. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Another Condition Aging with MS Anesthesia and Surgery Managing Your MS d Emotional Well-Being Mood Changes d Spiritual Well-Being Building Spirituality into Your Life d Cognitive Health d Work, Home & Leisure Employment Insurance & Financial Information Staying Mobile ...

  6. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Review of Society's Research Programs d Careers d Leadership Board of Directors Senior Leadership Team Founder Sylvia Lawry d Cultural Values d ... Pinterest MS Connection About the Society Vision Careers Leadership Cultural Values Financials News Press Room MS Prevalence ...

  7. Combination of PEI-Mn0.5Zn0.5Fe2O4 nanoparticles and pHsp 70-HSV-TK/GCV with magnet-induced heating for treatment of hepatoma.

    Science.gov (United States)

    Tang, Qiusha; Lu, Mudan; Chen, Daozhen; Liu, Peidang

    2015-01-01

    To explore a new combination of thermal treatment and gene therapy for hepatoma, a heat-inducible herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) gene therapy system was developed in which thermal energy generated by Mn0.5Zn0.5Fe2O4 nanoparticles (MZF-NPs) under an alternating magnetic field was used to activate gene expression. First, a recombinant eukaryotic plasmid, pHsp 70-HSV-TK, was constructed as a target gene for therapy. This recombinant plasmid was used to transfect SMMC-7721 hepatoma cells and the gene expression was evaluated. Magnet-induced heating was then applied to cells to assess the antihepatoma effects of the polyethylenimine (PEI)-MZF-NPs/pHsp 70-HSV-TK/GCV complex, in vitro and in vivo. The results showed that cells were successfully transfected with pHsp 70-HSV-TK and that expression levels of HSV-TK remained stable. Both in vitro and in vivo results indicated that the combination of gene therapy and heat treatment resulted in better therapeutic effects than heating-alone group. The rates of apoptosis and necrosis in the combined treatment group were 49.0% and 7.21%, respectively. The rate of inhibition of cell proliferation in the combined treatment group was significantly higher (87.5%) than that in the heating-alone group (65.8%; Pmagnet-induced source of heat, thereby representing a viable approach for the treatment of cancer.

  8. Combination therapy and evaluation of therapeutic effect in hepatocellular carcinoma cell using triple reporter genes; containing for NIS, HSV1-sr39tk and GFP

    Energy Technology Data Exchange (ETDEWEB)

    Lee, You La; Lee, Yong Jin; Ahn, Sohn Joo; Ahn, Byeong Cheol; Lee, Sang Woo; Yoo, Jeong Soo; Lee, Jae Tae [Kyungpook National University, Daegu (Korea, Republic of)

    2007-07-01

    To identify therapeutic effect after combine Sodium Iodine Symporter (NIS) and Mutant Herpes-simplex virus type 1 sr39tk (HSV1-sr39tk) expression in hepatocellular carcinoma cell, we transfected triple gene and investigated the properties of these gene ability in hepatocellular carcinoma cell line. After making vector with gene encoding a fusion protein comprised of HSV1-sr39tk and green florescence protein (GFP), to make triple reporter genes NIS gene was further fused to the vector using IRES vector. The vector expressing triple reporter gene was transfected to the Huh-7 cell line using liposome. Functions of hNIS and HSV1-sr39tk expression were confirmed by radio iodine uptake with and without perchlorate and [3H]-penciclovir (3-H PCV) uptake, respectively. To evaluate therapeutic effect in vitro, GCV and I-131 was treated in Huh-7/NTG cell and dual therapy performed. An animal imaging acquired using Optix and microPET in vivo. I-125 uptake was increased up to 100-fold compare to that of non-transfected cells. The transfected cell accumulated H-3 PCV up to 53 times higher at 2 hour than that of non-transfected cells. With fluorescence microscopy, green fluorescence was detected in the transfected cell. In cytotoxic studies, the cell viability of Huh-7/NTG cell was decreased to 41 % of control cell at 10ug/ml GCV concentrations. The survival rate of the Huh-7/NTG cell treated with I-131 decreased up to 16%. In I-131 and GCV dual therapy, Huh-7/NTG cell survival rate decreased up to 4%. In animal studies, Huh-7/NTG tumors showed higher uptake of 18F-FHBG and I-124 than Huh-7 tumors. GFP signal is also higher in Huh-7/NTG tumor than control. We successfully constructed a vector with delivery two therapeutic genes and one reporter gene and transfected the vector to a Huh-7 cell. The hepatocellular carcinoma cell transfected with the vector can be treated with GCV and I-131. The effect of dual gene therapy could be easily assessed by the optical reporter gene imaging.

  9. V qn TK ,

    African Journals Online (AJOL)

    Dr Obe

    RE. The conductance. G was artificially connected outside the basic built-in transistor model, as its value rises with JE current which some-time causes a convergence problem. The print-out of the program listings and computer model.

  10. V qn TK ,

    African Journals Online (AJOL)

    Dr Obe

    TEMPERATURE DEPENDENT MODELS OF SEMICONDUCTOR DEVICES FOR. COMPUTER AIDED ANALYSIS OF ELECTRONIC TRANSDUCERS. BY. TADEUSZ SIDOR. DEPARTMENT OF ELECTRICAL ENGINEERING. UNIVERSITY OF NIGERIA, NSUKKA. (Received 27th April 1981). ABSTRACT. The paper presents ...

  11. Evaluation of the Oak Ridge MS/MS explosives detector

    Energy Technology Data Exchange (ETDEWEB)

    Conrad, F.J.; Hannum, D.W.; Grant, B.C.; McLuckey, S.A.; McKown, H.S.; Glish, G.L.

    1989-01-01

    Results are presented for the evaluation of the Oak Ridge National Lab (ORNL) MS/MS as an explosives detector. This instrument is a beam instrument containing a quadrupole and a time-of-flight detector section (QTOF). The tests include a Limit of Detection for the instrument, an interferent study, a test of personnel sampling for explosives, a test involving mail sampling, and detection tests with bomb quantity explosives. 3 refs., 5 figs., 1 tab.

  12. Nuclear safety analysis for transport cask TK-6 (for WWER-440) and cover for fresh assemblies (for WWER-1000) in implementation of new fuel types at Ukrainian NPP

    International Nuclear Information System (INIS)

    Bilodid, Y.; Kovbasenko, Iu; Dudka, Olena

    2006-01-01

    According to the fresh fuel management procedure, fuel assemblies - after nuclear fuel delivery to the NPP fresh fuel unit - are vertically loaded into a cover intended for the delivery of fuel assemblies into the containment of the NPP reactor compartment. The cover is placed into an universal jack in the cooling and refueling pond, and then the fresh fuel assemblies are loaded into the reactor core. Based on the nuclear safety analysis carried out by the Russian Research Center 'Kurchatov Institute' for contemporary WWER-1000 fuel, it has become necessary to limit the number of fuel assemblies loaded into a cover below its designed capacity (12 FA instead of 18 FA as originally designed). Such a decision leads to worse economic performances in fuel transportation. The paper considers potential ways to overcome this restriction. Transport container TK-6 for spent fuel assemblies was designed quite a long time ago and, as shown in this paper, the requirement on the maximally permissible neutron multiplication factor of the loaded container for individual states to be analyzed in compliance with Ukrainian regulations is not met. First of all, this concerns the container criticality analysis in optimal neutron slow-down (container filling with water-air mixture with optimal density). The paper shows potential ways for TK-6 burnup-credit loading with the maximum number of fuel assemblies and partial container loading (Authors)

  13. MENINGKATKAN PERKEMBANGAN BAHASA INDONESIA ANAK USIA DINI MELALUI PENGGUNAAN METODE BERCERITA PADA KELOMPOK A DI TK MALAHAYATI NEUHEN TAHUN PELAJARAN 2015/2016

    Directory of Open Access Journals (Sweden)

    Rusniah Rusniah

    2017-05-01

    Full Text Available The development of language in the early childhood is the development of language that must be owned by the child as one of their basic skills, according to the stage of age and characteristics of its development. This research generally aims to improve verbal language, specifically to find out how big is the ability of spoken language through the application / use of storytelling method. This research conducted in class A TK (Kindergarten Malahayati Neuheun for Education Year 2016/2017. The subjects of the study were 22 children, 6 male students and 16 female students. The data of this research are data of students` verbal language ability, Instrument of data collection with observation sheet and data collection technique with observation. Data analysis technique used comparative descriptive method that compares cycle I and cycle II respectively in two meetings. Based on the obtained data analysis, the learning activity used II cycles, as many as 4 meetings. In the first cycle of 1st meeting the number of presentations of 36.4% was continued with the 2nd meeting of 54.5%. While on the 2nd cycle of 68% meeting followed by the 2nd meeting to 86.4%, this enhancementhas exceeded the learning mastery. The results showed that the recurring storytelling method enable to train children's concentration in improving their language skills through listening and expressing the language in group A TK Malahayati Neuheun

  14. Hydroquinone-induced malignant transformation of TK6 cells by facilitating SIRT1-mediated p53 degradation and up-regulating KRAS.

    Science.gov (United States)

    Chen, Yuting; Chen, Jiajia; Yun, Lin; Xu, Longmei; Liu, Jiaxian; Xu, Yongchun; Yang, Hui; Liang, Hairong; Tang, Huanwen

    2016-09-30

    Hydroquinone (HQ), known as one of the metabolic products of benzene, causes a number of hematologic malignancies. The study evaluated the potential mechanism of Sirtuin 1 (SIRT1) in HQ-induced TK6 cell malignant transformation. The data of our study show that short term exposure of TK6 cells to HQ led to a decrease expression of SIRT1. Knockdown of SIRT1 sensitized to the HQ-induced apoptosis in vitro and increased the expression of p53, p21 and γ-H2AX. Furthermore, chronic HQ-treated (20μM once a week for 19 weeks) caused carcinogenic transformation and was confirmed by abnormal cell proliferation, matrix metalloproteinase 9(MMP9) and subcutaneous tumor formation in nude mice. SIRT1 increased KRAS expression, and decreased H3K9 and H3K18 acetylation, inhibited p53 signaling and the level of caspase-3 in HQ-induced transformation cells. Taken together, these data suggest that SIRT1 is involved in HQ-induced malignant transformation associated with suppressing p53 signaling and activation of KRAS. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  15. miR-7-5p overexpression suppresses cell proliferation and promotes apoptosis through inhibiting the ability of DNA damage repair of PARP-1 and BRCA1 in TK6 cells exposed to hydroquinone.

    Science.gov (United States)

    Luo, Hao; Liang, Hairong; Chen, Yuting; Chen, Shaoyun; Xu, Yongchun; Xu, Longmei; Liu, Jiaxian; Zhou, Kairu; Peng, Jucheng; Guo, Guoqiang; Lai, Bei; Song, Li; Yang, Hui; Liu, Linhua; Peng, Jianming; Liu, Zhidong; Tang, Lin; Chen, Wen; Tang, Huanwen

    2018-03-01

    Hydroquinone (HQ), one of the major metabolic products of benzene, is a carcinogen, which induces apoptosis and inhibit proliferation in lymphoma cells. microRNA-7-5p (miR-7-5p), a tumor suppressor, participates in various biological processes including cell proliferation and apoptosis regulation by repressing expression of specific oncogenic target genes. To explore whether miR-7-5p is involved in HQ-induced cell proliferation and apoptosis, we assessed the effect of miR-7-5p overexpression on induction of apoptosis analyzed by FACSCalibur flow cytometer in transfection of TK6 cells with miR-7-5p mimic (TK6- miR-7-5p). We observed an increased apoptosis by 25.43% and decreased proliferation by 28.30% in TK6-miR-7-5p cells compared to those negative control cells (TK6-shNC) in response to HQ treatment. Furthermore, HQ might active the apoptotic pathway via partly downregulation the expression of BRCA1 and PARP-1, followed by p53 activation, in TK6-miR-7-5p cells. In contrast, attenuated p53 and BRCA1 expression was observed in shPARP-1 cells than in NC cells after HQ treatment. Therefore, we conclude that HQ may activate apoptotic signals via inhibiting the tumor suppressive effects of miR-7-5p, which may be mediated partly by upregulating the expression of PARP-1 and BRCA1 in control cells. The increase of miR-7-5p expression further intensified downregulation of PARP-1 and BRCA1 in TK6-miR-7-5p cells, resulting in an increase of apoptosis and proliferation inhibited. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Employer Matching Gifts d Gifts of Stock or Securities d Giving Circles Golden Circle Circle of Distinction ... and problems with cognition (learning and memory or information processing). People with progressive forms of MS are ...

  17. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Researchers Get Involved Fundraising Events Volunteer Advocate for Change Raise Awareness Join the Community Stay Informed Corporate ... Managing Your MS d Emotional Well-Being Mood Changes d Spiritual Well-Being Building Spirituality into Your ...

  18. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Senior Leadership Team Founder Sylvia Lawry d Cultural Values d Financials Annual Reports Sources of Support d ... Connection About the Society Vision Careers Leadership Cultural Values Financials News Press Room MS Prevalence Charitable Ratings ...

  19. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Treating MS d Comprehensive Care Developing a Healthcare Team Make the Most of Your Doctor Visits Advance ... Stay Informed d Corporate Support Corporate Partners National Teams Partnership Opportunities d Personal Stories Ambassadors & Familiar Faces ...

  20. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... d Other Conditions to Rule Out Lyme Disease Lupus Neuromyelitis Optica Acute Disseminated Encephalomyelitis (ADEM) d For ... with MS d Diet, Exercise & Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health ...

  1. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Healthy Behaviors Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and ... Its Tracks Restoring What's Been Lost Ending the Disease Forever How We Fund Research FAQs about Society ...

  2. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Behaviors Emotional Well-Being Spiritual Well-Being Cognitive Health Work, Home & Leisure Relationships Research Participate in Research ... Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another Condition Aging ...

  3. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... also called plaques or scars — on magnetic resonance imaging (MRI) scans, and these lesions contain more inflammatory cells. People with primary progressive MS (PPMS) tend to ...

  4. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Relationships Research Participate in Research Studies Research News & Progress Research We Fund ... Hispanics & Latino/as d Multiple Sclerosis FAQs d Types of MS Clinically Isolated Syndrome ( ...

  5. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Relapsing-remitting MS (RRMS) Diagnosing RRMS Treating RRMS Research in RRMS Share Smaller Text Larger Text Print In this article Overview RRMS – the most common disease course – is ...

  6. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... the disease during the periods of remission. At different points in time, RRMS can be further characterized ... to characterize the course of your disease at different points in time helps you and your MS ...

  7. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... membranes surrounding nerve fibers in the central nervous system (CNS)), as well as the nerve fibers themselves. ... and problems with cognition (learning and memory or information processing). People with progressive forms of MS are ...

  8. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Online d Give in Honor or Memory d Workplace Giving d Employer Matching Gifts d Gifts of ... Brochure Working with MS (.pdf) Download Brochure Taming Stress (.pdf) Download Brochure Multiple Sclerosis and Your Emotions (. ...

  9. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... discuss your treatment options and expected outcomes. For example: If you have RRMS that is active and worsening, you and your MS care provider will likely want to consider a more aggressive treatment approach than if there ...

  10. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Events Living Well with MS Diet, Exercise & Healthy Behaviors Emotional Well-Being Spiritual Well-Being Cognitive Health Work, Home & Leisure Relationships Research Participate in Research Studies Research News & Progress Research We Fund For Researchers ...

  11. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Find Support Advanced Care Needs Resources for Specific Populations Find Programs & Services in Your Area Calendar of ... Services: Questions to Ask d Resources for Specific Populations Pediatric MS Support Veterans with Multiple Sclerosis d ...

  12. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Disasters d Relationships Disclosure Family Matters Parenting Intimacy Self Advocacy in the Family Preventing Abuse Testing for ... Our MS Navigators help identify solutions and provide access to the resources you are looking for. Call ...

  13. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... FAQs about Society Research Past Research Efforts Driving Solutions d For Researchers d Get Involved d Volunteer ... We Are Here Our MS Navigators help identify solutions and provide access to the resources you are ...

  14. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Diet & Nutrition Exercise Heat & Temperature Sensitivity Sleep Vaccinations Women's Health Unhealthy Habits Managing MS and Another Condition ... lesions, which contain fewer inflammatory cells. In RRMS, women are affected two to three times as often ...

  15. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Directors Senior Leadership Team Founder Sylvia Lawry d Cultural Values d Financials Annual Reports Sources of Support d News d Press Room Events at a Glance MS the Disease Public Service ...

  16. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Research Diet Vitamin D How and Why Do Scientists Share Results d Research We Fund Stopping MS ... Share Smaller Text Larger Text Print In this article Overview RRMS – the most common disease course – is ...

  17. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Services in Your Area Calendar of Programs and Events Living Well with MS Diet, Exercise & Healthy Behaviors ... Research We Fund For Researchers Get Involved Fundraising Events Volunteer Advocate for Change Raise Awareness Join the ...

  18. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Fellows d Careers in MS Clinical Care Research Have you met? d Our Healthcare Voice Home What ... the damage is so variable, no two people have exactly the same symptoms. Why are modifiers used ...

  19. MRI atlas of MS lesions

    Energy Technology Data Exchange (ETDEWEB)

    Sahraian, Mohammad Ali [Tehran Univ. of Medical Sciences Sina Hospital (Iran). Dept. of Neurology; Radue, Ernst-Wilhelm [Univ. Hospital Basel (Switzerland). Dept. of Neuroradiology

    2008-07-01

    MRI has become the main paraclinical test in the diagnosis and management of multiple sclerosis. We have demonstrated more than 400 pictures of different typical and atypical MS lesions in this atlas. Each image has a teaching point. New diagnostic criteria and differential diagnosis are discussed and the book is supported by a teaching DVD where the reader can see MS lesions in different slices and sequences. (orig.)

  20. Gene expression profiles and genetic damage in benzo(a)pyrene diol epoxide-exposed TK6 cells

    Energy Technology Data Exchange (ETDEWEB)

    Akerman, G.S.; Rosenzweig, B.A.; Domon, O.E.; McGarrity, L.J.; Blankenship, L.R.; Tsai, C.A.; Culp, S.J.; MacGregor, J.T.; Sistare, F.D.; Chen, J.J.; Morris, S.M

    2004-05-18

    Microarray analysis is a powerful tool to identify the biological effects of drugs or chemicals on cellular gene expression. In this study, we compare the relationships between traditional measures of genetic toxicology and mutagen-induced alterations in gene expression profiles. TK6 cells were incubated with 0.01, 0.1, or 1.0 {mu}M {+-}anti-benzo(a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide (BPDE) for 4 h and then cultured for an additional 20 h. Aliquots of the exposed cells were removed at 4 and 24 h in order to quantify DNA adduct levels by {sup 32}P post-labeling and measure cell viability by cloning efficiency and flow cytometry. Gene expression profiles were developed by extracting total RNA from the control and exposed cells at 4 and 24 h, labeling with Cy3 or Cy5 and hybridizing to a human 350 gene array. Mutant frequencies in the Thymidine Kinase and Hypoxanthine Phosphoribosyl Transferase genes were also determined. The 10{alpha}-(deoxyguanosin-N{sup 2}-yl)-7{alpha},8{beta},9{beta}-trihydroxy-7,8,9,10-= tetrahydrobenzo(a) pyrene (dG-N{sup 2}-BPDE) adduct increased as a function of dose and was the only adduct identified. A dose-related decrease in cell viability was evident at 24 h, but not at 4 h. Cell death occurred by apoptosis. At 4 h, analysis of the gene expression profiles revealed that Glutathione Peroxidase and Gadd45 were consistently upregulated (greater than 1.5-fold and significantly (P<0.001) greater than the control in two experiments) in response to 1.0 {mu}M BPDE exposure. Fifteen genes were consistently down-regulated (less than 0.67-fold and significantly (P<0.001) lower than the control in two experiments) at 4 h in cultures exposed to 1.0 {mu}M BPDE. Genes with altered expression at 4 h included genes important in the progression of the cell-cycle and those that inhibit apoptosis. At 24 h post-exposure, 16 genes, involved in cell-cycle control, detoxification, and apoptosis were consistently upregulated; 10 genes were repressed in

  1. An IMS-IMS Analogue of MS-MS

    Energy Technology Data Exchange (ETDEWEB)

    Koeniger, Stormy L.; Merenbloom, Samuel I.; Valentine, Stephen J.; Jarrold, Martin F.; Udseth, Harold R.; Smith, Richard D.; Clemmer, David E.

    2006-06-15

    The development of a new ion mobility/mass spectrometry instrument that incorporates a multifield drift tube/ion funnel design is described. In this instrument, individual components from a mixture of ions can be resolved and selected on the basis of mobility differences prior to collisional activation inside the drift tube. The fragment ions that are produced can be dispersed again in a second ion mobility spectrometry (IMS) region prior to additional collisional activation and MS analysis. The result is an IMS-IMS analogue of MS-MS. Here, we describe the preliminary instrumental design and experimental approach. We illustrate the approach by examining the highly characterized bradykinin and ubiquitin systems. Mobility-resolved fragment ions of bradykinin show that b-type ions are readily discernible fragments, because they exist as two easily resolvable structural types. Current limitations and future directions are briefly discussed.

  2. RETRACTED : Upaya Meningkatkan Kemampuan Fisik Motorik Kasar Anak melalui Kegiatan Melambungkan dan Menangkap Dengan Berbagai Media Anak Usia Dini di TK Al- Fajar Pekanbaru

    Directory of Open Access Journals (Sweden)

    Melvi Lesmana Alim

    2016-06-01

    Full Text Available Following a rigorous, carefully concerns and considered review of the article published in Jurnal Obsesi to article entitled “Upaya Meningkatkan Kemampuan Fisik Motorik Kasar Anak melalui Kegiatan Melambungkan dan Menangkap dengan Berbagai Media Anak Usia Dini di TK Al- Fajar Pekanbaru” Vol 2, No 2, pp. 79-89, December 2016, DOI: https://doi.org/10.2155/obsesi.v2i1.120   This paper has been found to be in violation of the International Journal of Advances in Intelligent Informatics Publication principles and has been retracted.   The article contained redundant material, the editor investigated and found that the paper published in Jurnal Obsesi, Vol. 1, No. 2, pp. 1-11, december 2015.   The document and its content has been removed from International Journal of Advances in Intelligent Informatics, and reasonable effort should be made to remove all references to this article.

  3. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Series Momentum Magazine Educational Videos Knowledge Is Power Live Fully, Live Well Everyday Matters Resilience: Addressing the Challenges of ... and Surgery Managing Your MS d Emotional Well-Being Mood Changes d Spiritual Well-Being Building Spirituality ...

  4. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... bowel and bladder problems, and problems with cognition (learning and memory or information processing). People with progressive ... Colophon Stay Informed Join Us Facebook Twitter LinkedIn YouTube Pinterest MS Connection About the Society Vision Careers ...

  5. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... the Challenges of MS You CAN! Webcasts DVDs Books For Kids: Keep S'myelin Información en Español Brochures ... RRMS Treating RRMS Research in RRMS Share Smaller Text Larger Text Print In this article Overview RRMS – ...

  6. Relapsing-Remitting MS (RRMS)

    Medline Plus

    Full Text Available ... Multiple Sclerosis and Your Emotions (.pdf) Download Brochure Disease Modifying Therapies Overview (.pdf) Download Document Pediatric MS Learn More Transverse Myelitis Learn More Neuromyelitis Optica (NMO) Learn More HTLV-I ... Learn More Acute Disseminated Encephalomyelitis (ADEM) Learn More ...

  7. Method-MS. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Skipperud, L.; Popic, J.M. (Norwegian Univ. of Life Science (UMB), Isotope Lab. (Norway)); Roos, P. (Technical Univ. of Denmark, Risoe National Lab. for Sustainable Energy, Roskilde (Denmark)); Salminen, S. (Univ. of Helsinki (UH) (Finland)); Nygren, U. (Swedish Defence Research Agency (FOI) (Sweden)); Sigmarsson, O.; Palsson, S.E. (Univ. of Iceland/Icelandic Radiation Protection Institute (Iceland))

    2011-05-15

    Radiometric determination methods, such as alpha spectrometry require long counting times when low activities are to be determined. Mass spectrometric techniques as Inductively Coupled Plasma Mass Spectrometry (ICP-MS), Thermal Ionisation Mass Spectrometry (TIMS) and Accelerator Mass Spectrometry (AMS) have shown several advantages compared to traditional methods when measuring long-lived radionuclides. Mass spectrometric methods for determination of very low concentrations of elemental isotopes, and thereby isotopic ratios, have been developed using a variety of ion sources. Although primarily applied to the determination of the lighter stable element isotopes and radioactive isotopes in geological studies, the techniques can equally well be applied to the measurement of activity concentrations of long-lived low-level radionuclides in various samples using 'isotope dilution' methods such as those applied in inductively coupled plasma mass spectrometry (ICP-MS). Due to the low specific activity of long-lived radionuclides, many of these are more conveniently detected using mass spectrometric techniques. Mass spectrometry also enables the individual determination of Pu-239 and Pu-240, which cannot be obtained by alpha spectrometry. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) are rapidly growing techniques for the ultra-trace analytical determination of stable and long-lived isotopes and have a wide potential within environmental science, including ecosystem tracers and radio ecological studies. Such instrumentation, of course needs good radiochemical separation, to give best performance. The objectives of the project is to identify current needs and problems within low-level determination of long-lived radioisotopes by ICP-MS, to perform intercalibration and development and improvement of ICP-MS methods for the measurement of radionuclides and isotope ratios and to develop new methods based on modified separation chemistry applied to new

  8. Determination of paraquat in vegetables using HPLC-MS-MS.

    Science.gov (United States)

    Zou, Tingting; He, Pingli; Cao, Jingjing; Li, Zhen

    2015-02-01

    A simple, sensitive, reliable and economical method was developed for the determination of paraquat (a widely used herbicide) in four edible vegetables (cabbage, lettuce, spinach and Chinese cabbage) using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS). The samples were extracted with water under sonication and cleaned up by weak cation exchange solid-phase extraction. Chromatographic separation of paraquat was achieved on a hydrophilic interaction liquid chromatography column (2.1 × 100 mm, 3 µm) with a gradient program using 10 mM ammonium acetate in 0.1% formic acid and acetonitrile as mobile phase. The low salt concentration used in the eluting buffer ensured extended LC-MS analysis of paraquat in different matrices without the necessity of frequent source cleaning. The validity of the developed method was evaluated by spiking paraquat in four edible vegetables at 50 and 500 ng g(-1). Recovery ranged from 43.6 to 73.5%. The limit of detection is 0.94 ng g(-1). With the developed method, the kinetic of paraquat entering plant tissue was also evaluated. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  9. Analytical Method Details (MS): SE24_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE24_MS1 LC-FTICR-MS Analysis (Positive) LC, Agilent 1200 series (Agilent); MS, Sol...sample (1 uL) was analyzed using LC-FTICR-MS (LC, Agilent 1200 series; MS, Bruker Daltonics solarix 7.0 T). ...olarity, positive; calibrant, low concentration TuningMix (Agilent); lock mass target, diethylhexylphthalate

  10. LC-MS based Metabolomics

    DEFF Research Database (Denmark)

    Magdenoska, Olivera

    metabolites with liquid chromatography mass spectrometry (LC-MS) as the most commonly used. The primary goal of this Ph.D. study was to develop an LC-MS method together with sample preparation for analysis of intracellular metabolites such as nucleotides, sugar phosphates, organic acids, coenzymes etc...... during this Ph.D. study were successfully applied for targeted and multitargeted analysis of different classes of intracellular metabolites such as nucleotides, sugar phosphates, coeznymes and organic acids. In addition sample preparation methods were established for different microorganisms capable...... recovery of the metabolites after the extraction. Quenching and extraction procedures for bacteria, yeast, mammalian cells and filamentous fungi were tested. Cold MeOH as a quenching method combined with boiling EtOH or MeOH/chloroform as extraction method showed to work well for Saccharomyces cerevisiae...

  11. Analysis of phytochemical variations in dioecious Tinospora cordifolia stems using HPLC/QTOF MS/MS and UPLC/QqQLIT -MS/MS.

    Science.gov (United States)

    Bajpai, Vikas; Singh, Awantika; Chandra, Preeti; Negi, M P S; Kumar, Nikhil; Kumar, Brijesh

    2016-01-01

    The stem of dioecious Tinospora cordifolia (Menispermaceae) is a commonly used traditional Ayurvedic medicine in India having several therapeutic properties. To develop and validate LC-MS methods for the identification and simultaneous quantitation of various secondary metabolites and to study metabolomic variations in the stem of male and female plants. Ethanolic extract of stems were analysed by HPLC/ESI-QTOF-MS/MS for rapid screening of bioactive phytochemicals. High resolution MS and MS/MS in positive ESI mode were used for structural investigation of secondary metabolites. An UPLC/ESI-QqQ(LIT) -MS/MS method in MRM mode was developed and validated for the simultaneous quantitation of five bioactive alkaloids. Identification and characterisation of 36 metabolites including alkaloids, sesquiterpenes and phytoecdysteroids were performed using LC-MS and MS/MS techniques. The bioactive alkaloids such as jatrorrhizine, magnoflorine, isocorydine, palmatine and tetrahydropalmatine were successfully quantified in male and female plants. The mean abundances of magnoflorine jatrorrhizine, and oblongine were significantly (P Phytochemicals in the stem of male and female Tinospora cordifolia showed significant qualitative and quantitative variations. LC-MS and MS/MS methods can be used to differentiate between male and female plants based on their chemical profiles and quantities of the marker bioactive alkaloids. This chemical composition difference was also evident during vegetative stage when there were no male and female flowers. Copyright © 2015 John Wiley & Sons, Ltd.

  12. PTR-MS in enology

    Science.gov (United States)

    Spitaler, Renate; Araghipour, Nooshin; Mikoviny, Tomas; Wisthaler, Armin; Via, Josef Dalla; Märk, Tilmann D.

    2007-10-01

    The present communication deals with the improvement of proton transfer reaction mass spectrometry (PTR-MS) wine headspace analyses. In contrast to previous PTR-MS investigations of wine, where wine headspace was ionized by protonated ethanol clusters, the headspace was diluted by a factor of 1:40 with N2 and ionized by H3O+ ions. This method is better suited for routine applications than the previously reported method since it is simpler, faster, and the mass spectra obtained are less complex. A test wine was mixed with ethanol and with water to yield ethanol contents ranging from 10 to 15% (v/v) and these mixtures were analyzed to assess whether any quantitative differences in the composition of volatiles were detectable. The data showed no impact of the ethanol content on the wine headspace composition. The new method was applied to eight different wine samples produced from two different grape varieties: Pinot Noir and Cabernet Sauvignon. Each variety was grown in two different locations in South Tyrol (Northern Italy) and harvested at two different dates. Quantitative (but not qualitative) differences in PTR-MS spectra between the two wine varieties were observed. Using principal component analysis of selected m/z signals differentiation between Pinot Noir and Cabernet Sauvignon samples was achievable.

  13. Effects of structural differences between radioiodine-labeled 1-(2'-fluoro-2'-deoxy-D-arabinofuranosyl)-5-iodouracil (FIAU) and 1-(2'-fluoro-2'-deoxy-D-ribofuranosyl)-5-iodouracil (FIRU) on HSV1-TK reporter gene imaging

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Eun Jung; Hong, Su Hee [Radiopharmaceutical Research Team, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of); Choi, Tae Hyun [Radiopharmaceutical Research Team, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of)], E-mail: thchoi@kirams.re.kr; Lee, Eun Ah [Radiopharmaceutical Research Team, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of); Kim, Kyeong Min [Nuclear Medicine Research Team, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of); Lee, Kyo Chul; An, Gwang Il [Radiopharmaceutical Research Team, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of); El-Gamal, Mohammed I. [Biomaterials Research Center, Korea Institute of Science and Technology, PO Box 131, Cheongryang, Seoul 130-650 (Korea, Republic of); Department of Biomolecular Science, University of Science and Technology, 113 Gwahangno, Yuseong-gu, Daejeon 305-333 (Korea, Republic of); Cheon, Gi Jeong [Radiopharmaceutical Research Team, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of); Department of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of); Choi, Chang Woon; Lim, Sang Moo [Department of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706 (Korea, Republic of)

    2010-06-15

    The goal of this investigation was to evaluate the effects of structural differences between FIAU and FIRU on their ability to serve as a potential tracer for reporter gene imaging. To examine the characteristics of different configurations of FIAU and FIRU, a series of evaluations were done on HSV1-TK gene-expressing cells and on mice with HSV1-TK gene-expressing tumor. The results showed that, as an imaging agent for HSV1-TK-expressing cells, radiolabeled FIAU was more efficient for in vivo imaging than FIRU.

  14. Selenium speciation analysis of Misgurnus anguillicaudatus selenoprotein by HPLC-ICP-MS and HPLC-ESI-MS/MS

    Science.gov (United States)

    Analytical methods for selenium (Se) speciation were developed using high performance liquid chromatography (HPLC) coupled to either inductively coupled plasma mass spectrometry (ICP-MS) or electrospray ionization tandem mass spectrometry (ESI-MS/MS). Separations of selenomethionine (Se-Met) and sel...

  15. Molecular analysis of intact preen waxes of Calidris Canutus (Aves: Scolopacidae) by GC/MS and GC/MS/MS

    NARCIS (Netherlands)

    Sinninghe Damsté, J.S.; Dekker, M.H.A.; Piersma, T.

    2000-01-01

    The intact preen wax esters of the red knot Calidris canutus were studied with gas chromatography/mass spectrometry (GC/MS) and GC/MS/MS. In this latter technique, transitions from the molecular ion to fragment ions representing the fatty acid moiety of the wax esters were measured, providing

  16. Familial Incidence of Multiple Sclerosis (MS)

    OpenAIRE

    J Gordon Millichap

    1988-01-01

    Age-adjusted familial rates for MS were determined in children and siblings of patients studied at the MS Clinic and Dept of Genetics, Health Sciences Centre Hospital, Univ British Columbia, Vancouver BC, Canada.

  17. EGFR-TK inhibition before radiotherapy reduces tumour volume but does not improve local control: Differential response of cancer stem cells and nontumourigenic cells?

    International Nuclear Information System (INIS)

    Krause, Mechthild; Prager, Jenny; Zhou Xuanjing; Yaromina, Ala; Doerfler, Annegret; Eicheler, Wolfgang; Baumann, Michael

    2007-01-01

    Background and purpose: Waiting times before radiotherapy may reduce tumour control probability due to proliferation of tumour cells. The aim of the experiment was to test whether the growth inhibiting effect of epidermal growth factor receptor (EGFR)-inhibitors after surgery or tumour transplantation results in a lower tumour mass at time of irradiation and can thereby improve local tumour control. Materials and methods: The EGFR-tyrosine kinase inhibitor BIBX1382BS was applied over 14 days starting from microscopically non-in-sano-resection of FaDu tumours or from tumour transplantation, followed by irradiation (5f/5d). Endpoint was local tumour control. In addition, vital tumour areas, pimonidazole hypoxic fraction, BrdU labelling index, and colony forming ability in vitro were tested in control tumours and after BIBX1382BS treatment (starting from transplantation). Results: The tumour volume at start of irradiation was significantly lower in the BIBX1382BS treated tumours as compared to the control groups by factors of 11 (post-surgery setting) and 2.7 (transplantation setting). However, the reduced volume did not translate into improved local control after irradiation. The TCD 50 values after surgery were 25.4 Gy [95% CI 18; 33 Gy] in the control group and 30.5 Gy [24; 37] in the BIBX1382BS group (p = 0.25). Treatment after transplantation resulted in TCD 50 values of 41.1 Gy [35; 47] in the control group and 41.1 Gy [33; 49] in the BIBX1382BS group (p = 1). While the proportion of S-phase cells decreased after BIBX1382BS treatment, no differences were observed between the pimonidazole hypoxic fractions and in vitro colony forming ability. Conclusions: EGFR-TK inhibition with BIBX1382BS over 14 days between macroscopically complete tumour resection or tumour transplantation and start of radiotherapy significantly reduced tumour volume but did not improve local tumour control. One possible explanation is that the EGFR-TK inhibitor has a higher activity in

  18. Physical Exercise and MS Recommendations

    DEFF Research Database (Denmark)

    Dalgas, U; Ingemann-Hansen, T; Stenager, E

    2009-01-01

    The use of physical exercise programmes in the rehabilitation of patients with multiple sclerosis (MS) has been a controversial issue for many years. During the last decade, however, evidence from a number of studies has suggested that exercise is a safe and efficient way to induce improvements...... in a number of physiological functions, which ultimately can lead to functional improvements that have a positive effect on a patients daily life. The purpose of this review is, based on the existing research, to provide clinicians with some easily administrable recommendations for the application of exercise...

  19. Physical Exercise and MS Recommendations

    DEFF Research Database (Denmark)

    Dalgas, U; Ingemann-Hansen, T; Stenager, E

    2009-01-01

    in a number of physiological functions, which ultimately can lead to functional improvements that have a positive effect on a patients daily life. The purpose of this review is, based on the existing research, to provide clinicians with some easily administrable recommendations for the application of exercise......The use of physical exercise programmes in the rehabilitation of patients with multiple sclerosis (MS) has been a controversial issue for many years. During the last decade, however, evidence from a number of studies has suggested that exercise is a safe and efficient way to induce improvements...

  20. Combination of PEI-Mn0.5Zn0.5Fe2O4 nanoparticles and pHsp 70-HSV-TK/GCV with magnet-induced heating for treatment of hepatoma

    OpenAIRE

    Tang QS; Lu MD; Chen DZ; Liu PD

    2015-01-01

    Qiusha Tang,1 Mudan Lu,2 Daozhen Chen,2 Peidang Liu1 1School of Medicine, Southeast University, Nanjing, 2Genetic Laboratory, Wuxi Hospital for Maternal and Child Health Care, the Affiliated Hospital of Nanjing Medical University, Wuxi, People’s Republic of China Background: To explore a new combination of thermal treatment and gene therapy for hepatoma, a heat-inducible herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) gene therapy system was developed in wh...

  1. The Epstein-Barr virus (EBV)-encoded protein kinase, EBV-PK, but not the thymidine kinase (EBV-TK), is required for ganciclovir and acyclovir inhibition of lytic viral production.

    Science.gov (United States)

    Meng, Qiao; Hagemeier, Stacy R; Fingeroth, Joyce D; Gershburg, Edward; Pagano, Joseph S; Kenney, Shannon C

    2010-05-01

    Ganciclovir (GCV) and acyclovir (ACV) are guanine nucleoside analogues that inhibit lytic herpesvirus replication. GCV and ACV must be monophosphorylated by virally encoded enzymes to be converted into nucleotides and incorporated into viral DNA. However, whether GCV and/or ACV phosphorylation in Epstein-Barr virus (EBV)-infected cells is mediated primarily by the EBV-encoded protein kinase (EBV-PK), the EBV-encoded thymidine kinase (EBV-TK), or both is controversial. To examine this question, we constructed EBV mutants containing stop codons in either the EBV-PK or EBV-TK open reading frame and selected for stable 293T clones latently infected with wild-type EBV or each of the mutant viruses. Cells were induced to the lytic form of viral replication with a BZLF1 expression vector in the presence and absence of various doses of GCV and ACV, and infectious viral titers were determined by a green Raji cell assay. As expected, virus production in wild-type EBV-infected 293T cells was inhibited by both GCV (50% inhibitory concentration [IC(50)] = 1.5 microM) and ACV (IC(50) = 4.1 microM). However, the EBV-PK mutant (which replicates as well as the wild-type (WT) virus in 293T cells) was resistant to both GCV (IC(50) = 19.6 microM) and ACV (IC(50) = 36.4 microM). Expression of the EBV-PK protein in trans restored GCV and ACV sensitivity in cells infected with the PK mutant virus. In contrast, in 293T cells infected with the TK mutant virus, viral replication remained sensitive to both GCV (IC(50) = 1.2 microM) and ACV (IC(50) = 2.8 microM), although susceptibility to the thymine nucleoside analogue, bromodeoxyuridine, was reduced. Thus, EBV-PK but not EBV-TK mediates ACV and GCV susceptibilities.

  2. Aktualisasi Pendidikan Al-Qur’an melalui Kegiatan Habit-Forming pada Anak Usia Dini di TK Al-Kautsar Durisawo Ponorogo

    Directory of Open Access Journals (Sweden)

    RISA ALFIYAH ULFA

    2017-12-01

    Full Text Available This study is based on the phenomenon that education of the Qur'an in the Muslim community is lower. And its actualization into formal and non formal education is needed, one of them through habit-forming activity. This study aims to determine the actualization of education of the Qur'an through habit-forming activities and its implications for reading al-Qur'an ability in TK al-Kautsar Durisawo Ponorogo. This research is qualitative with psychologycal approach using behaviorism theory of learning . and data colecting proses using interviews, observation, documentation studies, and triangulation. The result of this research is known that the actualization of education of al-Qur'an through habit-forming activity in kindergarten Al-Kautsar Durisawo Ponorogo, such as in the meaning of recitations more emphasize the concept of reading in tahqiq. In the meaning of tadabbur, there is the provision of Islamic stories. In the meaning of tahfidz, it is limited to memorizing short letters contained in juz 'Amma. In the meaning of practice, habituation utter and answer greetings. In the meaning of nurturing, applied through the practice of praying fardhu congregation who always memorized the prayer readings, especially the letter al-Fatihah. The implication, learners confident, firm, meticulous in reading hijaiyah letters, strengthen memorization of learners, especially letter al-Fatihah. The habit of praying sunnah dhuha in congregation to form an independent attitude in applying the al-Fatihah letters

  3. Reading Culture of Pre-service Turkish Language Teachers: The Case of Muğla Sıtkı Koçman University

    Directory of Open Access Journals (Sweden)

    Hatice FIRAT

    2017-04-01

    Full Text Available This study aims to reveal the reading culture and determine the characteristics and dispositions of pre-service Turkish language teachers as readers. The study was conducted with 160 pre-service Turkish language teachers studying in the Department of Turkish Language Teaching at Muğla Sıtkı Koçman University’s Faculty of Education in Turkey. The study took place during the 2015-2016 academic year and all participants were volunteers. The data were obtained through questionnaires, and data analysis was conducted by using frequency and cross tabulation analysis. Pre-service teachers’ responses were analyzed using the content analysis method. According to the results of the study; it was identified that pre-service Turkish language teachers prefer “reading books” in order to make good use of their leisure time and also their teachers and friends have some influence on the development of their "reading habit" and "reading culture". Additionally, the pre-service teachers proposed "doing research as a course requirement" as another reason for their reading. Nonetheless, they remarked that they prefer “literary works” among the book types and especially “novels”. In addition, the pre-service teachers stated that they often prefer "borrowing from the library" to buying books they would like to read. It was also determined that they also read an average of 11-20 books in a year and therefore they might be classified as "medium level readers".

  4. A high-resolution whole genome radiation hybrid map of human chromosome 17q22-q25.3 across the genes for GH and TK

    Energy Technology Data Exchange (ETDEWEB)

    Foster, J.W.; Schafer, A.J.; Critcher, R. [Univ. of Cambridge (United Kingdom)] [and others

    1996-04-15

    We have constructed a whole genome radiation hybrid (WG-RH) map across a region of human chromosome 17q, from growth hormone (GH) to thymidine kinase (TK). A panel of 128 WG-RH hybrid cell lines generated by X-irradiation and fusion has been tested for the retention of 39 sequence-tagged site (STS) markers by the polymerase chain reaction. This genome mapping technique has allowed the integration of existing VNTR and microsatellite markers with additional new markers and existing STS markers previously mapped to this region by other means. The WG-RH map includes eight expressed sequence tag (EST) and three anonymous markers developed for this study, together with 23 anonymous microsatellites and five existing ESTs. Analysis of these data resulted in a high-density comprehensive map across this region of the genome. A subset of these markers has been used to produce a framework map consisting of 20 loci ordered with odds greater than 1000:1. The markers are of sufficient density to build a YAC contig across this region based on marker content. We have developed sequence tags for both ends of a 2.1-Mb YAC and mapped these using the WG-RH panel, allowing a direct comparison of cRay{sub 6000} to physical distance. 31 refs., 3 figs., 2 tabs.

  5. Acute Respiratory Distress due to Thymoma in a Patient Treated with TK Inhibitor: A Case Report and Review of the Current Treatment Options

    Directory of Open Access Journals (Sweden)

    P. Zarogoulidis

    2011-03-01

    Full Text Available Thymic malignancies are rare intrathoracic tumors that may be aggressive and difficult to treat in advanced stage. Surgery is the cornerstone of the management of thymomas: it is significant for the definite histopathological diagnosis and staging, and in most cases, it constitutes the first step of the treatment strategy. For patients with primary unresectable thymomas, the multimodal treatment schedule nowadays includes neoadjuvant chemotherapy, extensive surgery, adjuvant radiotherapy, and in some cases, adjuvant chemotherapy. A patient with a history of stage III COPD and an undiagnosed thoracic mass was admitted to the intensive care unit with acute respiratory distress. A radiologic evaluation by CT scan revealed a mass of 13 cm in diameter at the mediastinum. Fine needle aspiration was performed and revealed a thymoma. Due to poor performance status, the patient was not able to undergo surgery. He refused to be treated with neither chemotherapy nor radiotherapy, but due to EGFR overexpression, treatment with TK inhibitor was suggested. Fine needle aspiration biopsy is commonly used to identify metastasis to the mediastinum. However, it is less often employed as a primary diagnostic tool for tumors, particularly thymic neoplasms. The use of targeted therapies for the treatment of thymic malignancies has been described in the literature. Over the past years, significant efforts have been made to dissect the molecular pathways involved in the carcinogenesis of these tumors. Insights have been obtained following anecdotal clinical responses to targeted therapies, and large-scale genomic analyses have been conducted.

  6. Evaluation of phenolphthalein, diazepam and quinacrine dihydrochloride in the in vitro mammalian cell micronucleus test in Chinese hamster ovary (CHO) and TK6 cells.

    Science.gov (United States)

    Schuler, Maik; Gudi, Ramadevi; Cheung, Jennifer; Kumar, Shyam; Dickinson, Donna; Engel, Maria; Szkudlinska, Anna; Colman, Megan; Maduka, Nkechi; Sherman, Jocelyn; Thiffeault, Catherine

    2010-10-29

    The in vitro micronucleus assay has been extensively used as an in vitro screening tool to identify test articles that might have aneugenic or clastogenic potential. Currently, the Organization for Economic Co-operation and Development (OECD) is working towards a final version of the guideline for the conduct of the in vitro mammalian cell micronucleus Test (MNvit). A few questions regarding appropriate cytotoxicity measurements and cytotoxicity limits to use remain to be answered. In order to resolve the remaining questions, we compared the induction of micronuclei at the top dose (50-60% cytotoxicity) determined by either Relative Cell Counts (RCC), Relative Increase in Cell Counts (RICC), Relative Population Doublings (RPD), or Cytokinesis-Blocked Proliferating Index (CBPI) using weak and strong inducers of micronuclei in both the presence and absence of cytochalasin B (CYB) in Chinese hamster ovary (CHO) and human lymphoblastoid TK6 cells. In order to assess extensive dose-response relationships, we selected expected weak (diazepam, phenolphthalein, quinacrine dihydrochloride dihydrate) and strong (cytosine arabinoside, mitomycin C, vinblastine sulphate) inducers of micronuclei with a variety of different mechanisms of action for testing. The results clearly demonstrated that all six compounds produced positive responses using either cytotoxicity measurement. The outcome from these studies further supports the cytotoxicity measurements and cytotoxicity limits proposed in the draft OECD guideline. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. A comprehensive full factorial LC-MS/MS proteomics benchmark data set.

    NARCIS (Netherlands)

    Wessels, H.J.C.T.; Bloemberg, T.G.; Dael, M.F.P. van; Wehrens, R.; Buydens, L.M.; Heuvel, L.P.W.J. van den; Gloerich, J.

    2012-01-01

    An important prerequisite for the development and benchmarking of novel analysis methods is a well-designed comprehensive LC-MS/MS data set. Here, we present our data set consisting of 59 LC-MS/MS analyses of 50 protein samples extracted individually from Escherichia coli K12 and spiked with

  8. Direct sampling tandem mass spectrometry (MS/MS) and multiway calibration for isomer quantitation

    NARCIS (Netherlands)

    Zampronio, Cleidiane G.; Gurden, Stephen P.; Moraes, Luis Alberto B.; Eberlin, Marcos N.; Smilde, Age K.; Poppi, Ronei J.

    2002-01-01

    Direct sampling tandem mass spectrometry (MS/MS) was used for the quantitation of mixtures of the isomers 2-, 3- and 4-ethyl pyridine. The similarity between the analytes and the second-order nature of MS/MS data require the use of multivariate calibration techniques capable of handling multiway

  9. Noninvasive theranostic imaging of HSV1-sr39TK-NTR/GCV-CB1954 dual-prodrug therapy in metastatic lung lesions of MDA-MB-231 triple negative breast cancer in mice.

    Science.gov (United States)

    Sekar, Thillai V; Foygel, Kira; Ilovich, Ohad; Paulmurugan, Ramasamy

    2014-01-01

    Metastatic breast cancer is an obdurate cancer type that is not amenable to chemotherapy regimens currently used in clinic. There is a desperate need for alternative therapies to treat this resistant cancer type. Gene-Directed Enzyme Prodrug Therapy (GDEPT) is a superior gene therapy method when compared to chemotherapy and radiotherapy procedures, proven to be effective against many types of cancer in pre-clinical evaluations and clinical trials. Gene therapy that utilizes a single enzyme/prodrug combination targeting a single cellular mechanism needs significant overexpression of delivered therapeutic gene in order to achieve therapy response. Hence, to overcome this obstacle we recently developed a dual therapeutic reporter gene fusion that uses two different prodrugs, targeting two distinct cellular mechanisms in order to achieve effective therapy with a limited expression of delivered transgenes. In addition, imaging therapeutic reporter genes offers additional information that indirectly correlates gene delivery, expression, and functional effectiveness as a theranostic approach. In the present study, we evaluate the therapeutic potential of HSV1-sr39TK-NTR fusion dual suicide gene therapy system that we recently developed, in MDA-MB-231 triple negative breast cancer lung-metastatic lesions in a mouse model. We compared the therapeutic potential of HSV1-sr39TK-NTR fusion with respective dual prodrugs GCV-CB1954 with HSV1-sr39TK/GCV and NTR/CB1954 single enzyme prodrug system in this highly resistant metastatic lesion of the lungs. In vitro optimization of dose and duration of exposure to GCV and CB1954 was performed in MDA-MB-231 cells. Drug combinations of 1 μg/ml GCV and 10 μM CB1954 for 3 days was found to be optimal regimen for induction of significant cell death, as assessed by FACS analysis. In vivo therapeutic evaluation in animal models showed a complete ablation of lung metastatic nodules of MDA-MB-231 triple negative breast cancer cells following

  10. MS/MS analysis and imaging of lipids across Drosophila brain using secondary ion mass spectrometry.

    Science.gov (United States)

    Phan, Nhu T N; Munem, Marwa; Ewing, Andrew G; Fletcher, John S

    2017-06-01

    Lipids are abundant biomolecules performing central roles to maintain proper functioning of cells and biological bodies. Due to their highly complex composition, it is critical to obtain information of lipid structures in order to identify particular lipids which are relevant for a biological process or metabolic pathway under study. Among currently available molecular identification techniques, MS/MS in secondary ion mass spectrometry (SIMS) imaging has been of high interest in the bioanalytical community as it allows visualization of intact molecules in biological samples as well as elucidation of their chemical structures. However, there have been few applications using SIMS and MS/MS owing to instrumental challenges for this capability. We performed MS and MS/MS imaging to study the lipid structures of Drosophila brain using the J105 and 40-keV Ar 4000 + gas cluster ion source, with the novelty being the use of MS/MS SIMS analysis of intact lipids in the fly brain. Glycerophospholipids were identified by MS/MS profiling. MS/MS was also used to characterize diglyceride fragment ions and to identify them as triacylglyceride fragments. Moreover, MS/MS imaging offers a unique possibility for detailed elucidation of biomolecular distribution with high accuracy based on the ion images of its fragments. This is particularly useful in the presence of interferences which disturb the interpretation of biomolecular localization. Graphical abstract MS/MS was performed during time-of-flight secondary ion mass spectrometry (ToF-SIMS) analysis of Drosophila melongaster (fruit fly) to elucidate the structure and origin of different chemical species in the brain including a range of different phospholipid classes (PC, PI, PE) and di- and triacylglycerides (DAG & TAG) species where reference MS/MS spectra provided a potential means of discriminating between the isobaric [M-OH] + ion of DAGs and the [M-RCO] + ion of TAGs.

  11. PENERAPAN PEMBELAJARAN TARI GANTAR UNTUK MENINGKATKAN MOTORIK KASAR ANAK USIA DINI DENGAN MENGGUNAKAN KONSEP Developmentally Appropriate Practice (DAP DI TK KARTIKA V-66 BALIKPAPANTAHUN PELAJARAN 2014-2015

    Directory of Open Access Journals (Sweden)

    Euis Yuniastuti

    2015-12-01

    Full Text Available This research aimed at improving gross motor skills of Group B preschoolers through the learning of Tari Gantar traditional dance at TK Kartika V-66 Balikpapan using the concept of Developmentally Appropriate Practice (DAP in academic year 2014-2015. This research was designed as classroom action research which was carried out in three learning cycles: (1 action planning; (2 action execution; (3 observation and evaluation and (4 reflection. Researcher acted as dance teacher and researcher’s colleagues took a role as observers. The data was collected by filling in observation sheet, writing field notes, taking documentations and evaluating research activities. Qualitative analysis was equally performed to interpret the improvement of teacher’s and students’ activities in each cycle, while quantitative analysis was performed to explain the improvement of the students’ gross motor skills. Three aspects in gross motor skills measured in the research are locomotor, non-locomotor and manipulative skills. Results of the research exhibit the significant improvement of students’ gross motor skills throughout learning cycles. In cycle 1, the average value of students’ gross motor skills was about 75.13 while the average accomplishment percentage reached only 60.00%. In cycle 2, the average value of students’ gross motor skills increased at 76.31, while the average accomplishment percentage reached 80.00%.In cycle 3, the average value of students’ gross motor skills increased at 77.68while the average accomplishment percentage finally reached 92.00%.Teacher’s activity was significantly improved from cycle 1 at 70.00%, tocycle2at 86.67 % and finally to cycle 3at 100.00%.Finally, students’ activity was significantly improved from cycle 1 at 46.67%, to cycle2at 73.33% and finally to cycle 3 at 86.67%.

  12. MS/MS of synthetic peptide is not sufficient to confirm new types of protein modifications.

    Science.gov (United States)

    Lee, Sangkyu; Tan, Minjia; Dai, Lunzhi; Kwon, Oh Kwang; Yang, Jeong Soo; Zhao, Yingming; Chen, Yue

    2013-02-01

    Protein post-translational modification (PTM) is one of the major regulatory mechanisms that fine-tune protein functions. Undescribed mass shifts, which may suggest novel types of PTMs, continue to be discovered because of the availabilities of more sensitive mass spectrometry technologies and more powerful sequence alignment algorithms. In this study, the histone extracted from HeLa cells was analyzed using an approach that takes advantages of in vitro propionylation, efficient peptide separation using isoelectric focusing fractionation, and the high sensitivity of the linear ion trap coupled with hybrid FT mass spectrometer. One modified peptide was identified with a new type of protein modification (+42 Da), which was assigned to acetylation of threonine 15 in histone2A. The modified peptide was verified by careful manual evaluation of the tandem mass spectrum and confirmed by high-resolution MS/MS analysis of the corresponding synthetic peptide. However, HPLC coelution and MS/MS/MS of key ions showed that the +42 Da mass shifts at threonine residue did not correspond to acetylation. The key fragment ion, y4, in the MS/MS/MS spectra (indicative of the modification site) differed between the in vivo and synthetic peptide. We showed that the misidentification was originated from sequence homologues and chemical derivitization during sample preparation. This result indicated that a more stringent procedure that includes MS/MS, MS/MS/MS, and HPLC coelution of synthetic peptides is required to identify a new PTM.

  13. Analytical Method Details (MS): SE7_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE7_MS1 LC-Orbitrap-MS ESI positive method 1 Agilent1100 HPLC (Agilent), LTQ-Orbitr...s injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent),

  14. Analytical Method Details (MS): SE6_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE6_MS1 LC-Orbitrap-MS ESI positive method 1 Agilent1100 HPLC (Agilent), LTQ-Orbitr...s injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent),

  15. Structure Elucidation of Unknown Metabolites in Metabolomics by Combined NMR and MS/MS Prediction.

    Science.gov (United States)

    Boiteau, Rene M; Hoyt, David W; Nicora, Carrie D; Kinmonth-Schultz, Hannah A; Ward, Joy K; Bingol, Kerem

    2018-01-17

    We introduce a cheminformatics approach that combines highly selective and orthogonal structure elucidation parameters; accurate mass, MS/MS (MS²), and NMR into a single analysis platform to accurately identify unknown metabolites in untargeted studies. The approach starts with an unknown LC-MS feature, and then combines the experimental MS/MS and NMR information of the unknown to effectively filter out the false positive candidate structures based on their predicted MS/MS and NMR spectra. We demonstrate the approach on a model mixture, and then we identify an uncatalogued secondary metabolite in Arabidopsis thaliana . The NMR/MS² approach is well suited to the discovery of new metabolites in plant extracts, microbes, soils, dissolved organic matter, food extracts, biofuels, and biomedical samples, facilitating the identification of metabolites that are not present in experimental NMR and MS metabolomics databases.

  16. Application of LC–MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids

    OpenAIRE

    Haneef, Jamshed; Shaharyar, Mohammad; Husain, Asif; Rashid, Mohd; Mishra, Ravinesh; Parveen, Shama; Ahmed, Niyaz; Pal, Manoj; Kumar, Deepak

    2013-01-01

    Liquid chromatography tandem mass chromatography (LCâMS/MS) is an important hyphenated technique for quantitative analysis of drugs in biological fluids. Because of high sensitivity and selectivity, LCâMS/MS has been used for pharmacokinetic studies, metabolites identification in the plasma and urine. This manuscript gives comprehensive analytical review, focusing on chromatographic separation approaches (column packing materials, column length and mobile phase) as well as different acquisiti...

  17. Application of LC–MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids

    OpenAIRE

    Haneef, Jamshed; Shaharyar, Mohammad; Husain, Asif; Rashid, Mohd; Mishra, Ravinesh; Parveen, Shama; Ahmed, Niyaz; Pal, Manoj; Kumar, Deepak

    2013-01-01

    Liquid chromatography tandem mass chromatography (LC–MS/MS) is an important hyphenated technique for quantitative analysis of drugs in biological fluids. Because of high sensitivity and selectivity, LC–MS/MS has been used for pharmacokinetic studies, metabolites identification in the plasma and urine. This manuscript gives comprehensive analytical review, focusing on chromatographic separation approaches (column packing materials, column length and mobile phase) as well as different acquisiti...

  18. Analytical Method Details (MS): SE48_MS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE48_MS01 LC–QTOF-MS/MS analysis Waters ACQUITY UPLC System and Waters Xevo G2 QTOF mass spectrometer...tor in the range of 200–600 nm. The sample injection volume was 1μL. MS detection was performed on a Waters Xevo G2 QTOF mass spectro...meter with an electrospray ionization (ESI) interface (W

  19. Metabolomics data normalization with EigenMS.

    Directory of Open Access Journals (Sweden)

    Yuliya V Karpievitch

    Full Text Available Liquid chromatography mass spectrometry has become one of the analytical platforms of choice for metabolomics studies. However, LC-MS metabolomics data can suffer from the effects of various systematic biases. These include batch effects, day-to-day variations in instrument performance, signal intensity loss due to time-dependent effects of the LC column performance, accumulation of contaminants in the MS ion source and MS sensitivity among others. In this study we aimed to test a singular value decomposition-based method, called EigenMS, for normalization of metabolomics data. We analyzed a clinical human dataset where LC-MS serum metabolomics data and physiological measurements were collected from thirty nine healthy subjects and forty with type 2 diabetes and applied EigenMS to detect and correct for any systematic bias. EigenMS works in several stages. First, EigenMS preserves the treatment group differences in the metabolomics data by estimating treatment effects with an ANOVA model (multiple fixed effects can be estimated. Singular value decomposition of the residuals matrix is then used to determine bias trends in the data. The number of bias trends is then estimated via a permutation test and the effects of the bias trends are eliminated. EigenMS removed bias of unknown complexity from the LC-MS metabolomics data, allowing for increased sensitivity in differential analysis. Moreover, normalized samples better correlated with both other normalized samples and corresponding physiological data, such as blood glucose level, glycated haemoglobin, exercise central augmentation pressure normalized to heart rate of 75, and total cholesterol. We were able to report 2578 discriminatory metabolite peaks in the normalized data (p<0.05 as compared to only 1840 metabolite signals in the raw data. Our results support the use of singular value decomposition-based normalization for metabolomics data.

  20. Integrity assessment report of tanks TK-101 and TK-102

    International Nuclear Information System (INIS)

    MCSHANE, D.S.

    1999-01-01

    This Integrity Assessment Report (IAR) is prepared by Fluor Daniel Northwest (FDNW) for Waste Management Federal Services of Hanford, Inc., (WMH), the operations contractor; Fluor Daniel Hanford (FDH), the Hanford Site Manager; and the U. S. Department of Energy (DOE), the system owner. This IAR addresses the evaluation of Tanks 101 and 102 and other existing components located in the 219-S Waste Handling Facility. This report will be included in the Part B Permit for the 2226 Laboratory and is a portion of the integrity assessment of the overall 222-5 Laboratory radioactive liquid waste disposal system. This IAR is prepared in accordance with WAC 173-303, Dangerous Waste Regulations; Section 640(2), ''Assessment of Existing Tank Systems Integrity .''

  1. The therapeutic effect of PEI-Mn0.5Zn0.5Fe2O4 nanoparticles/pEgr1-HSV-TK/GCV associated with radiation and magnet-induced heating on hepatoma

    Science.gov (United States)

    Lin, Mei; Huang, Junxing; Zhang, Jia; Wang, Li; Xiao, Wei; Yu, Hong; Li, Yuntao; Li, Hongbo; Yuan, Chenyan; Hou, Xinxin; Zhang, Hao; Zhang, Dongsheng

    2013-01-01

    Comprehensive therapy based on the integration of hyperthermia, radiation, gene therapy and chemotherapy is a promising area of study in cancer treatment. Using PEI-Mn0.5Zn0.5Fe2O4 nanoparticles (PEI-MZF-NPs) as a gene transfer vector, the authors transfected self-prepared pEgr1-HSV-TK into HepG2 cells and measured the expression of the exogenous gene HSV-TK by RT-PCR. The results showed that HSV-TK was successfully transfected into HepG2 cells and the expression levels of HSV-TK remained stable. Besides, PEI-MZF-NPs were used as magnetic media for thermotherapy to treat hepatoma by magnet-induced heating, combined with radiation-gene therapy. Both in vitro and in vivo results suggest that this combined treatment with gene, radiation and heating has a better therapeutic effect than any of them alone. The apoptotic rate and necrotic rate of the combined treatment group was 51.84% and 15.45%, respectively. In contrast, it was only 20.55% and 6.80% in the radiation-gene group, 7.49% and 3.62% in the radiation-alone group, 15.23% and 7.90% in the heating-alone group, and only 3.52% and 2.16% in the blank control group. The inhibition rate of cell proliferation (88.5%) of the combined treatment group was significantly higher than that of the radiation-gene group (59.5%), radiation-alone group (37.6%) and heating-alone group (60.6%). The tumor volume and mass inhibition rate of the combined treatment group was 94.45% and 93.38%, respectively, significantly higher than 41.28% and 33.58% of the radiation-alone group, 60.76% and 52.18% of the radiation-gene group, 79.91% and 77.40% of the heating-alone group. It is therefore concluded that this combined application of heating, radiation and gene therapy has a good synergistic and complementary effect and PEI-MZF-NPs can act as a novel non-viral gene vector and magnetic induction medium, which offers a viable approach for the treatment of cancer.

  2. Analytical Method Details (MS): SE53_MS03 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE53_MS03 CE-TOF/MS Agilent CE capillary electrophoresis system,Agilent G3250AA LC/MSD TOF system,Agile...nt 1100 series binary HPLC pump,G1603A Agilent CE-MS adapter and G1607A Agilent CE-ESI-M...s were performed using an Agilent CE capillary electrophoresis system (Agilent Technologies, Waldbronn, Germany), an Agile...nt G3250AA LC/MSD TOF system (Agilent Technologies, Palo Alto, CA), an Agilent 1100 series bin...ary HPLC pump, and the G1603A Agilent CE-MS adapter and G1607A Agilent CE-ESI-MS

  3. A comparison of direct infusion MS and GC-MS for metabolic footprinting of yeast mutants

    DEFF Research Database (Denmark)

    Mass, S.; Villas-Bôas, Silas Granato; Hansen, Michael Adsetts Edberg

    2007-01-01

    -deletion mutants. Filtered fermentation broth samples were analyzed by GC-MS and direct infusion ESI-MS. The potential of both methods in producing specific and, therefore, discriminant metabolite profiles was evaluated using samples from several yeast deletion mutants grown in batch-culture conditions....... Thus, the GC-MS method is good for classification of mutants with altered nitrogen regulation as it primarily measures amino acids, whereas this method cannot classify mutants involved in regulation of phospholipids metabolism as well as the direct infusion MS (DI-MS) method. From the analysis, we find...

  4. The neurophysiologist perspective into MS plasticity

    Directory of Open Access Journals (Sweden)

    Elise eHoudayer

    2015-09-01

    Full Text Available Multiple sclerosis (MS is a frequent, highly debilitating inflammatory demyelinating disease, starting to manifest in early adulthood and presenting a wide variety of symptoms which are often resistant to pharmacological treatments. Cortical dysfunctions have been demonstrated to be key components of MS condition, and plasticity of the corticospinal motor system is highly involved in major MS symptoms, such as fatigue, spasticity or pain. Cortical dysfunction in MS can be studied with neurophysiological tools such as electroencephalography (EEG and related techniques (evoked potentials – EPs or transcranial magnetic stimulation (TMS. These techniques are now widely used to provide essential elements of MS diagnosis and can also be used to modulate plasticity. Indeed the recent development of non-invasive brain stimulation (NIBS techniques able to induce cortical plasticity, such as repetitive TMS or transcranial direct current stimulation (tDCS, has brought promising results as add-on treatments.In this review we will focus on the use of these tools (EEG, TMS to study plasticity in MS and on the major techniques used to modulate plasticity in MS.

  5. The Neurophysiologist Perspective into MS Plasticity.

    Science.gov (United States)

    Houdayer, Elise; Comi, Giancarlo; Leocani, Letizia

    2015-01-01

    Multiple sclerosis (MS) is a frequent, highly debilitating inflammatory demyelinating disease, starting to manifest in early adulthood and presenting a wide variety of symptoms, which are often resistant to pharmacological treatments. Cortical dysfunctions have been demonstrated to be key components of MS condition, and plasticity of the corticospinal motor system is highly involved in major MS symptoms, such as fatigue, spasticity, or pain. Cortical dysfunction in MS can be studied with neurophysiological tools, such as electroencephalography (EEG) and related techniques (evoked potentials) or transcranial magnetic stimulation (TMS). These techniques are now widely used to provide essential elements of MS diagnosis and can also be used to modulate plasticity. Indeed, the recent development of non-invasive brain stimulation techniques able to induce cortical plasticity, such as repetitive TMS or transcranial direct current stimulation, has brought promising results as add-on treatments. In this review, we will focus on the use of these tools (EEG and TMS) to study plasticity in MS and on the major techniques used to modulate plasticity in MS.

  6. Venocentric lesions: an MRI marker of MS?

    Directory of Open Access Journals (Sweden)

    Matthew P. Quinn

    2013-07-01

    Full Text Available From the earliest descriptions of MS, the venocentric characteristic of plaques was noted. Recently, numerous MRI studies have proposed this finding as a prospective biomarker for MS, which might aid in differentiating MS from other diseases with similar MRI findings. High field MRI studies have shown that penetrating veins can be detected in most MS lesions using T2* weighted or susceptibility weighted imaging. Future studies must address the feasibility of imaging such veins in a clinically practical context. The specificity of this biomarker has been studied only in a limited capacity. Results in microangiopathic lesions are conflicting, whereas asymptomatic white matter hyperintensities as well as lesions of NMO are less frequently venocentric compared to MS plaques. Prospective studies have shown that the presence of venocentric lesions at an early clinical presentation is highly predictive of future MS diagnosis. This is very promising, but work remains to be done to confirm or exclude lesions of common MS mimics, such as ADEM, as venocentric. A number of technical challenges must be addressed before the introduction of this technique as a complementary tool in current diagnostic procedures.

  7. MS/MS networking guided analysis of molecule and gene cluster families

    Science.gov (United States)

    Nguyen, Don Duy; Wu, Cheng-Hsuan; Moree, Wilna J.; Lamsa, Anne; Medema, Marnix H.; Zhao, Xiling; Gavilan, Ronnie G.; Aparicio, Marystella; Atencio, Librada; Jackson, Chanaye; Ballesteros, Javier; Sanchez, Joel; Watrous, Jeramie D.; Phelan, Vanessa V.; van de Wiel, Corine; Kersten, Roland D.; Mehnaz, Samina; De Mot, René; Shank, Elizabeth A.; Charusanti, Pep; Nagarajan, Harish; Duggan, Brendan M.; Moore, Bradley S.; Bandeira, Nuno; Palsson, Bernhard Ø.; Pogliano, Kit; Gutiérrez, Marcelino; Dorrestein, Pieter C.

    2013-01-01

    The ability to correlate the production of specialized metabolites to the genetic capacity of the organism that produces such molecules has become an invaluable tool in aiding the discovery of biotechnologically applicable molecules. Here, we accomplish this task by matching molecular families with gene cluster families, making these correlations to 60 microbes at one time instead of connecting one molecule to one organism at a time, such as how it is traditionally done. We can correlate these families through the use of nanospray desorption electrospray ionization MS/MS, an ambient pressure MS technique, in conjunction with MS/MS networking and peptidogenomics. We matched the molecular families of peptide natural products produced by 42 bacilli and 18 pseudomonads through the generation of amino acid sequence tags from MS/MS data of specific clusters found in the MS/MS network. These sequence tags were then linked to biosynthetic gene clusters in publicly accessible genomes, providing us with the ability to link particular molecules with the genes that produced them. As an example of its use, this approach was applied to two unsequenced Pseudoalteromonas species, leading to the discovery of the gene cluster for a molecular family, the bromoalterochromides, in the previously sequenced strain P. piscicida JCM 20779T. The approach itself is not limited to 60 related strains, because spectral networking can be readily adopted to look at molecular family–gene cluster families of hundreds or more diverse organisms in one single MS/MS network. PMID:23798442

  8. In Vivo Metabolism Study of Xiamenmycin A in Mouse Plasma by UPLC-QTOF-MS and LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Feng Lei

    2015-01-01

    Full Text Available Xiamenmycin A is an antifibrotic leading compound with a benzopyran skeleton that is isolated from mangrove-derived Streptomyces xiamenensis. As a promising small molecule for fibrotic diseases, less information is known about its metabolic characteristics in vivo. In this study, the time-course of xiamenmycin A in mouse plasma was investigated by relative quantification. After two types of administration of xiamenmycin A at a single dose of 10 mg/kg, the plasma concentrations were measured quantitatively by LC-MS/MS. The dynamic changes in the xiamenmycin A concentration showed rapid absorption and quick elimination in plasma post-administration. Four metabolites (M1–M4 were identified in blood by UPLC-QTOF-MS, and xiamenmycin B (M3 is the principal metabolite in vivo, as verified by comparison of the authentic standard sample. The structures of other metabolites were identified based on the characteristics of their MS and MS/MS data. The newly identified metabolites are useful for understanding the metabolism of xiamenmycin A in vivo, aiming at the development of an anti-fibrotic drug candidate for the therapeutic treatment of excessive fibrotic diseases.

  9. Analysis of Thiodiglycol: Validation of Semi-Volatile Analysis by HPLC-MS/MS by EPA Method MS777

    Energy Technology Data Exchange (ETDEWEB)

    Owens, J; Koester, C

    2008-07-24

    The Environmental Protection Agency's (EPA) Region 5 Chicago Regional Laboratory (CRL) developed a method for the analysis of thiodiglycol, the breakdown product of the sulfur mustard HD, in water by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS), titled Method EPA MS777 (hereafter referred to as EPA CRL SOP MS777). This draft standard operating procedure (SOP) was distributed to multiple EPA laboratories and to Lawrence Livermore National Laboratory, which was tasked to serve as a reference laboratory for EPA's Environmental Reference Laboratory Network (ERLN) and to develop and validate analytical procedures. The primary objective of this study was to verify the analytical procedures described in MS777 for analysis of thiodiglycol in aqueous samples. The gathered data from this study will be used to: (1) demonstrate analytical method performance; (2) generate quality control acceptance criteria; and (3) revise the SOP to provide a validated method that would be available for use during a homeland security event. The data contained in this report will be compiled, by EPA CRL, with data generated by other EPA Regional laboratories so that performance metrics of Method EPA MS777 can be determined.

  10. Significance of MRI Perivascular Spaces in MS

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2008-10-01

    Full Text Available The role of perivascular Virchow-Robin spaces was investigated in 45 multiple sclerosis (MS patients and 30 healthy controls, in a study at Charite-Universitaetsmedizin Berlin, and Goethe University, Frankfurt.

  11. The Importance of “Orff-Schulwerk” for Musical Social-Integrative Pedagogy and Music Therapy (English translation: Gloria Litwin

    Directory of Open Access Journals (Sweden)

    Karin Schumacher

    2013-12-01

    Full Text Available Which features of Carl Orff and Gunild Keetman’s ideas are relevant for work in the community and therapy? A child who is mentally or physically handicapped, emotionally or sensorially disabled or on the autistic spectrum, is never just only that. The characteristics of a person that are not expressed in these terms, however, are precisely the ones that make musical communication and thus a connection with so-called ‘normal’ people possible. A historical review will demonstrate the sources from which the adaptation of the Schulwerk for work with handicapped and disturbed children and young people was made possible and meaningful. A brief section will define the fields of “Music Education”, “Music in Special Needs and Community” and “Music Therapy” in order to distinguish them and highlight the contents they have in common. The author uses quotes by Carl Orff in order to document the basis of her ideas for therapeutic work and describes how pedagogues and therapists, from the early sixties until today, have developed them for and together with different groups with special needs: - The multi-sensorial approach through speech, free and bound rhythm, movement, singing and playing instruments provides possibilities for spontaneous creative play in a social context, even if one important sensorial area is severely damaged. - Every member of an integrative music and movement group is participating actively in a creative process. - The instrumentarium allows participants to play together in a spontaneous way. - Relationships developed through musical expression and play as an encounter between two people forms the basis for emotional development. - Musical reception and expression is independent of intellectual capacity, age and previous musical experience.

  12. High-speed MALDI MS/MS imaging mass spectrometry using continuous raster sampling.

    Science.gov (United States)

    Prentice, Boone M; Chumbley, Chad W; Caprioli, Richard M

    2015-04-01

    A matrix-assisted laser desorption/ionization time of flight/time of flight tandem mass spectrometer (MALDI TOF/TOF) has been used for high-speed precursor/fragment ion transition image acquisition. High-throughput analysis is facilitated by an Nd:YLF solid state laser capable of pulse repetition rates up to 5 kHz, a high digitizer acquisition rate (up to 50 pixels/s), and continuous laser raster sampling. MS/MS experiments are enabled through the use of a precision timed ion selector, second source acceleration, and a dedicated collision cell. Continuous raster sampling is shown here to facilitate rapid MS/MS ion image acquisition from thin tissue sections for the drug rifampicin and for a common kidney lipid, SM4s(d18:1/24:1). The ability to confirm the structural identity of an analyte as part of the MS/MS imaging experiment is an essential part of the analysis. Additionally, the increase in sensitivity and specificity afforded by an MS/MS approach is highly advantageous, especially when interrogating complex chemical environments such as those in biological tissues. Herein, we report continuous laser raster sampling TOF/TOF imaging methodologies which demonstrate 8 to 14-fold increases in throughput compared with existing MS/MS instrumentation, an important advantage when imaging large areas on tissues. Copyright © 2015 John Wiley & Sons, Ltd.

  13. Simultaneous analysis of 17 diuretics in dietary supplements by HPLC and LC-MS/MS.

    Science.gov (United States)

    Woo, H; Kim, J W; Han, K M; Lee, J H; Hwang, I S; Lee, J H; Kim, J; Kweon, S J; Cho, S; Chae, K R; Han, S Y; Kim, J

    2013-01-01

    In order to test health foods for illegally added diuretics for weight loss, we developed simple, rapid, selective, and sensitive methods using HPLC and LC-MS/MS for the simultaneous analysis of 17 diuretics in dietary supplements. HPLC conditions were set with a Capcell-pak C18, using a mobile phase consisting of gradient conditions, UV detection at 254 nm and validated for linearity (r(2)> 0.999), precision (CV ≤ 3%), recoveries (90.4-102.8%) and reproducibility. Identification and quantification of 17 diuretics were accomplished by ion-spray LC-MS/MS using multiple reaction monitoring (MRM). The chromatographic separation was carried out under the reversed-phase mechanism on an HSS-T3 column. The LC-MS/MS method was validated for linearity (r(2)> 0.99) and precision (CV Diuretics were not detected in all samples. Extraction recovery was also investigated and the extraction recoveries in different formulations were from 88% to 110% and from 81% to 116% using HPLC and LC-MS/MS, respectively. There was no significant difference in recoveries in the type of dietary supplements. Based on this result, the developed methods to monitor illegal drug adulterations in dietary supplements using HPLC and LC-MS/MS are simple, fast and reliable. Therefore, it is applicable to routine drug-adulteration screening.

  14. Multi-Segment Direct Inject nano-ESI-LTQ-FT-ICR-MS/MS For Protein Identification

    Directory of Open Access Journals (Sweden)

    Neal Rachel E

    2011-07-01

    Full Text Available Abstract Reversed phase high performance liquid chromatography (HPLC interfaced to electrospray tandem mass spectrometry (MS/MS is commonly used for the identification of peptides from proteolytically cleaved proteins embedded in a polyacrylamide gel matrix as well as for metabolomics screening. HPLC separations are time consuming (30-60 min average, costly (columns and mobile phase reagents, and carry the risk of column carry over between samples. The use of a chip-based nano-ESI platform (Advion NanoMate based on replaceable nano-tips for sample introduction eliminates sample cross-contamination, provides unchanging sample matrix, and enhances spray stability with attendant increases in reproducibility. Recent papers have established direct infusion nano-ESI-MS/MS utilizing the NanoMate for protein identification of gel spots based on full range MS scans with data dependent MS/MS. In a full range scan, discontinuous ion suppression due to sample matrix can impair identification of putative mass features of interest in both the proteomic and metabolomic workflows. In the current study, an extension of an established direct inject nano-ESI-MS/MS method is described that utilizes the mass filtering capability of an ion-trap for ion packet separation into four narrow mass ranges (50 amu overlap with segment specific dynamic data dependent peak inclusion for MS/MS fragmentation (total acquisition time of 3 minutes. Comparison of this method with a more traditional nanoLC-MS/MS based protocol utilizing solvent/sample stream splitting to achieve nanoflow demonstrated comparable results for protein identification from polyacrylamide gel matrices. The advantages of this method include full automation, lack of cross-contamination, low cost, and high throughput.

  15. Expert System for Computer-assisted Annotation of MS/MS Spectra*

    Science.gov (United States)

    Neuhauser, Nadin; Michalski, Annette; Cox, Jürgen; Mann, Matthias

    2012-01-01

    An important step in mass spectrometry (MS)-based proteomics is the identification of peptides by their fragment spectra. Regardless of the identification score achieved, almost all tandem-MS (MS/MS) spectra contain remaining peaks that are not assigned by the search engine. These peaks may be explainable by human experts but the scale of modern proteomics experiments makes this impractical. In computer science, Expert Systems are a mature technology to implement a list of rules generated by interviews with practitioners. We here develop such an Expert System, making use of literature knowledge as well as a large body of high mass accuracy and pure fragmentation spectra. Interestingly, we find that even with high mass accuracy data, rule sets can quickly become too complex, leading to over-annotation. Therefore we establish a rigorous false discovery rate, calculated by random insertion of peaks from a large collection of other MS/MS spectra, and use it to develop an optimized knowledge base. This rule set correctly annotates almost all peaks of medium or high abundance. For high resolution HCD data, median intensity coverage of fragment peaks in MS/MS spectra increases from 58% by search engine annotation alone to 86%. The resulting annotation performance surpasses a human expert, especially on complex spectra such as those of larger phosphorylated peptides. Our system is also applicable to high resolution collision-induced dissociation data. It is available both as a part of MaxQuant and via a webserver that only requires an MS/MS spectrum and the corresponding peptides sequence, and which outputs publication quality, annotated MS/MS spectra (www.biochem.mpg.de/mann/tools/). It provides expert knowledge to beginners in the field of MS-based proteomics and helps advanced users to focus on unusual and possibly novel types of fragment ions. PMID:22888147

  16. Multiple structure alignment with msTALI.

    Science.gov (United States)

    Shealy, Paul; Valafar, Homayoun

    2012-05-20

    Multiple structure alignments have received increasing attention in recent years as an alternative to multiple sequence alignments. Although multiple structure alignment algorithms can potentially be applied to a number of problems, they have primarily been used for protein core identification. A method that is capable of solving a variety of problems using structure comparison is still absent. Here we introduce a program msTALI for aligning multiple protein structures. Our algorithm uses several informative features to guide its alignments: torsion angles, backbone Cα atom positions, secondary structure, residue type, surface accessibility, and properties of nearby atoms. The algorithm allows the user to weight the types of information used to generate the alignment, which expands its utility to a wide variety of problems. msTALI exhibits competitive results on 824 families from the Homstrad and SABmark databases when compared to Matt and Mustang. We also demonstrate success at building a database of protein cores using 341 randomly selected CATH domains and highlight the contribution of msTALI compared to the CATH classifications. Finally, we present an example applying msTALI to the problem of detecting hinges in a protein undergoing rigid-body motion. msTALI is an effective algorithm for multiple structure alignment. In addition to its performance on standard comparison databases, it utilizes clear, informative features, allowing further customization for domain-specific applications. The C++ source code for msTALI is available for Linux on the web at http://ifestos.cse.sc.edu/mstali.

  17. Application of LC/MS/MS Techniques to Development of US ...

    Science.gov (United States)

    This presentation will describe the U.S. EPA’s drinking water and ambient water method development program in relation to the process employed and the typical challenges encountered in developing standardized LC/MS/MS methods for chemicals of emerging concern. The EPA’s Drinking Water Contaminant Candidate List and Unregulated Contaminant Monitoring Regulations, which are the driving forces behind drinking water method development, will be introduced. Three drinking water LC/MS/MS methods (Methods 537, 544 and a new method for nonylphenol) and two ambient water LC/MS/MS methods for cyanotoxins will be described that highlight some of the challenges encountered during development of these methods. This presentation will provide the audience with basic understanding of EPA's drinking water method development program and an introduction to two new ambient water EPA methods.

  18. Sulphur fertilization influences the sulphur species composition in Allium sativum: sulphomics using HPLC-ICPMS/MS-ESI-MS/MS.

    Science.gov (United States)

    Raab, Andrea; Ronzan, Marilena; Feldmann, Joerg

    2017-10-18

    Garlic (A. sativum) contains a large number of small sulphur (S)-containing metabolites, which are important for its taste and smell and vary with A. sativum variety and growth conditions. This study was designed to investigate the influence of different sulphur-fertilization regimes on low molecular weight S-species by attempting the first sulphur mass balance in A. sativum roots and bulbs using HPLC-ICPMS/MS-ESI-MS/MS. Species unspecific quantification of acid soluble S-containing metabolites was achieved using HPLC-ICP-MS/MS. For identification of the compounds, high resolution ESI-MS (Orbitrap LTQ and q-TOF) was used. The plants contained up to 54 separated sulphur-containing compounds, which constitute about 80% of the total sulphur present in A. sativum. The roots and bulbs of A. sativum contained the same compounds, but not necessarily the same amounts and proportions. The S-containing metabolites in the roots reacted more sensitively to manipulations of sulphur fertilization than those compounds in the bulbs. In addition to known compounds (e.g. γ-glutamyl-S-1-propenylcysteine) we were able to identify and partially quantify 31 compounds. Three as yet undescribed S-containing compounds were also identified and quantified for the first time. Putative structures were assigned to the oxidised forms of S-1-propenylmercaptoglutathione, S-2-propenylmercaptoglutathione, S-allyl/propenyl-containing PC-2 and 2-amino-3-[(2-carboxypropyl)sulfanyl]propanoic acid. The parallel use of ICP-MS/MS as a sulphur-specific detector and ESI-MS as a molecular detector simplifies the identification and quantification of sulphur containing metabolites without species specific standards. This non-target analysis approach enables a mass balance approach and identifies the occurrence of the so far unidentified organosulphur compounds. The experiments showed that the sulphur-fertilization regime does not influence sulphur-speciation, but the concentration of some S

  19. Conference report: 60th American Society for MS Conference on MS and Allied Topics.

    Science.gov (United States)

    Touboul, David; Seyer, Alexandre; Becher, François

    2012-09-01

    The 60th American Society for Mass Spectrometry (ASMS) Conference on MS and Allied Topics was held in May 2012 at Vancouver in Canada. This international congress is the largest forum exclusively dedicated to MS through its diverse aspects: fundamental, method development and applications to biomolecules. This 4-day conference is also highly appreciated for discussion with the MS manufacturers presenting their latest instrumental developments.

  20. Novel LC-MS/MS method for plasma vancomycin: comparison with immunoassays and clinical impact.

    Science.gov (United States)

    Oyaert, Matthijs; Peersman, Nele; Kieffer, Davy; Deiteren, Kathleen; Smits, Anne; Allegaert, Karel; Spriet, Isabel; Van Eldere, Johan; Verhaegen, Jan; Vermeersch, Pieter; Pauwels, Steven

    2015-02-20

    Accurate quantification of vancomycin in plasma is important for adequate dose-adjustment. As literature suggests between-method differences, our first objective was to develop a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for total vancomycin in human plasma and to compare frequently used immunoassays with this method. Secondly, we investigated the clinical impact of between-method quantification differences. For LC-MS/MS, lithium heparin plasma was extracted by adding a precipitation reagent containing the internal standard (vancomycin-des-leucine). Analysis was performed on an Acquity TQD mass spectrometer equipped with an Acquity UPLC 2795 separations module. Our method was analytically validated and compared with four frequently used immunoassays from four different manufacturers. Vancomycin concentrations were clinically classified as toxic, therapeutic and sub-therapeutic. Clinical discordance was calculated using LC-MS/MS as a reference. A novel LC-MS/MS method using protein precipitation as sole pretreatment and an analysis time of 5.0 min was developed. The assay had a total imprecision of 2.6-8.5%, a limit of quantification of 0.3 mg/L and an accuracy ranging from 101.4 to 111.2%. Using LC-MS/MS as reference, three immunoassays showed a mean proportional difference within 10% and one showed a substantial mean proportional difference of >20%. Clinical discordant interpretation of the obtained concentrations ranged from 6.1 to 22.2%. We developed a novel LC-MS/MS method for rapid analysis of total vancomycin concentrations in human plasma. Correlation of the method with immunoassays showed a mean proportional difference >20% for one of the assays, causing discordant clinical interpretation in more than 1 out of 5 samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Sulfur analysis of household briquettes using MS and GC/MS detection systems after reductive pyrolysis

    OpenAIRE

    Marinov, S. P.; Stefanova, M.; Stamenova, V.; Gonsalvesh, L.; CARLEER, Robert; YPERMAN, Jan

    2006-01-01

    The aim of the present study is to obtain more information concerning organic sulphur pollutants produced by some commercial briquettes based on coal and biomass. Reductive pyrolysis technique (AP-TPR), coupled "on-line" with mass spectrometer (MS) and "off-line" with GC/MS was used for qualitative and quantitative study of sulphur compounds emitted from household briquettes. By AP-TPR-GC/MS and inner standard application, prominent sulphur containing compounds were quantitatively determined....

  2. LC-MS-MS identification of drug metabolites obtained by metalloporphyrin mediated oxidation

    Directory of Open Access Journals (Sweden)

    Maurin Andrea J. M.

    2003-01-01

    Full Text Available In this paper we report the application of liquid chromatography-mass spectrometry (LC-MS-MS to the identification of the products formed by oxidation of albendazole and disopyramide with metalloporphyrins in dichloroethane, using iodosylbenzene as an oxygen donor. Our results show that LC-MS-MS is a powerful tool to study the in vitro metabolism of drugs, allowing the identification of known and unknown metabolites. In addition, it was observed that the catalyst system used resulted in the formation of the same metabolites as obtained in vivo, although for disopyramide other products were also observed.

  3. Hearing Status Survey in Define MS Patients

    Directory of Open Access Journals (Sweden)

    Davood Rostamian

    2003-08-01

    Full Text Available Objective: This study was aimed to survey the hearing status of the patients with definite MS. The results of this study may help to confirm the diagnosis and to improve our knowledge of different aspects of this disease and to optimize the treatment and rehabilitative techniques. Material & Methods: This study is performed in audiology clinic of rehabilitation science faculty of Iran medical science university as an assesive-analytic procedure in winter of 2000 in 40 MS patients (Referred from the MS clinic of Shohada Hospital of 20 to 45 years old. All the patients have no history of otologic problems. The results of conventional audiologist tests (PTA, SRT, SDS, imittance audiometry and ABR findings of MS patients were compared to the results of control group. Results: The differences between the means of low, high and mid frequency PTAve in MS patients and control group were statistically significant (P<0.05. There was no significant difference between two groups in the speech test's results, statistically. The results of ART test show statistically significant differences between experimental and control groups. The only differences in ABR results were seen for the absolute latency of the wave V and IPLs (I-V, III-V between two groups. The amplitude ratio of V/I in MS patients 1.5 was within the normal range (0.5 to 2. There was a statistically significant relationship between mean of mid frequency PTAve and prolongation of absolute latency of the wave V. The relationship between ABR and SDS and also between ART and ABR were not statistically significant. Finally, the results of this study suggested that low, high and mid frequency PTAve, ART, absolute latency of the wave V, and IPLs III-V, I-V can be used ascomplementory method to confirm the diagnosis of MS with other conventional methods such as: MRI, CSF analysis, SEP and VEP.

  4. Spatio-energetic cross-talk in photon counting detectors: Numerical detector model (PcTK) and workflow for CT image quality assessment.

    Science.gov (United States)

    Taguchi, Katsuyuki; Stierstorfer, Karl; Polster, Christoph; Lee, Okkyun; Kappler, Steffen

    2018-03-14

    The inter-pixel cross-talk of energy-sensitive photon counting x-ray detectors (PCDs) has been studied and an analytical model (version 2.1) has been developed for double-counting between neighboring pixels due to charge sharing and K-shell fluorescence x-ray emission followed by its re-absorption [Taguchi K, et al., Medical Physics 2016;43(12):6386-6404]. While the model version 2.1 simulated the spectral degradation well, it had the following problems that has been found to be significant recently: (1) The spectrum is inaccurate with smaller pixel sizes; (2) the charge cloud size must be smaller than the pixel size; (3) the model underestimates the spectrum/counts for 10-40 keV; and (4) the model version 2.1 cannot handle n-tuple-counting with n>2 (i.e., triple-counting or higher). These problems are inherent to the design of the model version 2.1; therefore, we developed a new model and addressed these problems in this study. We propose a new PCD cross-talk model (version 3.2; PcTK for "photon counting toolkit") that is based on a completely different design concept from the previous version. It uses a numerical approach and starts with a 2-D model of charge sharing (as opposed to an analytical approach and a 1-D model with version 2.1) and addresses all of the four problems. The model takes the following factors into account: (a) shift-variant electron density of the charge cloud (Gaussian-distributed), (b) detection efficiency, (c) interactions between photons and PCDs via photoelectric effect, and (d) electronic noise. Correlated noisy PCD data can be generated using either a multivariate normal number generator or a Poisson random number generator. The effect of the two parameters, the effective charge cloud diameter (d 0 ) and pixel size (d pix ), was studied and results were compared with Monte Carlo simulations and the previous model version 2.1. Finally, a script for the workflow for CT image quality assessment has been developed, which started with a

  5. MS in prose, poems and drama.

    Science.gov (United States)

    Riem, Philine; Karenberg, Axel

    2015-09-01

    Presentations of MS in fictional literature have not been previously researched. This paper surveys and analyses these portrayals of the disease for the first time. Relevant works in English and German were identified by means of keyword searches in online public access catalogues and search engines as well as old-fashioned research. The neurological and literary evaluation of these 7000 pages of text combines qualitative and quantitative methods. Between 1954 and 2012 at least 55 literary works appeared with an MS motif (35 novels, 18 poems, one novella and one drama). The authors were predominantly female and a third of them suffered from the disease. Patients in the novels largely reflect real epidemiology as regards symptoms and disease progression, while diagnostic and therapeutic options play a secondary role. From a literary point of view, 'entwicklungsromane', 'relationship novels' and 'young adult books' can be discerned. MS is often portrayed in metaphoric language as the enemy: a demon, an animalistic being, prison or an abyss. The MS motif evidences a medicalization of the literature as well as a literary portrayal of anthropological experiences. Well-written novels can contribute to the de-stigmatization of MS and impart basic medical knowledge. © The Author(s), 2015.

  6. An introduction to the microbiome and MS.

    Science.gov (United States)

    Trott, Sebastien; King, Irah L

    2018-01-01

    The human microbiota is composed of diverse forms of microorganisms that live on or in us and plays a crucial role in the health and development. Commensal species that reside in the intestine particularly influence host physiology at local and systemic levels. Multiple sclerosis (MS) is a debilitating autoimmune disorder of the central nervous system for which there is currently no cure. While the cause of MS is unknown, there is a growing body of evidence suggesting that the microbiota can play both pathogenic and protective roles in disease progression. In this review, we provide a brief overview, based on both animal and clinical studies, of the current understanding by which the microbiota may influence MS and discuss opportunities for therapeutic intervention that may alleviate the symptoms associated with this debilitating neuroimmunological disease.

  7. Analytical Method Details (MS): SE4_MS5 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE4_MS5 LC-FT-ICR-MS ESI positive method 5 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...uL sample is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agile

  8. Analytical Method Details (MS): SE4_MS2 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE4_MS2 LC-FT-ICR-MS ESI positive method 2 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...reatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100V (4.6 x 250 mm, 5 micrometer; T

  9. Analytical Method Details (MS): SE15_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE15_MS1 LC-Orbitrap-MS ESI positive method 1 Agilent1100 HPLC (Agilent), LTQ-Orbit... 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel ODS-100V (4

  10. Analytical Method Details (MS): SE9_MS2 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE9_MS2 LC-FT-ICR-MS ESI positive method 2 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100V (

  11. Analytical Method Details (MS): SE37_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE37_MS1 Metabolites extraction with 80% methanol and analysis by LC-Orbitrap-MS Agilent1100 HPLC (Agile...sis> A 10ul sample is injected into HPLC. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel ODS

  12. Analytical Method Details (MS): SE4_MS4 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE4_MS4 LC-FT-ICR-MS ESI positive method 4 Agilent1100 HPLC (Agilent), LTQ-FT (Ther... (Waters). 20uL sample is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agile...nt 1100 series (Agilent), Column: TSKgel-100V (4.6 x 250 mm, 5 micrometer; TOSOH),

  13. Analytical Method Details (MS): SE9_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE9_MS1 LC-FT-ICR-MS ESI positive method 1 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100V (

  14. Analytical Method Details (MS): SE2_MS3 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available 00.0-1500.0); 2: ITMS + c norm !corona !pi Dep MS/MS Most intense ion from (1)., Rejected mass=284.2000;300.2000;328.2000;344.2000;372.2000;388.2000;416.3000;432.3000;460.0000. ...

  15. Quantification of free and total sialic acid excretion by LC-MS/MS

    NARCIS (Netherlands)

    van der Ham, Maria; Prinsen, Berthil H. C. M. T.; Huijmans, Jan G. M.; Abeling, Nicolaas G. G. M.; Dorland, Bert; Berger, Ruud; de Koning, Tom J.; de Sain-van der Velden, Monique G. M.

    2007-01-01

    The main purpose for measuring urinary free sialic acid (FSA) is to diagnose sialic acid (SA) storage diseases. Elevated amounts of conjugated sialic acid (CSA) are observed in several diseases indicating the need to quantify CSA as well. A LC-MS/MS method for quantification of FSA and total sialic

  16. STS-47 MS Davis and MS Jemison with LBNP device in SLJ module aboard OV-105

    Science.gov (United States)

    1992-01-01

    STS-47 Mission Specialist (MS) N. Jan Davis (left) and MS Mae C. Jemison prepare the lower body negative pressure (LBNP) device for the LBNP experiment in the Spacelab Japan (SLJ) science module aboard the Earth-orbiting Endeavour, Orbiter Vehicle (OV) 105. Displayed on the aft end cone in the background is an Auburn University banner.

  17. Gluten Detection and Speciation by Liquid Chromatography Mass Spectrometry (LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Stephen Lock

    2013-12-01

    Full Text Available Liquid chromatography tandem mass spectrometry (LC-MS/MS has been used historically in proteomics research for over 20 years. However, until recently LC-MS/MS has only been routinely used in food testing for small molecule contaminant detection, for example pesticide and veterinary residue detection, and not as a replacement of microbiological food testing methods, specifically allergen analysis. Over the last couple of years, articles have started to be published which describe the detection of allergens by LC-MS/MS. In this article we will describe how LC-MS/MS can be applied in the area of gluten detection and how it can be used to specifically differentiate the species of gluten used in food, where specific markers for each variety of gluten can be simultaneously acquired and detected at the same time. The article will discuss the effect of variety on the peptide response observed from different wheat grain varieties and will describe the sample preparation protocol which is essential for generating the peptide markers used for speciation.

  18. Analytical Method Details (MS): SE53_MS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available lvent delay of 273 s. Data acquisition was performed on a Pegasus IV TOF mass spectrometer (LECO, St. Joseph...SE53_MS01 GC-TOF/MS Agilent 6890N gas chromatograph (Agilent Technologies) and Pegasus IV TOF mass spectrome...ter (LECO) EI Positive Each sample was ex

  19. Study on detection of drugs in slimming health foods using GC-MS/MS.

    Science.gov (United States)

    Yamamoto, Shinya; Sumioka, Shigeharu; Fujioka, Masanobu; Mikami, Eiichi; Miyamoto, Ken-ichi

    2011-01-01

    The determination of five drugs, fenfluramine (FEN), N-nitrosofenfluramine (NFE), sibutramine (SIB), mazindol (MAZ) and phenolphthalein (PHP), was studied in slimming health foods using GC-MS/MS. These drugs have been detected at high rates, especially in slimming health foods. Prolonged or excessive consumption of non-approved or unauthorized pharmaceuticals may cause serious adverse health consequences. In this study, samples were extracted with methanol and ultrasonication. Analyses were performed by GC-MS/MS, using established MS/MS parameters in the electron ionization (EI) mode and chemical ionization (CI) mode. In the EI mode, the recoveries of five drugs from several types of slimming health foods such as tablets, capsules and tea-bags spiked at 1 µg/mg (except PHP, spiked at 4 µg/mg) were in the range of 85.0-110.7% and 100 µg/mg (except PHP, spiked at 200 µg/mg) were 94.9-102.9%, respectively. In the CI mode, good recoveries of 80.3-102.2% (spiked at low concentration) and 92.8-103.2% (spiked at high concentration) were also obtained. We evaluated the present method using four slimming health foods, in which drugs had previously been detected. The results were similar to the previous results. These findings indicate that the present procedure for evaluating five drugs in slimming health foods by means of GC-MS/MS is useful.

  20. Increasing bioanalytical throughput using pcSFC-MS/MS: 10 minutes per 96-well plate.

    Science.gov (United States)

    Hoke, S H; Tomlinson, J A; Bolden, R D; Morand, K L; Pinkston, J D; Wehmeyer, K R

    2001-07-01

    The utility of packed-column supercritical, subcritical, and enhanced fluidity liquid chromatographies (pcSFC) for high-throughput applications has increased during the past few years. In contrast to traditional reversed-phase liquid chromatography, the addition of a volatile component to the mobile phase, such as CO2, produces a lower mobile-phase viscosity. This allows the use of higher flow rates which can translate into faster analysis times. In addition, the resulting mobile phase is considerably more volatile than the aqueous-based mobile phases that are typically used with LC-MS, allowing the entire effluent to be directed into the MS interface. High-throughput bioanalytical quantitation using pcSFC-MS/MS for pharmacokinetics applications is demonstrated in this report using dextromethorphan as a model compound. Plasma samples were prepared by automated liquid/liquid extraction in the 96-well format prior to pcSFC-MS/MS analysis. Three days of validation data are provided along with study sample data from a patient dosed with commercially available Vicks 44. Using pcSFC and MS/MS, dextromethorphan was quantified in 96-well plates at a rate of approximately 10 min/plate with average intraday accuracy of 9% or better. Daily relative standard deviations (RSDs) were less than 10% for the 2.21 and 14.8 ng/mL quality control (QC) samples, while the RSDs were less than 15% at the 0.554 ng/mL QC level.

  1. Determination of selected bisphenols, parabens and estrogens in human plasma using LC-MS/MS

    Czech Academy of Sciences Publication Activity Database

    Kolatorova Sosvorova, L.; Chlupacova, T.; Vitku, J.; Vlk, Martin; Heráček, J.; Stárka, L.; Šaman, David; Šimková, M.; Hampl, R.

    2017-01-01

    Roč. 174, NOV 1 (2017), s. 21-28 ISSN 0039-9140 Institutional support: RVO:61389030 ; RVO:61388963 Keywords : Alternative bisphenol * Bisphenol A * Bisphenol F * Endocrine disruptor * lc-ms/ms * Paraben Subject RIV: CC - Organic Chemistry OBOR OECD: Analytical chemistry; Organic chemistry (UOCHB-X) Impact factor: 4.162, year: 2016

  2. GC-MS-MS analysis of bacterial fatty acids in heavily creosote-contaminated soil samples

    Czech Academy of Sciences Publication Activity Database

    Byss, Marius; Tříska, Jan; Elhottová, Dana

    Č. 387 (2007), s. 1573-1577 ISSN 1618-2642 R&D Projects: GA MŠk(CZ) LC06066 Institutional research plan: CEZ:AV0Z60870520; CEZ:AV0Z60660521 Keywords : GC-MS-MS * PLFA * PAH Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 2.867, year: 2007

  3. Determination of chlormequat in pig serum and sow milk by LC-MS/MS

    DEFF Research Database (Denmark)

    Poulsen, Mette Erecius; Christensen, Hanne Bjerre; Sørensen, M.T.

    2007-01-01

    reproduction. Validation of the method was based on recovery tests at three spiking levels, determined as double determinations and repeated at least four times. Samples were extracted with methanol-water-acetic acid, centrifuged, filtrated and determined by LC-MS/MS. The mean recoveries were in the range 80...

  4. Multiple structure alignment with msTALI

    Directory of Open Access Journals (Sweden)

    Shealy Paul

    2012-05-01

    Full Text Available Abstract Background Multiple structure alignments have received increasing attention in recent years as an alternative to multiple sequence alignments. Although multiple structure alignment algorithms can potentially be applied to a number of problems, they have primarily been used for protein core identification. A method that is capable of solving a variety of problems using structure comparison is still absent. Here we introduce a program msTALI for aligning multiple protein structures. Our algorithm uses several informative features to guide its alignments: torsion angles, backbone Cα atom positions, secondary structure, residue type, surface accessibility, and properties of nearby atoms. The algorithm allows the user to weight the types of information used to generate the alignment, which expands its utility to a wide variety of problems. Results msTALI exhibits competitive results on 824 families from the Homstrad and SABmark databases when compared to Matt and Mustang. We also demonstrate success at building a database of protein cores using 341 randomly selected CATH domains and highlight the contribution of msTALI compared to the CATH classifications. Finally, we present an example applying msTALI to the problem of detecting hinges in a protein undergoing rigid-body motion. Conclusions msTALI is an effective algorithm for multiple structure alignment. In addition to its performance on standard comparison databases, it utilizes clear, informative features, allowing further customization for domain-specific applications. The C++ source code for msTALI is available for Linux on the web at http://ifestos.cse.sc.edu/mstali.

  5. Overlapping MALDI-Mass Spectrometry Imaging for In-Parallel MS and MS/MS Data Acquisition without Sacrificing Spatial Resolution

    Science.gov (United States)

    Hansen, Rebecca L.; Lee, Young Jin

    2017-09-01

    Metabolomics experiments require chemical identifications, often through MS/MS analysis. In mass spectrometry imaging (MSI), this necessitates running several serial tissue sections or using a multiplex data acquisition method. We have previously developed a multiplex MSI method to obtain MS and MS/MS data in a single experiment to acquire more chemical information in less data acquisition time. In this method, each raster step is composed of several spiral steps and each spiral step is used for a separate scan event (e.g., MS or MS/MS). One main limitation of this method is the loss of spatial resolution as the number of spiral steps increases, limiting its applicability for high-spatial resolution MSI. In this work, we demonstrate multiplex MS imaging is possible without sacrificing spatial resolution by the use of overlapping spiral steps, instead of spatially separated spiral steps as used in the previous work. Significant amounts of matrix and analytes are still left after multiple spectral acquisitions, especially with nanoparticle matrices, so that high quality MS and MS/MS data can be obtained on virtually the same tissue spot. This method was then applied to visualize metabolites and acquire their MS/MS spectra in maize leaf cross-sections at 10 μm spatial resolution. [Figure not available: see fulltext.

  6. An impulse-driven liquid-droplet deposition interface for combining LC with MALDI MS and MS/MS.

    Science.gov (United States)

    Young, J Bryce; Li, Liang

    2006-03-01

    A simple and robust impulse-driven droplet deposition system was developed for off-line liquid chromatography matrix-assisted laser desorption ionization mass spectrometry (LC-MALDI MS). The system uses a solenoid operated with a pulsed voltage power supply to generate impulses that dislodge the hanging droplets from the LC outlet directly to a MALDI plate via a momentum transfer process. There is no contact between the LC outlet and the collection surface. The system is compatible with solvents of varying polarity and viscosity, and accommodates the use of hydrophobic and hydrophilic MALDI matrices. MALDI spots are produced on-line with the separation, and do not require further processing before MS analysis. It is shown that high quality MALDI spectra from 5 fmol of pyro-Glu-fibrinopeptide deposition after LC separation could be obtained using the device, indicating that there was no sample loss in the interface. To demonstrate the analytical performance of the system as a proteome analysis tool, a range of BSA digest concentrations covering about 3 orders of magnitude, from 5 fmol to 1 pmol, were analyzed by LC-MALDI quadrupole time-of-flight MS, yielding 6 and 57% amino acid sequence coverage, respectively. In addition, a complex protein mixture of an E. coli cell extract was tryptically digested and analyzed by LC-MALDI MS, resulting in the detection of a total of 409 unique peptides from 100 fractions of 15-s intervals.

  7. An in silico MS/MS library for automatic annotation of novel FAHFA lipids.

    Science.gov (United States)

    Ma, Yan; Kind, Tobias; Vaniya, Arpana; Gennity, Ingrid; Fahrmann, Johannes F; Fiehn, Oliver

    2015-01-01

    A new lipid class named 'fatty acid esters of hydroxyl fatty acids' (FAHFA) was recently discovered in mammalian adipose tissue and in blood plasma and some FAHFAs were found to be associated with type 2 diabetes. To facilitate the automatic annotation of FAHFAs in biological specimens, a tandem mass spectra (MS/MS) library is needed. Due to the limitation of the commercial available standard compounds, we proposed building an in silico MS/MS library to extend the coverage of molecules. We developed a computer-generated library with 3267 tandem mass spectra (MS/MS) for 1089 FAHFA species. FAHFA spectra were generated based on authentic standards with negative mode electrospray ionization and 10, 20, and 40 V collision induced dissociation at 4 spectra/s as used in in ultra-high performance liquid chromatography-QTOF mass spectrometry studies. However, positional information of the hydroxyl group is only obtained either at lower QTOF spectra acquisition rates of 1 spectrum/s or at the MS(3) level in ion trap instruments. Therefore, an additional set of 4290 fragment-rich MS/MS spectra was created to enable distinguishing positional FAHFA isomers. The library was generated based on ion fragmentations and ion intensities of FAHFA external reference standards, developing a heuristic model for fragmentation rules and extending these rules to large swaths of computer-generated structures of FAHFAs with varying chain lengths, degrees of unsaturation and hydroxyl group positions. Subsequently, we validated the new in silico library by discovering several new FAHFA species in egg yolk, showing that this library enables high-throughput screening of FAHFA lipids in various biological matrices. The developed library and templates are freely available for commercial or noncommercial use at http://fiehnlab.ucdavis.edu/staff/yanma/fahfa-lipid-library. This in silico MS/MS library allows users to annotate FAHFAs from accurate mass tandem mass spectra in an easy and fast manner

  8. Determination of steroidal glycosides in Yucca gloriosa flowers by LC/MS/MS.

    Science.gov (United States)

    Montoro, Paola; Skhirtladze, Alexandre; Perrone, Angela; Benidze, Mariam; Kemertelidze, Ether; Piacente, Sonia

    2010-09-05

    An high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) method, was developed for the quantitative analysis of the steroidal glycosides occurring in Yucca gloriosa flowers. The HPLC experiments were performed by means of an octadecyl-modified reversed-phase C-18 column and a binary mobile phase system under gradient elution conditions. The fragmentation patterns of steroidal saponins were analyzed by ESI-MS(n) in positive ion mode and a specific multiple reaction monitoring MS/MS detection was developed for their quantitative determination. The described method provides high sensitivity and specificity for quantitative determination of the steroidal glycosides in Y. gloriosa flowers. Quantification was performed against an external calibration line obtained using each pure steroidal glycoside. Short- and long-term repeatabilities of the methods were better than 3 and 6%, respectively. The method was validated according to EMEA guidelines and applied to real samples. 2010 Elsevier B.V. All rights reserved.

  9. MS/MS similarity networking accelerated target profiling of triterpene saponins in Eleutherococcus senticosus leaves.

    Science.gov (United States)

    Ge, Yue-Wei; Zhu, Shu; Yoshimatsu, Kayo; Komatsu, Katsuko

    2017-07-15

    The targeted mass information of compounds accelerated their discovery in a large volume of untargeted MS data. An MS/MS similarity networking is advanced in clustering the structural analogues, which benefits the collection of mass information of similar compounds. The triterpene saponins extracted from Eleutherococcus senticosus leaves (ESL), a kind of functional tea, have shown promise in the relief of Alzheimer's disease. In this work, a target-precursor list (TPL) generated using MS/MS similarity networking was employed to rapidly trace 106 triterpene saponins from the aqueous extracts of ESL, of which 49 were tentatively identified as potentially new triterpene saponins. Moreover, a compound database of triterpene saponins was established and successfully applied to uncover their distribution features in ESL samples collected from different areas. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Dopamine, T cells and multiple sclerosis (MS).

    Science.gov (United States)

    Levite, Mia; Marino, Franca; Cosentino, Marco

    2017-05-01

    Dopamine is a key neurotransmitter that induces critical effects in the nervous system and in many peripheral organs, via 5 dopamine receptors (DRs): D1R-D5R. Dopamine also induces many direct and very potent effects on many DR-expressing immune cells, primarily T cells and dendritic cells. In this review, we focus only on dopamine receptors, effects and production in T cells. Dopamine by itself (at an optimal concentration of~0.1 nM) induces multiple function of resting normal human T cells, among them: T cell adhesion, chemotactic migration, homing, cytokine secretion and others. Interestingly, dopamine activates resting effector T cells (Teffs), but suppresses regulatory T cells (Tregs), and both effects lead eventually to Teff activation. Dopamine-induced effects on T cells are dynamic, context-sensitive and determined by the: T cell activation state, T cell type, DR type, and dopamine concentration. Dopamine itself, and also few dopaminergic molecules/ drugs that are in clinical use for cardiac, neurological and other non-immune indications, have direct effects on human T cells (summarized in this review). These dopaminergic drugs include: dopamine = intropin, L-DOPA, bromocriptine, pramipexole, pergolide, haloperidol, pimozide, and amantadine. Other dopaminergic drugs were not yet tested for their direct effects on T cells. Extensive evidence in multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE) show dopaminergic dysregulations in T cells in these diseases: D1-like DRs are decreased in Teffs of MS patients, and dopamine does not affect these cells. In contrast, D1-like DRs are increased in Tregs of MS patients, possibly causing functional Treg impairment in MS. Treatment of MS patients with interferon β (IFN-β) increases D1-like DRs and decreases D2-like DRs in Teffs, decreases D1-like DRs in Tregs, and most important: restores responsiveness of patient's Teffs to dopamine. DR agonists and antagonists confer some benefits in

  11. Highly sensitive and specific analysis of sterol profiles in biological samples by HPLC-ESI-MS/MS.

    Science.gov (United States)

    Honda, Akira; Miyazaki, Teruo; Ikegami, Tadashi; Iwamoto, Junichi; Yamashita, Kouwa; Numazawa, Mitsuteru; Matsuzaki, Yasushi

    2010-08-01

    High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) is a powerful method for the microanalysis of compounds in biological samples. Compared with gas chromatography-mass spectrometry (GC-MS), this method is more broadly applicable to various compounds and usually does not require a derivatization step before analysis. However, when neutral sterols are analyzed, the sensitivities of usual HPLC-MS/MS method are not superior to those of GC-MS because the sterols are relatively resistant to ionization. In this review, we introduce the recent development of HPLC-MS/MS analysis for the quantification of non-cholesterol sterols. By adding an effective derivatization step to the conventional procedure, sterol analysis by HPLC-MS/MS surpassed that obtained by GC-MS in sensitivity. In addition, sufficient specificity of this method was achieved by selected reaction monitoring (SRM) and thorough chromatographic separation of each sterol. Copyright 2010 Elsevier Ltd. All rights reserved.

  12. Metabolic profile and pharmacokinetics of polyphyllin I, an anticancer candidate, in rats by UPLC-QTOF-MS/MS and LC-TQ-MS/MS.

    Science.gov (United States)

    Liu, Yu-Cai; Zhu, He; Shakya, Shailendra; Wu, Jun-Wen

    2017-03-01

    Polyphyllin I (PPI), a natural steroidal saponin originating from rihzome of Paris polyphylla, is a potential anticancer candidate. Previous pharmacokinetics study showed that the oral bioavailability of PPI was very low, which suggested that certain amount of PPI might be metabolized in vivo. However, to date, information regarding the final metabolic fates of PPI is very limited. In this study, metabolites of PPI and their pharmacokinetics in rats were investigated using UPLC-QTOF-MS/MS and LC-TQ-MS/MS. A total of seven putative metabolites, including six phase I and one phase II metabolites, were detected and identified with three exact structures by comparison with authentic standards for the first time. Oxidation, deglycosylation and glucuronidation were found to be the major metabolic processes of the compound in rats. The pharmacokinetics of prosapogenin A, trillin and diosgenin, three deglycosylation metabolites of PPI with definite anticancer effects, were further studied, which suggested that the metabolites underwent a prolonged absorption and slower elimination after intragastric administration of PPI at the dose of 500 mg/kg. This study provides valuable and new information on the metabolic fate of PPI, which will be helpful in further understanding its mechanism of action. Copyright © 2016 John Wiley & Sons, Ltd.

  13. 2014 USGS/NRCS Lidar: Central MS

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — TASK NAME: USGS-NRCS Laurel MS 0.7m NPS LIDAR Lidar Data Acquisition and Processing Production Task USGS Contract No. G10PC00057 Task Order No. G13PD01086 Woolpert...

  14. Interpretation of MS ISO/ IEC 17020

    International Nuclear Information System (INIS)

    Mohamad Pauzi Ismail

    2012-01-01

    MS ISO/ IEC 17020 is an international standard which adopted by Malaysian Standard states the requirement criteria for inspection bodies. The author involved in the NDT technical assessment to several NDT companies in Malaysia. The experience in performing NDT assessment activity is explained and discussed. (author)

  15. Retinoid quantification by HPLC/MS(n)

    Science.gov (United States)

    McCaffery, Peter; Evans, James; Koul, Omanand; Volpert, Amy; Reid, Kevin; Ullman, M. David

    2002-01-01

    Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.

  16. Antibody-free LC-MS/MS quantification of rhTRAIL in human and mouse serum

    NARCIS (Netherlands)

    Wilffert, Daniel; Reis, C.R.; Hermans, Jos; Govorukhina, Natalia; Tomar, Tushar; de Jong, Steven; Quax, Wim J.; van de Merbel, Nico C.; Bischoff, Rainer

    2013-01-01

    The major challenge in targeted protein quantification by LC-MS/MS in serum lies in the complexity of the biological matrix with regard to the wide diversity of proteins and their extremely large dynamic concentration range. In this study, an LC-MS/MS method was developed for the simultaneous

  17. Factors Associated with Neurologists' Provision of MS Patient Care

    Science.gov (United States)

    Halpern, Michael T.; Teixeira-Poit, Stephanie M.; Kane, Heather; Frost, Corey; Keating, Michael; Olmsted, Murrey

    2014-01-01

    Neurologists are central to providing quality care for individuals with MS. However, neurologist shortages may restrict access to care for MS patients. To examine factors influencing neurologists' provision of MS care, we surveyed 1,700 US neurologists to assess demographic/practice characteristics, training, and attitudes toward MS care. The study population consisted of 573 respondents: 87 (15.2%) MS subspecialists and 486 (84.8%) “other neurologists,” including subspecialists in other neurology areas (i.e., non-MS) and general neurologists. MS subspecialists indicating they “enjoy interacting with MS patients” had a significantly greater rate of MS patients seen per week. In separate analyses of the “other neurologists” group, the rate of MS patients seen was lower among neurologists in university-based groups or those practicing in major cities; female neurologists; and neurologists who indicated lack of sufficient knowledge regarding MS patient care. Rates of MS patients seen were significantly greater for other neurologists who agreed that MS care involved “ability to improve patient outcomes and quality of life”; “dynamic area with evolving treatment options”; and “enjoy interacting with MS patients.” Understanding factors influencing MS patient care by neurologists and developing policies for appropriate access to care is critical for optimal outcomes among this population. PMID:24949203

  18. Factors Associated with Neurologists’ Provision of MS Patient Care

    Directory of Open Access Journals (Sweden)

    Michael T. Halpern

    2014-01-01

    Full Text Available Neurologists are central to providing quality care for individuals with MS. However, neurologist shortages may restrict access to care for MS patients. To examine factors influencing neurologists’ provision of MS care, we surveyed 1,700 US neurologists to assess demographic/practice characteristics, training, and attitudes toward MS care. The study population consisted of 573 respondents: 87 (15.2% MS subspecialists and 486 (84.8% “other neurologists,” including subspecialists in other neurology areas (i.e., non-MS and general neurologists. MS subspecialists indicating they “enjoy interacting with MS patients” had a significantly greater rate of MS patients seen per week. In separate analyses of the “other neurologists” group, the rate of MS patients seen was lower among neurologists in university-based groups or those practicing in major cities; female neurologists; and neurologists who indicated lack of sufficient knowledge regarding MS patient care. Rates of MS patients seen were significantly greater for other neurologists who agreed that MS care involved “ability to improve patient outcomes and quality of life”; “dynamic area with evolving treatment options”; and “enjoy interacting with MS patients.” Understanding factors influencing MS patient care by neurologists and developing policies for appropriate access to care is critical for optimal outcomes among this population.

  19. EXTRACTION AND DETECTION OF A NEW ARSINE SULFIDE CONTAINING ARSENOSUGAR IN MOLLUSCS BY IC-ICP-MS AND IC-ESI-MS/MS

    Science.gov (United States)

    Using IC-ICP-MS and IC-ESI-MS/MS, an unknown arsenical compound in mollusks has been identified as a new arsine sulfide containing analog of a known arsenosugar and is referred to as As(498). This species has been observed in four separate shellfish species following a mild metha...

  20. Quantitative Determination of Perfluorochemicals and Fluorotelomer Alcohols in Plants from Biosolid-Amended Fields using LC/MS/MS and GC/MS

    Science.gov (United States)

    Analytical methods for determining perfluorochemicals (PFCs) and fluorotelomer alcohols (FTOHs) in plants using liquid chromatography/tandem mass spectrometry (LC/MS/MS) and gas chromatography/mass spectrometry (GC/MS) were developed, and applied to quantify a suite of analytes i...

  1. MS+MS/MS

    Indian Academy of Sciences (India)

    2014-04-12

    ,6. 0. 12. 100. 72. 100 subunit, mitochondrial OS=Mus musculus GN=Sdha PE. Peptide Information. Calc. Mass. Obsrv. Mass. ± da ± ppm. Start. End Sequence. Ion. C. I. %. Modification. Rank Result Type. Seq. Seq. Score.

  2. MS+MS/MS

    Indian Academy of Sciences (India)

    2014-04-12

    /Pos. 130/F5. Instr./Gel Origin. AK139/T056. Process Status Analysis Succeeded. Plate [#] Name. [1] T056P10. Instrument Sample Name. Spectra. 9. Rank. Protein Name. Accession No. Protein. Protein MW Protein. Pep.

  3. Rapid Determination of Isomeric Benzoylpaeoniflorin and Benzoylalbiflorin in Rat Plasma by LC-MS/MS Method

    Directory of Open Access Journals (Sweden)

    Chuanqi Zhou

    2017-01-01

    Full Text Available Benzoylpaeoniflorin (BP is a potential therapeutic agent against oxidative stress related Alzheimer’s disease. In this study, a more rapid, selective, and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS method was developed to determine BP in rat plasma distinguishing with a monoterpene isomer, benzoylalbiflorin (BA. The method showed a linear response from 1 to 1000 ng/mL (r>0.9950. The precision of the interday and intraday ranged from 2.03 to 12.48% and the accuracy values ranged from −8.00 to 10.33%. Each running of the method could be finished in 4 minutes. The LC-MS/MS method was validated for specificity, linearity, precision, accuracy, recovery, and stability and was found to be acceptable for bioanalytical application. Finally, this fully validated method was successfully applied to a pharmacokinetic study in rats following oral administration.

  4. Determination of drug residues by CLAR-MS/MS in animal tissues

    International Nuclear Information System (INIS)

    Brenes Jimenez, Jose Eduardo

    2009-01-01

    Produced food of animal origin, present the possibility of occurrence of any contact with substances that have negative effects on the health of people who consume them. The use of drugs in veterinary medicine is one of the possible sources of such waste; so, the conditions for the analysis of some classes of antibiotics in animal tissues are based on the study. Costa Rica and the countries that are export destination, have regulation and programs for control before to be distributed in local markets, or post if it is received any complaint of pollution. The high resolution liquid chromatography coupled to mass spectrometers (CLAR-MS/MS) allows the analysis of analytes monitored, according to the specifications required by the legislation. The cases of two laboratories in Costa Rica are presented as the only ones who have the ability to perform the analysis of drug residues CLAR-MS/MS. (author) [es

  5. Mannose7 Glycan Isomer Characterization by IMS-MS/MS Analysis

    Science.gov (United States)

    Zhu, Feifei; Lee, Sunyoung; Valentine, Stephen J.; Reilly, James P.; Clemmer, David E.

    2012-12-01

    The isomers of the Man7GlcNAc2 glycan obtained from bovine ribonuclease B have been characterized by ion mobility spectrometry-tandem mass spectrometry (IMS-MS/MS). In these experiments, [Man7 + 2Na]2+ precursors having different mobilities are selected by ion mobility spectrometry and analyzed by MS/MS techniques in an ion trap. The fragmentation spectra obtained for various precursor ions are specific, suggesting the isolation or enrichment of different glycan isomers. One fragment ion with a mass-to-charge ratio ( m/z) of 903.8 is found to correspond to the loss of an internal mannose residue of a specific isomer. Extracted fragment ion drift time distributions (XFIDTDs) yield distinctive precursor ion drift time profiles indicating the existence of four separate isomers as proposed previously.

  6. A Rapid and Sensitive LC-MS/MS Method for the Determination of ...

    African Journals Online (AJOL)

    VIS) detection of the analytes is also reported. Sulfonamides exhibited higher response factors towards UV/VIS than mass spectrometer detection and the opposite was true for the rest of the analytes. KEYWORDS: Quadrupole ion trap, LC-MS, ...

  7. How much separation for LC-MS/MS quantitative bioanalysis of drugs and metabolites?

    Science.gov (United States)

    Tan, Aimin; Fanaras, John C

    2018-05-01

    LC-MS/MS has been the dominant analytical technology for quantitative bioanalysis of drugs and metabolites for more than two decades. Despite this, a very fundamental question like how much separation is required for LC-MS/MS quantitative bioanalysis of drugs and metabolites has not been adequately addressed. Some think that no or only very limited separation is necessary thanks to the unparalleled selectivity offered by tandem mass spectrometry. Others think that the more separation, the better, because of the potential detrimental impact of matrix effect (ion suppression or enhancement). Still others just use a rule-of-thumb approach by keeping the adjusted retention/capacity factor always between 2 and 5. The purpose of this article is to address this fundamental question through rational thinking together with various real case examples drawn from regulated bioanalytical laboratories. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. [Analysis of chemical constituents from Citrus aurantium by UHPLC-LTQ-Orbitrap-MS/MS].

    Science.gov (United States)

    Yu, Guo-Hua; Yang, Hong-Jun; Li, Jun-Fang; Geng, Ya; Dong, Ling; Xu, Hai-Yu

    2016-09-01

    In this study, a ultra-high-pressure liquid chromatography coupled with linear ion trap-Orbitrap tandem mass spectrometry (UHPLC-LTQ-Orbitrap-MS/MS) method was developed to analyze the chemical components in Citrus aurantium. C. aurantium were extracted with 75% methanol, and we applied a Thermo Scientific BDS Hypersil C₁₈ column (2.1 mm×150 mm, 2.4 μm) to UHPLC analysis with acetonitrile-water (containing 0.1% formic acid) in gradient as mobile phase. Elutes were then detected by ESI-LTQ-Orbitrap-MS/MS. A total of 27 components were identified, including fourteen flavonoids, seven coumarins, five limonoids and one alkaloid. This study showed an insight into the composition of C. aurantium, which could provide theoretical foundation for further study and utilization of the medicinal resources. Copyright© by the Chinese Pharmaceutical Association.

  9. Analytical Method Details (MS): SE51_MS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available MS ESI N.A. The metabolites were analyzed using a liquid chromatography quadrupole time of flight mass spectrometer... and tandem quadruple mass spectrometer system (UPLC-QTOF Premier and UPLC-TQS, Waters Co., MA, USA)

  10. QuaMeter: multivendor performance metrics for LC-MS/MS proteomics instrumentation.

    Science.gov (United States)

    Ma, Ze-Qiang; Polzin, Kenneth O; Dasari, Surendra; Chambers, Matthew C; Schilling, Birgit; Gibson, Bradford W; Tran, Bao Q; Vega-Montoto, Lorenzo; Liebler, Daniel C; Tabb, David L

    2012-07-17

    LC-MS/MS-based proteomics studies rely on stable analytical system performance that can be evaluated by objective criteria. The National Institute of Standards and Technology (NIST) introduced the MSQC software to compute diverse metrics from experimental LC-MS/MS data, enabling quality analysis and quality control (QA/QC) of proteomics instrumentation. In practice, however, several attributes of the MSQC software prevent its use for routine instrument monitoring. Here, we present QuaMeter, an open-source tool that improves MSQC in several aspects. QuaMeter can directly read raw data from instruments manufactured by different vendors. The software can work with a wide variety of peptide identification software for improved reliability and flexibility. Finally, QC metrics implemented in QuaMeter are rigorously defined and tested. The source code and binary versions of QuaMeter are available under Apache 2.0 License at http://fenchurch.mc.vanderbilt.edu.

  11. Analytical Method Details (MS): SE59_MS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available argeted metabolomic analysis using LC-QTOF-MS and the metabolome data processing of alignment, de-isotoping, noise cutting, and normalization was conducted as previously described. ...

  12. A survey of free glutamic acid in foods using a robust LC-MS/MS method.

    Science.gov (United States)

    Cebi, Nur; Dogan, Canan Ekinci; Olgun, Elmas Oktem; Sagdic, Osman

    2018-05-15

    An effective and simultaneous liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used with the aim of quantifying monosodium glutamate (MSG) in foodstuffs, such as chips, taste cubes, sauces and soups. The results were linear (R 2  = 1), with very low LOD and LOQ values, 1.0 µg/kg, 5.0 µg/kg, respectively. Excellent repeatability and reproducibility were also achieved. This highly sensitive and robust LC-MS/MS technique was applied successfully for the detection and quantification of MSG in a wide variety of foodstuffs. MSG contents ranged from 0.01 g/100 g to 15.39 g/100 g in food samples. Importantly, determination of free glutamic acid in the daily diet could also prevent various side effects associated with consumption of excess free glutamic acid. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. The APEX Quantitative Proteomics Tool: Generating protein quantitation estimates from LC-MS/MS proteomics results

    Directory of Open Access Journals (Sweden)

    Saeed Alexander I

    2008-12-01

    Full Text Available Abstract Background Mass spectrometry (MS based label-free protein quantitation has mainly focused on analysis of ion peak heights and peptide spectral counts. Most analyses of tandem mass spectrometry (MS/MS data begin with an enzymatic digestion of a complex protein mixture to generate smaller peptides that can be separated and identified by an MS/MS instrument. Peptide spectral counting techniques attempt to quantify protein abundance by counting the number of detected tryptic peptides and their corresponding MS spectra. However, spectral counting is confounded by the fact that peptide physicochemical properties severely affect MS detection resulting in each peptide having a different detection probability. Lu et al. (2007 described a modified spectral counting technique, Absolute Protein Expression (APEX, which improves on basic spectral counting methods by including a correction factor for each protein (called Oi value that accounts for variable peptide detection by MS techniques. The technique uses machine learning classification to derive peptide detection probabilities that are used to predict the number of tryptic peptides expected to be detected for one molecule of a particular protein (Oi. This predicted spectral count is compared to the protein's observed MS total spectral count during APEX computation of protein abundances. Results The APEX Quantitative Proteomics Tool, introduced here, is a free open source Java application that supports the APEX protein quantitation technique. The APEX tool uses data from standard tandem mass spectrometry proteomics experiments and provides computational support for APEX protein abundance quantitation through a set of graphical user interfaces that partition thparameter controls for the various processing tasks. The tool also provides a Z-score analysis for identification of significant differential protein expression, a utility to assess APEX classifier performance via cross validation, and a

  14. Analytical Method Details (MS): SE8_MS4 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE8_MS4 LC-FT-ICR-MS ESI positive method 4 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...ment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100Z (4.6 x ...e solved in 300uL 80% methanol solution. 20uL sample is injected into HPLC after 0.2um membrane filter treat

  15. Analytical Method Details (MS): SE8_MS3 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE8_MS3 LC-FT-ICR-MS ESI positive method 3 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...ment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100Z (4.6 x ...e solved in 300uL 80% methanol solution. 20uL sample is injected into HPLC after 0.2um membrane filter treat

  16. Analytical Method Details (MS): SE8_MS2 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE8_MS2 LC-FT-ICR-MS ESI positive method 2 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...ment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100Z (4.6 x ...e solved in 300uL 80% methanol solution. 20uL sample is injected into HPLC after 0.2um membrane filter treat

  17. Analytical Method Details (MS): SE4_MS3 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE4_MS3 LC-FT-ICR-MS ESI positive method 3 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...olved in 300uL 80% methanol solution. 20uL sample is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agile...nt 1100 series (Agilent), Column: TSKgel-100V (4.6 x 250 mm, 5 micrometer; TOSOH),

  18. Analytical Method Details (MS): SE8_MS1 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available SE8_MS1 LC-FT-ICR-MS ESI positive method 1 Agilent1100 HPLC (Agilent), LTQ-FT (Ther...ment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100Z (4.6 x ...e solved in 300uL 80% methanol solution. 20uL sample is injected into HPLC after 0.2um membrane filter treat

  19. Evaluation of Tamoxifen and metabolites by LC-MS/MS and HPLC Methods

    OpenAIRE

    Heath, D.D.; Flatt, S.W.; Wu, A.H.B.; Pruitt, M.A.; Rock, C.L.

    2014-01-01

    Epidemiological and laboratory evidence suggests that quantification of serum or plasma levels of tamoxifen and the metabolites of tamoxifen, 4-hydroxy-N-desmethyl-tamoxifen (endoxifen), Z-4-hydroxy-tamoxifen (4HT), N-desmethyl-tamoxifen (ND-tam) is a clinically useful tool in the assessment and monitoring of breast cancer status in patients taking adjuvant tamoxifen. A liquid chromatographic mass spectrometric method (LC-MS/MS) was used to measure the blood levels of tamoxifen and the metabo...

  20. STS-33 MS Carter and MS Thornton display 'Maggot on Board' sign and candy

    Science.gov (United States)

    1989-01-01

    STS-33 Mission Specialist (MS) Manley L. Carter, Jr (left) and MS Kathryn C. Thornton display 'Maggot on Board' sign and 'SMARTIES' candy stored in plastic bag on the aft flight deck of Discovery, Orbiter Vehicle (OV) 103. The mission specialists are wearing their mission polo shirts and communications kit assembly headsets. An overhead window appears above their heads. A gold necklace chain floats around Carter's neck.

  1. Identification and Quantification of Several Mammalian Steroid Hormones in Plants by UPLC-MS/MS

    Czech Academy of Sciences Publication Activity Database

    Simerský, Radim; Novák, Ondřej; Morris, David; Pouzar, Vladimír; Strnad, Miroslav

    2009-01-01

    Roč. 28, č. 2 (2009), s. 125-136 ISSN 0721-7595 R&D Projects: GA AV ČR KAN200380801 Institutional research plan: CEZ:AV0Z50380511; CEZ:AV0Z40550506 Keywords : Ultra-performance liquid chromatography (UPLC) * Tandem mass spectrometry (MS/MS) * Immunoaffinity purification * Steroids * Plant extracts * Digitalis purpurea * Nicotiana tabacum * Inula helenium Subject RIV: EC - Immunology Impact factor: 2.438, year: 2009

  2. Quantifying MMA by SLE LC-MS/MS: Unexpected challenges in assay development.

    Science.gov (United States)

    Lo, Sheng-Ying; Gordon, Cindy; Sadilkova, Katerina; Jack, Rhona M; Dickerson, Jane A

    2016-09-01

    Analysis of serum/plasma methylmalonic acid (MMA) is important for the diagnosis and management of methylmalonic acidemia in pediatric populations. This work focuses on developing and validating a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to monitor methylmalonic acidemia using a simple method preparation. MMA and stable isotope labeled d3-MMA was extracted using supported liquid extraction (SLE). Assay imprecision, bias, linearity, recovery and carryover were determined. The relationship between MMA and propionyl acylcarnitine (C3-acylcarnitine) was also evaluated using historical paired results from 51 unique individuals. Baseline separation between MMA and succinic acid was completed in 7min. The assay was linear from 0.1 to 500μM. The intra-day and inter-day imprecision CV ranged from 4.1 to 13.2% (0.3 to 526μM) and 5.0 to 15.7% (0.3 to 233μM), respectively. Recovery ranged from 93 to 125%. The correlation with a national reference laboratory LC-MS/MS assay showed a Deming regression of 1.026 and intercept of -1.335. Carryover was determined to be MMA and C3-acylcarnitine. This report describes the first LC-MS/MS method using SLE for MMA extraction. In addition, we illustrate the challenges encountered during this method development that should be assessed and resolved by any laboratory implementing a SLE LC-MS/MS assay designed to quantify analytes across several orders of magnitude. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  3. New method for quantification of gasotransmitter hydrogen sulfide in biological matrices by LC-MS/MS

    OpenAIRE

    Bo Tan; Sheng Jin; Jiping Sun; Zhongkai Gu; Xiaotian Sun; Yichun Zhu; Keke Huo; Zonglian Cao; Ping Yang; Xiaoming Xin; Xinhua Liu; Lilong Pan; Furong Qiu; Jian Jiang; Yiqun Jia

    2017-01-01

    Hydrogen sulfide exists widely in mammalian tissues and plays a vital role in physiological and pathophysiological processes. However, striking differences with orders of magnitude were observed for the detected hydrogen sulfide concentrations in biological matrices among different measurements in literature, which lead to the uncertainty for examination the biological relevance of hydrogen sulfide. Here, we developed and validated a liquid chromatography- mass spectrometry (LC-MS/MS) method ...

  4. Analytical Method Details (MS): SE58_MS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available Data acquisition was performed on a Pegasus IV TOF mass spectrometer (LECO, St. Joseph, MI, USA) with an acq...SE58_MS01 GC-TOF-MS Agilent 6890N gas chromatograph (Agilent Technologies) and Pegasus IV TOF mass spectrome...ter (LECO) EI Positive For metabolome analysis, 1 μL of extracts (∼5.6 μg of each s

  5. Relationship between plasma and salivary melatonin and cortisol investigated by LC-MS/MS.

    Science.gov (United States)

    van Faassen, Martijn; Bischoff, Rainer; Kema, Ido P

    2017-08-28

    Disturbance of the circadian rhythm has been associated with disease states, such as metabolic disorders, depression and cancer. Quantification of the circadian markers such as melatonin and cortisol critically depend on reliable and reproducible analytical methods. Previously, melatonin and cortisol were primarily analyzed separately, mainly using immunoassays. Here we describe the validation and application of a high-throughput liquid chromatography in combination with mass spectrometry (LC-MS/MS) method for the combined analysis of melatonin and cortisol in plasma and saliva. The LC-MS/MS method was validated according to international validation guidelines. We used this method to analyze total plasma, free plasma (as obtained by equilibrium dialysis) and saliva melatonin and cortisol in healthy adults. Validation results for plasma and saliva melatonin and cortisol were well within the international validation criteria. We observed no difference between saliva collected by passive drooling or Salivette. Moreover, we noted a significant difference in saliva vs. free plasma melatonin. We observed on average 36% (95% CI: 4%-60%) higher salivary melatonin levels in comparison to free plasma melatonin, suggestive of local production of melatonin in the salivary glands. The novel outcome of this study is probably due to the high precision of our LC-MS/MS assay. These outcomes illustrate the added value of accurate and sensitive mass spectrometry based methods for the quantification of neuroendocrine biomarkers.

  6. Quantitative LC-MS/MS Analysis of Proteins Involved in Metastasis of Breast Cancer

    Science.gov (United States)

    Goto, Rieko; Nakamura, Yasushi; Takami, Tomonori; Sanke, Tokio; Tozuka, Zenzaburo

    2015-01-01

    The purpose of this study was to develop quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for the analysis of proteins involved in metastasis of breast cancer for diagnosis and determining disease prognosis, as well as to further our understand of metastatic mechanisms. We have previously demonstrated that the protein type XIV collagen may be specifically expressed in metastatic tissues by two dimensional LC-MS/MS. In this study, we developed quantitative LC-MS/MS methods for type XIV collagen. Type XIV collagen was quantified by analyzing 2 peptides generated by digesting type XIV collagen using stable isotope-labeled peptides. The individual concentrations were equivalent between 2 different peptides of type XIV collagen by evaluation of imprecise transitions and using the best transition for the peptide concentration. The results indicated that type XIV collagen is highly expressed in metastatic tissues of patients with massive lymph node involvement compared to non-metastatic tissues. These findings were validated by quantitative real-time RT-PCR. Further studies on type XIV collagen are desired to verify its role as a prognostic factor and diagnosis marker for metastasis. PMID:26176947

  7. The study of MS patients’ life style referred to MS Association [Tehran City, 2008

    Directory of Open Access Journals (Sweden)

    Firuzeh Payamani

    2016-01-01

    Full Text Available Multiple sclerosis [M.S] is one of the common autoimmune diseases that effects on central nervous system. It is unpredictable and cause to changes in life style. M.S progresses to inability. There is a close relation between health and life style, so that we can prevent from rate of disease attacks. Therefore recognition life style in MS patients is very important. The objective of this study was to determine of MS patient's life style referred to MS Association in Tehran City in 2008. This research was a descriptive and analytical study. In this study2oo men and women who were suffering from M.S disease were selected. Study instrument was questionnaire and method was interviews. Data were obtained through easy; the data were analyzed with descriptive statistical and inferential statistical methods. Findings showed there were significant relation between educational level, supportive resources, number of relapse and self-care dimension in life style. Also, relation between marriage status, family income and nutrition dimension in life style was significant. According to the results of this study, life style in some dimensions [e.g. Physical activity, exercise. Sleep and rest patterns and adaptation with stress] were undesirable. These no healthy behaviors can effect on severity and relapse of disease and ultimately these effect on quality of life in these patients. Therefore, change and modification and improvement in life style in these patients seem to be essential that we can access to it with holistic educational programs.

  8. An improved method of sample preparation on AnchorChip targets for MALDI-MS and MS/MS and its application in the liver proteome project

    DEFF Research Database (Denmark)

    Zhang, Xumin; Shi, Liang; Shu, Shaokung

    2007-01-01

    An improved method for sample preparation for MALDI-MS and MS/MS using AnchorChip targets is presented. The method, termed the SMW method (sample, matrix wash), results in better sensitivity for peptide mass fingerprinting as well as for sequencing by MS/MS than previously published methods....... The method allows up-concentration and desalting directly on the mass spectrometric target and should be amenable for automation. A draw back caused by extensive oxidation of methionine and tryptophan in the SMW method can be alleviated by the addition of n-octyl glucopyranoside and DTT to the sample...

  9. ESI-MS, ESI-MS/MS fingerprint and LC-ESI-MS analysis of proathocyanidins from Bursera simaruba Sarg bark.

    Science.gov (United States)

    Maldini, Mariateresa; Montoro, Paola; Piacente, Sonia; Pizza, Cosimo

    2009-12-01

    Direct flow injection/electrospray ionization/ion trap tandem mass spectrometry was used to investigate the presence of proanthocyanidins (PAs) in the methanolic extract of B. simaruba bark. Additionally, an LC-ESI-MS qualitative study was performed by using a monolithic stationary phase. The fragmentation pattern obtained evidenced the presence in B. simaruba bark of PAs belonging to the series of polymers of epicatechin, along with their glycosilated derivatives.

  10. Economic costs associated with an MS relapse

    LENUS (Irish Health Repository)

    O'Connell, K.

    2014-09-01

    This was an prospective audit composed of medical chart review and patient questionnaire. Relapses were stratified into 3 groups: low, moderate and high intensity. Age, gender, MS subtype, disease duration, expanded disability status scale (EDSS) score, disease modifying therapy (DMT) use and employment status were recorded. Direct costs included GP visits, investigations, clinic visit, consultations with medical staff, medication and admission costs. Indirect costs assessed loss of earnings, partner\\'s loss of earnings, childcare, meals and travel costs.

  11. Familiar Face Detection in 180ms

    Science.gov (United States)

    Visconti di Oleggio Castello, Matteo; Gobbini, M. Ida

    2015-01-01

    The visual system is tuned for rapid detection of faces, with the fastest choice saccade to a face at 100ms. Familiar faces have a more robust representation than do unfamiliar faces, and are detected faster in the absence of awareness and with reduced attentional resources. Faces of family and close friends become familiar over a protracted period involving learning the unique visual appearance, including a view-invariant representation, as well as person knowledge. We investigated the effect of personal familiarity on the earliest stages of face processing by using a saccadic-choice task to measure how fast familiar face detection can happen. Subjects made correct and reliable saccades to familiar faces when unfamiliar faces were distractors at 180ms—very rapid saccades that are 30 to 70ms earlier than the earliest evoked potential modulated by familiarity. By contrast, accuracy of saccades to unfamiliar faces with familiar faces as distractors did not exceed chance. Saccades to faces with object distractors were even faster (110 to 120 ms) and equivalent for familiar and unfamiliar faces, indicating that familiarity does not affect ultra-rapid saccades. We propose that detectors of diagnostic facial features for familiar faces develop in visual cortices through learning and allow rapid detection that precedes explicit recognition of identity. PMID:26305788

  12. Hearing Disorders in Multiple Sclerosis (MS Patiants

    Directory of Open Access Journals (Sweden)

    Ghasem Mohammadkhani

    2003-12-01

    Full Text Available Objective: Multiple sclerosis (MS is a neurological disease that causes sudden deaf. In this research auditory disorders were studied in the patients with multiple sclerosis (MS. Materials & Methods: This cross sectional descriptive-analytical study was performed on 107 patients with multiple sclerosis in the range of 20-45 yrs. There were not history of trauma and middle ear disease in all cases. Sampling was randomized. A complete auditory evaluation including pure tone audiometry, speech audiometry, imittance audiometry and brainstem Auditory Evoked Potential were performed on patients. Results: In pure tone audiometry 62.19% of cases had sensory neural high tone loss. In speech audiometry and imittance audiometry 18 and 31 cases were abnormal, respectively. 14.55% of cases had abnormality in BAEP. The most of abnormalities were prolonged latency of V, decrease of V/I amplitude ratio and poor reproducibility, respectively. With high rate of stimulation 57.77% of cases were abnormal. Statistical analysis showed significant difference between latency of V and stimulation rate. Conclusion: According to findings of this research it seems that hearing evaluation is very important for follow-up and early rehabilitation of auditory disorder in patients with MS. Also auditory tests battery especially brain-stem auditory evoked potentials (BAEP with high stimulation rate are useful in the diagnosis of Multiple Sclerosis.

  13. Pattern II and pattern III MS are entities distinct from pattern I MS: evidence from cerebrospinal fluid analysis.

    Science.gov (United States)

    Jarius, S; König, F B; Metz, I; Ruprecht, K; Paul, F; Brück, W; Wildemann, B

    2017-08-29

    The diagnosis of multiple sclerosis (MS) is currently based solely on clinical and magnetic resonance imaging features. However, histopathological studies have revealed four different patterns of lesion pathology in patients diagnosed with MS, suggesting that MS may be a pathologically heterogeneous syndrome rather than a single disease entity. The aim of this study was to investigate whether patients with pattern I MS differ from patients with pattern II or III MS with regard to cerebrospinal fluid (CSF) findings, especially with reference to intrathecal IgG synthesis, which is found in most patients with MS but is frequently missing in MS mimics such as aquaporin-4-IgG-positive neuromyelitis optica spectrum disorders and myelin oligodendrocyte glycoprotein-IgG-positive encephalomyelitis. Findings from 68 lumbar punctures in patients who underwent brain biopsy as part of their diagnostic work-up and who could be unequivocally classified as having pattern I, pattern II or pattern III MS were analysed retrospectively. Oligoclonal bands (OCBs) were present in 88.2% of samples from pattern I MS patients but in only 27% of samples from patients with pattern II or pattern III MS (P pattern II or III MS patients tested (P pattern II and III MS patients (P pattern II and III MS samples than in pattern I MS samples (P pattern II or pattern III MS differ significantly from patients with pattern I MS as well as from previous, histologically non-classified MS cohorts with regard to both intrathecal IgG synthesis and blood-CSF barrier function. Our findings strongly corroborate the notion that pattern II and pattern III MS are entities distinct from pattern I MS.

  14. Development and Validation of an LC-MS/MS Method and Comparison with a GC-MS Method to Measure Phenytoin in Human Brain Dialysate, Blood, and Saliva

    Directory of Open Access Journals (Sweden)

    Raphael Hösli

    2018-01-01

    Full Text Available Phenytoin (PHT is one of the most often used critical dose drugs, where insufficient or excessive dosing can have severe consequences such as seizures or toxicity. Thus, the monitoring and precise measuring of PHT concentrations in patients is crucial. This study develops and validates an LC-MS/MS method for the measurement of phenytoin concentrations in different body compartments (i.e., human brain dialysate, blood, and saliva and compares it with a formerly developed GC-MS method that measures PHT in the same biological matrices. The two methods are evaluated and compared based on their analytical performance, appropriateness to analyze human biological samples, including corresponding extraction and cleanup procedures, and their validation according to ISO 17025/FDA Guidance for Industry. The LC-MS/MS method showed a higher performance compared with the GC-MS method. The LC-MS/MS was more sensitive, needed a smaller sample volume (25 µL and less chemicals, was less time consuming (cleaning up, sample preparation, and analysis, and resulted in a better LOD ( 0.995 for all tested matrices (blood, saliva, and dialysate. For larger sample numbers as in pharmacokinetic/pharmacodynamic studies and for bedside as well as routine analyses, the LC-MS/MS method offers significant advantages over the GC-MS method.

  15. An On-Target Desalting and Concentration Sample Preparation Protocol for MALDI-MS and MS/MS Analysis

    DEFF Research Database (Denmark)

    Zhang, Xumin; Wang, Quanhui; Lou, Xiaomin

    2012-01-01

    2DE coupled with MALDI-MS is one of the most widely used and powerful analytic technologies in proteomics study. The MALDI sample preparation method has been developed and optimized towards the combination of simplicity, sample-cleaning, and sample concentration since its introduction. Here we...... present a protocol of the so-called Sample loading, Matrix loading, and on-target Wash (SMW) method which fulfills the three criteria by taking advantage of the AnchorChip™ targets. Our method is extremely simple and no pre-desalting or concentration is needed when dealing with samples prepared from 2DE...

  16. Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS.

    Science.gov (United States)

    Chen, Guan-Yuan; Chao, Hsi-Chun; Liao, Hsiao-Wei; Tsai, I-Lin; Kuo, Ching-Hua

    2017-12-15

    Glutamine, glutamate and glutathione are key modulators of excessive oxidative stress in tumor cells. In this study, we developed a rapid and accurate HILIC-MS/MS method to simultaneously determine concentrations of cellular glutamine, glutamate and glutathione. A bared silica HILIC column was employed to analyze these polar metabolites. The LC-MS parameters were optimized to achieve high sensitivity and selectivity. The analysis can be completed within 4 min under optimal conditions. The method was validated in terms of accuracy, precision, and linearity. Intra-day (n = 9) precision was within 2.68-6.24% among QCs. Inter-day precision (n = 3) was below 12.4%. The method accuracy was evaluated by the recovery test, and the accuracy for three analytes were between 91.6 and 110%. The developed method was applied to study antioxidant function of GLS2 in non-small cell lung cancer cells. Changes in concentrations of glutamine, glutamate and glutathione revealed that the overexpression of GLS2 could effectively decrease oxidative stress. In summary, this study developed a rapid HILIC-MS/MS method for quantification of GLS2-related metabolites that could facilitate elucidation of the role of GLS2 in tumor development. Copyright © 2017. Published by Elsevier Inc.

  17. Determination of 48 fragrance allergens in toys using GC with ion trap MS/MS.

    Science.gov (United States)

    Lv, Qing; Zhang, Qing; Li, Wentao; Li, Haiyu; Li, Pi; Ma, Qiang; Meng, Xianshuang; Qi, Meiling; Bai, Hua

    2013-11-01

    This paper presents a method for the simultaneous determination of 48 fragrance allergens in four types of toys (plastic toys, play clays, plush toys, and paper toys) based on GC with ion trap MS/MS. Compared with single-stage MS, MS/MS is superior in terms of the qualification and quantification of a large range of compounds in complicated matrices. Procedures for extraction and purification were optimized for each toy type. The method proved to be linear over a wide range of concentrations for all analytes with correlation coefficients between 0.9768 and 0.9999. Validation parameters, namely, LODs and LOQs, ranged from 0.005-5.0 and from 0.02-20 mg/kg, respectively. Average recoveries of target compounds (spiked at three concentration levels) were in the range of 79.5-109.1%. Intraday and interday repeatabilities of the proposed method varied from 0.7-10.5% and from 3.1-13.4%, respectively. The proposed method was used to monitor fragrance allergens in commercial toy products. Our findings indicate that this method is an accurate and effective technique for analyzing fragrance allergens in materials composed of complex components. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Lung Cancer Serum Biomarker Discovery Using Label Free LC-MS/MS

    Science.gov (United States)

    Zeng, Xuemei; Hood, Brian L.; Zhao, Ting; Conrads, Thomas P.; Sun, Mai; Gopalakrishnan, Vanathi; Grover, Himanshu; Day, Roger S.; Weissfeld, Joel L.; Wilson, David O.; Siegfried, Jill M.; Bigbee, William L.

    2011-01-01

    Introduction Lung cancer remains the leading cause of cancer-related death with poor survival due to the late stage at which lung cancer is typically diagnosed. Given the clinical burden from lung cancer, and the relatively favorable survival associated with early stage lung cancer, biomarkers for early detection of lung cancer are of important potential clinical benefit. Methods We performed a global lung cancer serum biomarker discovery study using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in a set of pooled non-small cell lung cancer (NSCLC) case sera and matched controls. Immunoaffinity subtraction was used to deplete the top most abundant serum proteins; the remaining serum proteins were subjected to trypsin digestion and analyzed in triplicate by LC-MS/MS. The tandem mass spectrum data were searched against the human proteome database and the resultant spectral counting data were used to estimate the relative abundance of proteins across the case/control serum pools. The spectral counting derived abundances of some candidate biomarker proteins were confirmed with multiple reaction monitoring MS assays. Results A list of 49 differentially abundant candidate proteins was compiled by applying a negative binomial regression model to the spectral counting data (pbiomarkers with statistically significant differential abundance across the lung cancer case/control pools which, when validated, could improve lung cancer early detection. PMID:21304412

  19. Development and validation of analytical method for Naftopidil in human plasma by LC–MS/MS

    Directory of Open Access Journals (Sweden)

    Pritam S. Jain

    2015-09-01

    Full Text Available A highly sensitive and simple high-performance liquid chromatographic–tandem mass spectrometric (LC–MS-MS assay is developed and validated for the quantification of Naftopidil in human plasma. Naftopidil is extracted from human plasma by methyl tertiary butyl ether and analyzed using a reversed-phase gradient elution on a discovery C 18 5 μ (50 × 4.6 column. A methanol: 2 mM ammonium formate (90:10 as mobile phase, is used and detection was performed by MS using electrospray ionization in positive mode. Propranolol is used as the internal standard. The lower limits of quantification are 0.495 ng/mL. The calibration curves are linear over the concentration range of 0.495–200.577 ng/mL of plasma for each analyte. This novel LC–MS-MS method shows satisfactory accuracy and precision and is sufficiently sensitive for the performance of pharmacokinetic studies in humans.

  20. SWATH Mass Spectrometry Performance Using Extended Peptide MS/MS Assay Libraries*

    Science.gov (United States)

    Wu, Jemma X.; Song, Xiaomin; Pascovici, Dana; Zaw, Thiri; Care, Natasha; Krisp, Christoph; Molloy, Mark P.

    2016-01-01

    The use of data-independent acquisition methods such as SWATH for mass spectrometry based proteomics is usually performed with peptide MS/MS assay libraries which enable identification and quantitation of peptide peak areas. Reference assay libraries can be generated locally through information dependent acquisition, or obtained from community data repositories for commonly studied organisms. However, there have been no studies performed to systematically evaluate how locally generated or repository-based assay libraries affect SWATH performance for proteomic studies. To undertake this analysis, we developed a software workflow, SwathXtend, which generates extended peptide assay libraries by integration with a local seed library and delivers statistical analysis of SWATH-quantitative comparisons. We designed test samples using peptides from a yeast extract spiked into peptides from human K562 cell lysates at three different ratios to simulate protein abundance change comparisons. SWATH-MS performance was assessed using local and external assay libraries of varying complexities and proteome compositions. These experiments demonstrated that local seed libraries integrated with external assay libraries achieve better performance than local assay libraries alone, in terms of the number of identified peptides and proteins and the specificity to detect differentially abundant proteins. Our findings show that the performance of extended assay libraries is influenced by the MS/MS feature similarity of the seed and external libraries, while statistical analysis using multiple testing corrections increases the statistical rigor needed when searching against large extended assay libraries. PMID:27161445

  1. Comparison of Proteins in Whole Blood and Dried Blood Spot Samples by LC/MS/MS

    Science.gov (United States)

    Chambers, Andrew G.; Percy, Andrew J.; Hardie, Darryl B.; Borchers, Christoph H.

    2013-09-01

    Dried blood spot (DBS) sampling methods are desirable for population-wide biomarker screening programs because of their ease of collection, transportation, and storage. Immunoassays are traditionally used to quantify endogenous proteins in these samples but require a separate assay for each protein. Recently, targeted mass spectrometry (MS) has been proposed for generating highly-multiplexed assays for biomarker proteins in DBS samples. In this work, we report the first comparison of proteins in whole blood and DBS samples using an untargeted MS approach. The average number of proteins identified in undepleted whole blood and DBS samples by liquid chromatography (LC)/MS/MS was 223 and 253, respectively. Protein identification repeatability was between 77 %-92 % within replicates and the majority of these repeated proteins (70 %) were observed in both sample formats. Proteins exclusively identified in the liquid or dried fluid spot format were unbiased based on their molecular weight, isoelectric point, aliphatic index, and grand average hydrophobicity. In addition, we extended this comparison to include proteins in matching plasma and serum samples with their dried fluid spot equivalents, dried plasma spot (DPS), and dried serum spot (DSS). This work begins to define the accessibility of endogenous proteins in dried fluid spot samples for analysis by MS and is useful in evaluating the scope of this new approach.

  2. Gas Chromatography (GC) – Mass Spectrometry (MS) analysis of ...

    African Journals Online (AJOL)

    MS. Glycosides, phenolics and alkaloids were indicated in large amount; eugenols and terpenes were also present while saponin, tannins and steroid were absent. Major components detected by GC-MS were 2-Hydroxy hexadecanoic acid ...

  3. Method-MS, final report 2010

    DEFF Research Database (Denmark)

    Skipperud, Lindis; Popic, Jelena M.; Roos, Per

    applied to the determination of the lighter stable element isotopes and radioactive isotopes in geological studies, the techniques can equally well be applied to the measurement of activity concentrations of long-lived low-level radionuclides in various samples using “isotope dilution” methods...... such as those applied in inductively coupled plasma mass spectrometry (ICP-MS). Due to the low specific activity of long-lived radionuclides, many of these are more conveniently detected using mass spectrometric techniques. Mass spectrometry also enables the individual determination of Pu-239 and Pu-240, which...

  4. Analysis of benzodiazepines and their metabolites using DBS cards and LC-MS/MS.

    Science.gov (United States)

    Lee, Heesang; Park, Yujin; Jo, Jiyeong; In, Sangwhan; Park, Yonghoon; Kim, Eunmi; Pyo, Jaesung; Choe, Sanggil

    2015-10-01

    Dried Blood Spot (DBS) has been used a blood extraction method for inherited metabolic disorder screening since 1960s. With introduction of LC-MS/MS, not only DBS could be used to analysis drugs in small blood volume, but in various fields, such as toxicology, drug therapeutic monitoring, drug diagnostic screening, and illicit drugs. In toxicology field, many drugs (e.g. benzodiazepines, acetaminophen, small molecule drugs) have been tested with DBS. Compared with earlier blood extraction methods (SPE and LLE), DBS has lots of advantages; lower blood volume (less than 50μL), shorter analysis time caused by a more concise analysis procedure and lower cost. We optimized the DBS procedure and LC-MS/MS conditions for 18 benzodiazepines, seven benzodiazepine metabolites, and one z-drug (zolpidem) analysis in blood. 30μL of whole blood was spotted on FTA DMPK card C and dried for 2h in a desiccator. A 6-mm disk was punched and vortexed for 1min in a centrifuge tube with 300μL methanol/acetonitrile mixture (1:1, v/v). After evaporation, redissolved in 100μL mobile phase of LC-MS/MS and 5μL was injected. In the analysis for 26 target compounds in blood, all of the method validation parameters - LLOD, LLOQ, accuracy (intra- and inter-assay), and precision (intra- and inter-assay) - were satisfied with method validation criteria, within 15%. The results of matrix effect, recovery, and process efficiency were good. We developed a fast and reliable sample preparation method using DBS for 26 benzodiazepines, benzodiazepine metabolites, and z-drug (zolpidem). Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  5. A UHPLC-MS/MS method for profiling multifunctional steroids in human hair.

    Science.gov (United States)

    Dong, Zhen; Wang, Caihong; Zhang, Jinlan; Wang, Zhe

    2017-08-01

    It is important to profile steroids in many physiological and pathological processes. Recently, hair has been used for the long-term measurement of endogenous steroid hormones. Analyzing hair has advantages of being noninvasive and time sequential compared with other bio-specimens. Liquid chromatography-mass spectrometry (LC-MS) techniques have been widely used over the past decades; however, it is challenging to profile estrogens in hair by LC-MS, and more comprehensive steroid profiling is required. In this paper, an ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to simultaneously profile 28 multifunctional steroids, including corticosteroids (n = 6), estrogens (n = 13), androgens (n = 5) and progestogens (n = 4), in human scalp hair in a single run. To optimize the sample preparation procedure, we evaluated extraction time, post-incubation purification and hair fragment length; 30 mg hair samples were washed with hexane, cut into 5 mm pieces and incubated in methanol for 18 h at 25 °C. Methanol extraction derivatized using Girard P and dansyl chloride reagent was analyzed within 25 min using an automated injection program combined with a diverter valve switch and step analysis (AIDSA). The method was well validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect and recovery, and was successfully applied to a steroid profile from male and female hairs. Significant differences were observed between genders. In addition, steroids showed a declining trend from the proximal to more distal hair segments; thus, care should be taken when obtaining hair samples for analysis to account for this difference in steroid levels along the length of hair. Graphical Abstract The workflow of the estabished UHPLC-MS/MS method.

  6. Comparison of LC-ESI-MS and GC-MS for the Analysis of a Synthetic Tabun Sample

    National Research Council Canada - National Science Library

    D'Agostino, Paul

    2003-01-01

    Packed capillary LC-ESI-MS and capillary column GC-MS were compared for the analysis of a synthetic tabun sample as each method has advantages for the analysis of samples containing chemical warfare...

  7. Application of LC–MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids

    Directory of Open Access Journals (Sweden)

    Jamshed Haneef

    2013-10-01

    Full Text Available Liquid chromatography tandem mass chromatography (LC–MS/MS is an important hyphenated technique for quantitative analysis of drugs in biological fluids. Because of high sensitivity and selectivity, LC–MS/MS has been used for pharmacokinetic studies, metabolites identification in the plasma and urine. This manuscript gives comprehensive analytical review, focusing on chromatographic separation approaches (column packing materials, column length and mobile phase as well as different acquisition modes (SIM, MRM for quantitative analysis of glucocorticoids and stimulants. This review is not meant to be exhaustive but rather to provide a general overview for detection and confirmation of target drugs using LC–MS/MS and thus useful in the doping analysis, toxicological studies as well as in pharmaceutical analysis. Keywords: LC–MS/MS, Ionization techniques, Glucocorticoids, Stimulants, Hyphenated techniques, Biological fluid

  8. Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum

    DEFF Research Database (Denmark)

    Søeborg, Tue; Frederiksen, Hanne; Johannsen, Trine Holm

    2017-01-01

    An isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of the ten steroid metabolites dehydroepiandrosterone sulfate (DHEAS), progesterone, 17α-hydroxyprogesterone (17-OHP), Δ4-androstenedione (Adione), corticosterone, 11-deoxycortisol, cortisol, cortisone, testosterone (T...

  9. Short Communication: Testosterone Measured with an Automatic Immunoassay Compares Reasonbly Well to Results Obtained by LC-MS/MS

    DEFF Research Database (Denmark)

    Knudsen, Cindy Søndersø; Højskov, Carsten Schriver; Møller, Holger Jon

    2016-01-01

    Background: Previous studies have reported problems measuring testosterone with immunological assays. Here we explore an automatic second generation immunoassay compared to a LC-MS/MS method. Methods: We collected blood samples from 76 women and measured testosterone, progesterone, gender...... hormonebinding globulin (SHBG), and albumin employing Cobas e601/c501. Testosterone, androstenedione (andro), dehydroepiandrosterone sulphate (DHEAS), and 17-hydroxyprogesterone (17-OHP) concentrations were measured employing LC-MS/MS. We evaluated the difference between testosterone measured by the two methods...... for the difference between results obtained by the two methods and the sample concentration of DHEAS and andro: Diff (Cobas e601 - LC-MS/MS) = 0.116 x DHEAS - 0.396, r = 0.84 and Diff (Cobas e601 - LC-MS/MS) = 0.08 andro - 0.380, r = 0.58. No statistically significant interference was observed for progesterone, 17...

  10. STS-47 MS Davis and MS Jemison conduct LBNP experiment in the SLJ module

    Science.gov (United States)

    1992-01-01

    At the aft end of the Spacelab Japan (SLJ) science module, STS-47 Mission Specialist (MS) N. Jan Davis (foreground) readies Rack 9 Automatic Blood Pressure System (ABPS) controls as MS Mae C. Jemison, inside the cylindrical fabric lower body negative pressure (LBNP) device, waits for the LBNP experiment to begin. LBNP device is sealed around Jemison's waist. It is attached to the SLJ floor and has a controller that operates a pump to change the pressure inside. Davis will monitor Jemison's pulse rate, blood pressure, and cardiac dimensions and functions.

  11. Comparison of CE-MS and LC-MS Analyses of Avian Eggshell Matrix Proteins

    Czech Academy of Sciences Publication Activity Database

    Mikšík, Ivan; Sedláková, Pavla; Mikulíková, Kateřina; Eckhardt, Adam; Kašička, Václav

    2008-01-01

    Roč. 67, Suppl.1 (2008), s. 89-96 ISSN 0009-5893 R&D Projects: GA MŠk(CZ) 1M0510; GA ČR(CZ) GA203/06/1044; GA ČR(CZ) GA203/05/2539 Institutional research plan: CEZ:AV0Z50110509; CEZ:AV0Z40550506 Keywords : eggshell proteins * CE-MS * HPLC-MS Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 1.312, year: 2008

  12. Quantitative analysis of methyl and propyl parabens in neonatal DBS using LC-MS/MS.

    Science.gov (United States)

    Yakkundi, Shirish; Mulla, Hussain; Pandya, Hitesh; Turner, Mark A; McElnay, James

    2016-06-01

    Excipients are used to overcome the chemical, physical and microbiological challenges posed by developing formulated medicines. Both methyl and propyl paraben are commonly used in pediatric liquid formulations. There is no data on systemic exposure to parabens in neonates. The European Study of Neonatal Exposure to Excipients project has investigated this. Results & methodology: DBS sampling was used to collect opportunistic blood samples. Parabens were extracted from the DBS and analyzed using a validated LC-MS/MS assay. The above assay was applied to analyze neonatal DBS samples. The blood concentrations of parabens in neonates confirm systemic exposure to parabens following administration of routine medicines.

  13. Analysis of pesticide residues in strawberries and soils by GC-MS/MS, LC-MS/MS and two-dimensional GC-time-of-flight MS comparing organic and integrated pest management farming.

    Science.gov (United States)

    Fernandes, Virgínia C; Lehotay, Steven J; Geis-Asteggiante, Lucía; Kwon, Hyeyoung; Mol, Hans G J; van der Kamp, Henk; Mateus, Nuno; Domingues, Valentina F; Delerue-Matos, Cristina

    2014-01-01

    This study analysed 22 strawberry and soil samples after their collection over the course of 2 years to compare the residue profiles from organic farming with integrated pest management practices in Portugal. For sample preparation, we used the citrate-buffered version of the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. We applied three different methods for analysis: (1) 27 pesticides were targeted using LC-MS/MS; (2) 143 were targeted using low pressure GC-tandem mass spectrometry (LP-GC-MS/MS); and (3) more than 600 pesticides were screened in a targeted and untargeted approach using comprehensive, two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOF-MS). Comparison was made of the analyses using the different methods for the shared samples. The results were similar, thereby providing satisfactory confirmation of both similarly positive and negative findings. No pesticides were found in the organic-farmed samples. In samples from integrated pest management practices, nine pesticides were determined and confirmed to be present, ranging from 2 µg kg(-1) for fluazifop-p-butyl to 50 µg kg(-1) for fenpropathrin. Concentrations of residues in strawberries were less than European maximum residue limits.

  14. Moessbauer spectrometer MsAa-3

    International Nuclear Information System (INIS)

    Gornicki, R.; Blachowski, A.; Ruebenbauer, K.

    2007-01-01

    The paper is aimed at the description of the newly developed Moessbauer spectrometer MsAa-3. The spectrometer MsAa-3 consists of a high quality γ--ray spectrometer including either a proportional gas detector head or a scintillation detector head, a transducer driving system including the transducer, data storage system, and data communication system based on the TCP/IP protocol. Additionally, the Michelson-Morley interferometer is provided for precise calibration of the transducer velocity. The spectrometer is equipped with an integrated simple temperature controller. All the essential functions are remotely controlled over the TCP/IP link allowing for the spectrometer set-up as the stand-alone unit in the computer network, e.g. on the Internet. External γ-ray detectors or external complete nuclear blocks could be used as well. The spectrometer is equipped with software allowing for setting all the functions, to perform on-line control, and retrieve data. The Moessbauer data processing software MOSGRAF is enclosed as well. The latter software allows for the calculation of the variety of velocity reference functions. (authors)

  15. ms_lims, a simple yet powerful open source laboratory information management system for MS-driven proteomics.

    Science.gov (United States)

    Helsens, Kenny; Colaert, Niklaas; Barsnes, Harald; Muth, Thilo; Flikka, Kristian; Staes, An; Timmerman, Evy; Wortelkamp, Steffi; Sickmann, Albert; Vandekerckhove, Joël; Gevaert, Kris; Martens, Lennart

    2010-03-01

    MS-based proteomics produces large amounts of mass spectra that require processing, identification and possibly quantification before interpretation can be undertaken. High-throughput studies require automation of these various steps, and management of the data in association with the results obtained. We here present ms_lims (http://genesis.UGent.be/ms_lims), a freely available, open-source system based on a central database to automate data management and processing in MS-driven proteomics analyses.

  16. VOC identification and inter-comparison from laboratory biomass burning using PTR-MS and PIT-MS

    Science.gov (United States)

    C. Warneke; J. M. Roberts; P. Veres; J. Gilman; W. C. Kuster; I. Burling; R. Yokelson; J. A. de Gouw

    2011-01-01

    Volatile organic compounds (VOCs) emitted from fires of biomass commonly found in the southeast and southwest U.S. were investigated with PTR-MS and PIT-MS, which are capable of fast measurements of a large number of VOCs. Both instruments were calibrated with gas standards and mass dependent calibration curves are determined. The sensitivity of the PIT-MS linearly...

  17. Analytical Method Details (MS): SE57_MS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available TQD, UPLC (Waters) UPLC-QTOF-MS ESI N.A. All the solutions of authentic....1% formic acid), with a 2.5 min cycle in the isocratic mode. The ionized authentic compounds were detected ... optimal CV and CE were determined automatically for 497 of these compounds using the software QuanOptimize

  18. Potential Health Benefits and Metabolomics of Camel Milk by GC-MS and ICP-MS.

    Science.gov (United States)

    Ahamad, Syed Rizwan; Raish, Mohammad; Ahmad, Ajaz; Shakeel, Faiyaz

    2017-02-01

    None of the research reports reveals the metabolomics and elemental studies on camel milk. Recent studies showed that camel milk possesses anticancer and anti-inflammatory activity. Metabolomics and elemental studies were carried out in camel milk which showed us the pathways and composition that are responsible for the key biological role of camel milk. Camel milk was dissolved in methanol and chloroform fraction and then vortexed and centrifuged. Both the fractions were derivatized by N,O-bis-(trimethylsilyl)trifluoroacetamide (BSTFA) and TMCS after nitrogen purging and analyzed by GC-MS. Camel milk was also analyzed by ICP-MS after microwave digestion. We found that higher alkanes and fatty acids are present in the chloroform fraction and amino acids, sugars and fatty acid derivatives are present in aqueous fractions. All the heavy metals like As, Pb, Cd, Co, Cu, and Ni were in the safe limits in terms of maximum daily intake of these elements. Na, K, Mg, and Ca were also present in the safe limits in terms of maximum daily intake of these elements. These results suggested that the camel milk drinking is safe and there is no health hazard. The present data of GC-MS and ICP-MS correlate the activities related to camel milk.

  19. [LC-MS/MS analysis of determination of strychnine and brucine in formaldehyde fixed tissue].

    Science.gov (United States)

    Zhan, Lan-fen; Liu, Ming-dong; Yan, You-yi; Ye, Yi; Wang, Wei; Wang, Zhi-hui; Zhao, Jun-hong; Liao, Lin-chuan

    2012-10-01

    To establish a method for determination of strychnine and brucine in formaldehyde fixed tissue by LC-MS/MS analysis. The samples were pretreated with solid phase extraction using SCX cartridges and separated on SB-C18 column with mobile phase 0.1% formic acid : 0.1% formic acid-acetonitrile (75:25). Electrospray ionization (ESI) source was utilized and operated in positive ion mode. Multiple reactions monitoring (MRM) mode was applied. External standard method was applied for quantitation. The chromatographic separation of strychnine and brucine in formaldehyde fixed nephritic and hepatic tissues resulted successfully. The standard curve was linear in the range of 0.002-2.0 microg/g for strychnine and brucine in formaldehyde fixed tissues, and the correlation coefficient was more than 0.996. The limits of detection (LOD) of strychnine and brucine in nephritic tissues were 0.06ng/g and 0.03 ng/g, respectively. The LOD of both chemicals were 0.3 ng/g in hepatic tissues. The extraction recovery rate was more than 74.5%. The precision of intra-day and inter-day were both less than 8.2%. Strychnine and brucine can be sensitive to be determined in formaldehyde fixed tissue by LC-MS/MS analysis. It can be applied in the forensic toxicological analysis.

  20. UP-HILIC-MS/MS to Determine the Action Pattern of Penicillium sp. Dextranase

    Science.gov (United States)

    Yi, Lin; Sun, Xue; Du, Kenze; Ouyang, Yilan; Wu, Chengling; Xu, Naiyu; Linhardt, Robert J.; Zhang, Zhenqing

    2015-07-01

    Investigation of the action pattern of enzymes acting on carbohydrates is challenging, as both the substrate and the digestion products are complex mixtures. Dextran and its enzyme-derived oligosaccharides are widely used for many industrial applications. In this work, a new method relying on ultra-performance hydrophilic interaction liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UP-HILIC- Q/TOF-MS/MS) was developed to analyze a complex mixture of dextran oligosaccharide products to determine the action pattern of dextranase. No derivatization of oligosaccharides was required and the impact of the α- and β-configurations of the native oligosaccharides on the chromatographic separation was eliminated. The 1→6, 1→3, 1→4 backbone linkages and the branch linkages of these oligosaccharides were all distinguished from diagnostic ions in their MS/MS spectra, including fragments corresponding to 0,2A, 0,3A, 0,4A, B-H2O, 2,5A, and 3,5A. The sequences of the oligosaccharide products were similarly established. Thus, the complex oligosaccharide mixtures in dextran digestion products were profiled and identified using this method. The more enzyme-resistant structures in dextran were established using much less sample, labor, time, and uncertainty than in previous studies. This method provides an efficient, sensitive, and straightforward way to monitor the entire process of digestion, establish the action pattern of the dextranase from Penicillium sp., and to support the proper industrial application of dextranase.

  1. Highly sensitive quantification of key regulatory oxysterols in biological samples by LC-ESI-MS/MS.

    Science.gov (United States)

    Honda, Akira; Yamashita, Kouwa; Hara, Takashi; Ikegami, Tadashi; Miyazaki, Teruo; Shirai, Mutsumi; Xu, Guorong; Numazawa, Mitsuteru; Matsuzaki, Yasushi

    2009-02-01

    We describe a highly sensitive and specific method for the quantification of key regulatory oxysterols in biological samples. This method is based upon a stable isotope dilution technique by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After alkaline hydrolysis of human serum (5 microl) or rat liver microsomes (1 mg protein), oxysterols were extracted, derivatized into picolinyl esters, and analyzed by LC-MS/MS using the electrospray ionization mode. The detection limits of the picolinyl esters of 4beta-hydroxycholesterol, 7alpha-hydroxycholesterol, 22R-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 27-hydroxycholesterol, and 24S,25-epoxycholesterol were 2-10 fg (5-25 amol) on-column (signal-to-noise ratio = 3). Reproducibilities and recoveries of these oxysterols were validated according to one-way layout and polynomial equation, respectively. The variances between sample preparations and between measurements by this method were calculated to be 1.8% to 12.7% and 2.9% to 11.9%, respectively. The recovery experiments were performed using rat liver microsomes spiked with 0.05 ng to 12 ng of oxysterols, and recoveries of the oxysterols ranged from 86.7% to 107.3%, with a mean recovery of 100.6%. This method provides reproducible and reliable results for the quantification of oxysterols in small amounts of biological samples.

  2. HPLC/QTOF-MS/MS application to investigate phenolic constituents from Ficus pandurata H. aerial roots.

    Science.gov (United States)

    Zhang, Xiaoping; Lv, Huiqing; Li, Zuguang; Jiang, Kezhi; Lee, Maw-Rong

    2015-06-01

    Ficus pandurata H. aerial roots are used as a traditional Chinese medicine for the treatment of uarthritis, indigestion and hyperuricemia. However, the bioactive constituents responsible for the pharmacological effects of F. pandurata H. are unclear. A simple and efficient HPLC/QTOF-MS/MS (high-performance liquid chromatography/electrospray ionization with quadrupole time-of-flight tandem mass spectrometry) method was established to detect and identify active constituents in the n-butanol extract of F. pandurata H. aerial roots. Chemical constituents were separated and investigated by HPLC/QTOF-MS/MS in the negative-ion mode. Thirty-seven compounds, including hydroxycinnamic acid derivatives, hydroxybenzoic acid derivatives, hydroquinone glycosides, flavonoid glycosides, etc., were identified or tentatively characterized in the n-butanol extract of F. pandurata H. aerial roots by comparing the UV spectra, accurate mass spectra and fragmentation pathways and retrieving the reference literatures. Moreover, the flavonoid trisaccharides and hydroxybenzoic acid derivatives were tentatively characterized in F. pandurata H. for the first time. The analytical tool used here is very valuable in the rapid separation and identification of the multiple and minor constituents in the n-butanol extract of F. pandurata H. aerial roots. Copyright © 2014 John Wiley & Sons, Ltd.

  3. Simultaneous determination of 38 veterinary antibiotic residues in raw milk by UPLC-MS/MS.

    Science.gov (United States)

    Han, R W; Zheng, N; Yu, Z N; Wang, J; Xu, X M; Qu, X Y; Li, S L; Zhang, Y D; Wang, J Q

    2015-08-15

    A selective and rapid method has been developed to determine, simultaneously, 38 veterinary antibiotic residues in raw milk by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). One milliliter of raw milk was diluted with 0.5 mL water and 3 mL acetonitrile, then purified using an Oasis HLB cartridge. The eluates were evaporated by nitrogen drying and then reconstituted to 4 mL with water/acetonitrile (8:1) before being injected into the UPLC-MS/MS system. The results indicated recoveries of 68-118% for 14 β-lactams, 79-118% for eight quinolones, 71-106% for eight sulfonamides, 76-116% for four tetracyclines, 78-106% for three macrolides, and 88-103% for one lincosamides, with coefficients of variation less than 15% for intraday and interday precisions. The limit of quantification for all antibiotics was 0.03-10 μg kg(-1). This methodology was then applied to field-collected real raw milk samples and trace levels of four antibiotics were detected. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Mycotoxin production of Alternaria strains isolated from Korean barley grains determined by LC-MS/MS.

    Science.gov (United States)

    Nguyen, Thuong T T; Kim, Jueun; Jeon, Sun Jeong; Lee, Chul Won; Magan, Naresh; Lee, Hyang Burm

    2018-03-02

    Twenty-four Alternaria strains were isolated from barley grain samples. These strains were screened for the production of mycotoxins on rice medium using thin layer chromatography. All 24 strains produced at least one of the five mycotoxins (ALT, AOH, ATX-I, AME, and TeA). Three representative strains, namely EML-BLDF1-4, EML-BLDF1-14, and EML-BLDF1-18, were further analyzed using a new LC-MS/MS-based mycotoxin quantification method. This method was used to detect and quantify Alternaria mycotoxins. We used positive ion electrospray mass spectrometry with multiple reaction mode (MRM) for the simultaneous quantification of various Alternaria mycotoxins produced by these strains. Five Alternaria toxins (ALT, ATX-I, AOH, AME, and TeA) were detected and quantified. Sample preparation included methanol extraction, concentration, and injection into LC-MS/MS. Limit of detection ranged from 0.13 to 4μg/mL and limit of quantification ranged from 0.25 to 8μg/mL. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Integrated GC-MS and LC-MS plasma metabonomics analysis of ankylosing spondylitis.

    Science.gov (United States)

    Gao, Peng; Lu, Chen; Zhang, Fengxia; Sang, Ping; Yang, Dawei; Li, Xiang; Kong, Hongwei; Yin, Peiyuan; Tian, Jing; Lu, Xin; Lu, Aiping; Xu, Guowang

    2008-09-01

    Ankylosing spondylitis (AS) is a chronic inflammatory arthritis that predominantly affects the axial skeleton in adolescent patients. The natural history of the disease remains poorly characterized. In this study, we combined GC-MS and LC-MS techniques to evaluate the major metabolic changes in the plasma of AS patients in view of metabonomics. Univariate and multivariate analysis were employed for altered metabolite comparison and pattern recognition. Application of supervised partial least-squares discrminant analysis to either GC-MS or LC-MS data allowed accurate discrimination of AS patients from normal controls, demonstrating its potential diagnostic utilization. In addition, AS patients presented elevated plasma concentrations of proline, glucose, phosphate, urea, glycerol, phenylalanine and homocysteine but reduced levels of phosphocholines, tryptophan and a bipeptide - phenylalanyl-phenylalanine. In the context of their involved metabolic pathways, the identified metabolites were discussed accordingly. This investigation primarily proved that integrated chromatography-mass spectrometry and integrated uni- and multi-variate statistical analysis facilitated metabonomics to be a more promising tool in disease research.

  6. [Simultaneous determination of gestodene, etonogestrel and ethinylestradiol in plasma by LC-MS/MS following derivatization].

    Science.gov (United States)

    Liu, Xiao-Fen; Ding, Cun-Gang; Ge, Qing-Hua; Zhou, Zhen; Zhi, Xiao-Jin

    2010-01-01

    To establish a sensitive and specific method for simultaneous determination of gestodene, etonogestrel and ethinylestradiol in plasma by LC-MS/MS, plasma samples were extracted and derivatized before injection. An ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring (MRM). Norgestrel was chosen as internal standard and performed on a C18 (100 mm x 2.1 mm, 5 microm) column. The concentrations of gestodene, etonogestrel and ethinylestradiol were measured, using step-gradient mobile phase and step-gradient flow rate. The method was validated over the concentration range of 0.1-20 ng x mL(-1) for gestodene and etonogestrel and 0.01-2 ng x mL(-1) for ethinylestradiol, and showed excellent linearity. The intra- and inter-assay accuracy and precision were below 10.0% and recovery was 93.6%-110.9% over the three concentration levels evaluated. The method was applied in pharmacokinetic study of the compound gestodene patch and the compound etonogestrel patch in rabbits. The LC-MS/MS method was selective, accurate and sensitive, especially the LOQ were 100 pg x mL(-1) for gestodene and etonogestrel and 10 pg x mL(-1) for ethinylestradiol. The method was successfully applied in pharmacokinetic study for contraceptives.

  7. Long term anoxia in rainbow trout investigated by 2-DE and MS/MS

    DEFF Research Database (Denmark)

    Wulff, Tune; Jessen, Flemming; Roepstorff, P.

    2008-01-01

    Twenty-four hours of N-2 induced anoxia induced global perturbations on protein expression in rainbow trout hypodermal fibroblasts cell line. Anoxia was obtained by depleting the medium of O-2 by flushing with N-2, and protein changes were studied by 2-DE coupled with MS providing quantitative...

  8. Efficient analysis and extraction of MS/MS result data from Mascot™ result files

    Directory of Open Access Journals (Sweden)

    Sickmann Albert

    2005-12-01

    Full Text Available Abstract Background Mascot™ is a commonly used protein identification program for MS as well as for tandem MS data. When analyzing huge shotgun proteomics datasets with Mascot™'s native tools, limits of computing resources are easily reached. Up to now no application has been available as open source that is capable of converting the full content of Mascot™ result files from the original MIME format into a database-compatible tabular format, allowing direct import into database management systems and efficient handling of huge datasets analyzed by Mascot™. Results A program called mres2x is presented, which reads Mascot™ result files, analyzes them and extracts either selected or all information in order to store it in a single file or multiple files in formats which are easier to handle downstream of Mascot™. It generates different output formats. The output of mres2x in tab format is especially designed for direct high-performance import into relational database management systems using native tools of these systems. Having the data available in database management systems allows complex queries and extensive analysis. In addition, the original peak lists can be extracted in DTA format suitable for protein identification using the Sequest™ program, and the Mascot™ files can be split, preserving the original data format. During conversion, several consistency checks are performed. mres2x is designed to provide high throughput processing combined with the possibility to be driven by other computer programs. The source code including supplement material and precompiled binaries is available via http://www.protein-ms.de and http://sourceforge.net/projects/protms/. Conclusion The database upload allows regrouping of the MS/MS results using a database management system and complex analyzing queries using SQL without the need to run new Mascot™ searches when changing grouping parameters.

  9. Segmental analysis of amphetamines in hair using a sensitive UHPLC-MS/MS method.

    Science.gov (United States)

    Jakobsson, Gerd; Kronstrand, Robert

    2014-06-01

    A sensitive and robust ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for quantification of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylenedioxy methamphetamine in hair samples. Segmented hair (10 mg) was incubated in 2M sodium hydroxide (80°C, 10 min) before liquid-liquid extraction with isooctane followed by centrifugation and evaporation of the organic phase to dryness. The residue was reconstituted in methanol:formate buffer pH 3 (20:80). The total run time was 4 min and after optimization of UHPLC-MS/MS-parameters validation included selectivity, matrix effects, recovery, process efficiency, calibration model and range, lower limit of quantification, precision and bias. The calibration curve ranged from 0.02 to 12.5 ng/mg, and the recovery was between 62 and 83%. During validation the bias was less than ±7% and the imprecision was less than 5% for all analytes. In routine analysis, fortified control samples demonstrated an imprecision <13% and control samples made from authentic hair demonstrated an imprecision <26%. The method was applied to samples from a controlled study of amphetamine intake as well as forensic hair samples previously analyzed with an ultra high performance liquid chromatography time of flight mass spectrometry (UHPLC-TOF-MS) screening method. The proposed method was suitable for quantification of these drugs in forensic cases including violent crimes, autopsy cases, drug testing and re-granting of driving licences. This study also demonstrated that if hair samples are divided into several short segments, the time point for intake of a small dose of amphetamine can be estimated, which might be useful when drug facilitated crimes are investigated. Copyright © 2014 John Wiley & Sons, Ltd.

  10. Pyrolysis characteristic of tobacco stem studied by Py-GC/MS, TG-FTIR, and TG-MS

    OpenAIRE

    Bei Liu; You-Ming Li; Shu-Bin Wu; Yan-Heng Li; Shan-Shan Deng; Zheng-Lin Xia

    2013-01-01

    Pyrolysis characteristics and mechanism of tobacco stem were studied by pyrolysis coupled with gas chromatography/mass spectrometry (Py-GC/MS), thermogravimetric analyzer coupled with Fourier transform infrared spectrometry, and mass spectrometry (TG-FTIR and TG-MS) techniques. The composition of evolved volatiles from fast pyrolysis of tobacco stem was determined by Py-GC/MS analysis, and the evolution patterns of the major products were investigated by TG-FTIR and TG-MS. Py-GC/MS data indic...

  11. Adaptation of GAMANAL to MS-DOS

    International Nuclear Information System (INIS)

    Nagy, S.

    1990-04-01

    The public domain gamma spectrum analysis programme GAMANAL was adapted for use with personal computers running with MS-DOS or equivalent operating system. The original command structure of GAMANAL, and its algorithms were kept unchanged; the facilities to read spectra from different multichannel analyzer formats were added. In the completed PC version, GAMANAL runs in a batch mode, without graphics presentation or menu-oriented interaction with the operator. The operation and performance of the PC version of GAMANAL was compared with some commercial software, in respect to peak sensitivity, multiplet resolution, and accuracy of area determination. The study indicates that the GAMANAL analysis yields results as good or better than other programmes. 4 refs

  12. Identification of ractopamine glucuronides and determination of bioactive ractopamine residues and its metabolites in food animal urine by ELISA, LC-MS/MS and GC-MS.

    Science.gov (United States)

    Jiang, Xiao-Fei; Zhu, Yue-Hui; Liu, Xiao-Yun

    2014-01-01

    Ractopamine glucuronides have been identified in cattle urine sampled by LC-MS/MS. An ELISA method, which was capable of specifically determining (1R, 3R)-ractopamine stereoisomer and its glucuronide metabolites, had more than 100% recovery with an acceptable coefficient of variation in the inter- and intra-assay variation tests for RR-ractopamine. The concentration levels of parent ractopamine and ractopamine glucuronide metabolites as the main components of total ractopamine in cattle and sheep urine showed similar depletion trends, in which the concentration curves increased and reached a climax during the feeding period, and then dropped quickly when entering the withdrawal period. Data from the three methods had very good pair-wise correlations. In the cattle urine samples, the correlation coefficient (R(2)) for parent ractopamine between the ELISA and the LC-MS/MS or GC-MS results were 0.93 or 0.92; R(2) values for parent ractopamine and total ractopamine data measured by LC-MS/MS and GC-MS were 0.9651 and 0.9677, respectively. All R(2) values for data gained from sheep urine samples were >0.95. The study indicated that the close levels of RR-ractopamine stereoisomer in cattle and sheep urine samples may imply the presence of a similar depletion pattern in other livestock, and thus would facilitate an accurate detection and management of ractopamine usage in food safety.

  13. Coupling of capillary zone electrophoresis to mass spectrometry (MS and MS/MS) via a nanoelectrospray interface for the characterisation of some β-agonists

    NARCIS (Netherlands)

    Guček, M.; Vreeken, R.J.; Verheij, E.R.

    1999-01-01

    Capillary zone electrophoresis (CZE) was coupled to mass spectrometry (MS) via a nanoelectrospray interface (nESI), using conductively coated tips (8 μm orifice) butted to the end of a fused-silica capillary. The CZE/nESI-MS system was carefully optimised to achieve a stable operation of the

  14. Aflatoxins contamination in Pakistani brown rice: a comparison of TLC, HPLC, LC-MS/MS and ELISA techniques.

    Science.gov (United States)

    Iqbal, Javed; Asghar, Muhammad Asif; Ahmed, Aftab; Khan, Mobeen Ahmed; Jamil, Khalid

    2014-12-01

    Advancement in the field of analytical food-chemistry has explored various experimental techniques for aflatoxins (AFs) quantification. The present study was aimed to compare four different techniques; thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC-MS/MS) and enzyme-linked immunosorbent assay (ELISA) for the analysis of aflatoxin B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) in brown rice (n = 120) being collected from Karachi, Pakistan. All the four assays provide precised, accurate and comparable results. However, some differences were observed. For instance, TLC, HPLC and LC-MS/MS methodologies offered the advantage of the quantification of individual toxins in contrast to ELISA technique. The contamination ranges of AFB₁/AFB₂ as determined by TLC, HPLC and LC-MS/MS were 1.18-9.97/0.59-1.52, 0.16-10.54/0.26-1.35 and 0.11-10.88/0.38-1.48 µg/kg, respectively. However, AFG₁ and AFG₂ were not detected in any tested samples. Furthermore, owing to low-detection limit and sensitivity, HPLC and LC-MS/MS methodologies have identified greater number of contaminated samples in comparison to TLC and ELISA techniques. The overall average results of total AFs as provided by HPLC (3.79 µg/kg) and LC-MS/MS (3.89 µg/kg) were found higher in comparison to TLC (3.68 µg/kg) and ELISA (3.70 µg/kg). On the basis of achieved results, it was concluded that TLC, HPLC, LC-MS/MS and ELISA techniques are valuable tool for the quantification of AFs in cereals and grains. Furthermore, HPLC and LC-MS/MS techniques offer an added advantage for the detection of AFs in diminutive levels.

  15. Cloud point extraction for analysis of antiretrovirals in human plasma by UFLC-ESI-MS/MS

    Directory of Open Access Journals (Sweden)

    Gabriel A. Hunzicker

    2015-12-01

    Full Text Available An analytical methodology based on cloud point extraction (CPE coupled to Ultra-Fast Liquid Chromatography and electrospray tandem mass spectrometry (UFLC-MS/MS was developed for analysis of Abacavir (ABC, Efavirenz (EFV, Lamivudine (3 TC and Nelfinavir (NFV in human plasma. It is the first time that CPE was used for extraction of antiretrovirals (ARV from plasma. The effects of relevant physic-chemical variables on analytical response of each ARV, including pH, surfactant concentration, equilibration time and temperature, were study and optimized; as well as its coupling to UFLC-ESI-MS/MS. Under optimized conditions, the resulting methodology was as follows: a 500 μL aliquot of human plasma was diluted with 2 mL deionized water in a 10 mL centrifuge tube. A 500 μL aliquot Triton X-114 5% w/v was added and homogenized using a vortex stirrer. The resulting cloudy solution was kept at 65 °C for 20 min for promoting the condensation of surfactant micelles. Then it was centrifuged at 3000 × g for 5 min for separation of the surfactant-rich phase. After discarding the aqueous supernatant, 400 μL ACN were added to the remaining surfactant rich phase and centrifuged in order to precipitate proteins and separate them. A 150 μL aliquot of the supernatant was transferred to 2 mL vial and further diluted with 400 μL deionized water. A 30 μL aliquot of the so-prepared solution was injected and analyzed into the UFLC-MS/MS. The method detection limits for ABC, EFV, 3 TC and NFV under optimized conditions were 31, 77, 57 and 21 ng mL−1, respectively. The RSD% for the studied analytes were <15%, except at the LOQ, which were <19%. Recovery values ranged from 81 to 107%. The proposed methodology was successfully applied for the analysis of ABC, EFV, 3 TC and NFV in human plasma within the concentration range of 43–6816, 125–4992, 81–3248 and 49–7904 ng mL−1, respectively. Under optimized working conditions the proposed

  16. Cannabinoids determination in oral fluid by SPME-GC/MS and UHPLC-MS/MS and its application on suspected drivers.

    Science.gov (United States)

    Anzillotti, Luca; Castrignanò, Erika; Strano Rossi, Sabina; Chiarotti, Marcello

    2014-12-01

    The confirmation of Δ9-tetrahydrocannabinol (THC) in oral fluid (OF) is an important issue for assessing Driving Under the Influence of Drugs (DUID). The aim of this research was to develop a highly sensitive method with minimal sample pre-treatment suitable for the analysis of small OF volumes (100 μL) for the confirmation of cannabinoids in DUID cases. Two methods were compared for the confirmation of THC in residual OF samples, obtained from a preliminary on-site screening with commercial devices. An ultra high performance LC-MS (UHPLC-MS/MS) method and an SPME-GC/MS method were hence developed. 100 μL of the residual mixture OF/preservative buffer or neat OF was simply added to 10 μL of THC-D3 (1 μg/mL) and submitted to the two different analyses: A - direct injection of 10 μL in UHPLC-MS/MS in positive electrospray ionisation (ESI) mode and B - sampling for 30 min with SPME (100 μm polydimethylsiloxane or PDMS fibre) and direct injection by desorption of the fibre in the GC injection port. The lowest limit of detection (LLOD) of THC was 2 ng/mL in UHPLC-MS/MS and 0.5 ng/mL in SPME-GC/MS. In addition, cannabidiol (CBD) and cannabinol (CBN) could be detected in GC/MS equipment at 2 ng/mL, whilst in UHPLC-MS/MS the LLOD was 20 ng/mL. Both methods were applied to 70 samples coming from roadside tests. By SPME-GC/MS analysis, THC was confirmed in 42 samples, whilst CBD was detected in 21 of them, along with CBN in 14 samples. THC concentrations ranged from traces below the lowest limit of quantification or LLOQ (2 ng/mL) up to 690 ng/mL. Copyright © 2014 Forensic Science Society. Published by Elsevier Ireland Ltd. All rights reserved.

  17. Tracing fungi secondary metabolites in Brazil nuts using LC-MS/MS.

    Science.gov (United States)

    Freitas-Silva, Otniel; de Lourdes, Maria; de Souza, Mendes; Venãncio, Armando

    2011-08-01

    This screening aimed to evaluate quantitatively the occurrence of fungal metabolites in Brazil nuts. Nuts were collected from Agroforest production areas in Amazon basin region. A total of 235 mycotoxins (including the most prominent ones) was screened by a multi-mycotoxin method based on HPLC-MS/MS. The recovery of metabolites by the method was between 56 and 136%. Fifteen mycotoxins were detected and quantified, in at least one sample; namely, aflatoxins (AFB(1), AFB(2), AFG(1), and AFM(1)), sterigmatocystin, methyl-sterigmatocystin, kojic acid, citrinin, cyclosporin A, cyclosporin C, cyclosporin D, cyclosporin H, rugulosin, alternariol-methylether and emodin. This is the first study dealing with the detection of the latter nine metabolites in Brazil nuts. Alternariol-methylether (from 0.75 to 3.2 µg/kg) was the only metabolite detected in all analyzed samples.

  18. Determination of phenolic compounds in Yucca gloriosa bark and root by LC-MS/MS.

    Science.gov (United States)

    Montoro, Paola; Skhirtladze, Alexandre; Bassarello, Carla; Perrone, Angela; Kemertelidze, Ether; Pizza, Cosimo; Piacente, Sonia

    2008-08-05

    On the basis of the biological activities shown by yuccaols and gloriosaols from Yucca schidigera and Yucca gloriosa, the content of yuccaols and gloriosaols in two different parts of Y. gloriosa (roots and bark), was determined for each single compound, and compared with phenolic determination in Y. schidigera bark, concluding that Y. gloriosa bark and roots are rich sources of phenolic derivatives structurally related to resveratrol. LC/ESIMS (liquid chromatography coupled to electrospray mass spectrometry) qualitative and an LC/ESIMS/MS (liquid chromatography coupled to tandem electrospray mass spectrometry) quantitative studies of the phenolic fraction of Y. gloriosa were performed. LC/ESIMS/MS multiple reaction monitoring (MRM) method previously described for yuccaols in Y. schidigera was applied and optimised for separation and determination of gloriosaols and yuccaols in Y. gloriosa. Due to the sensitivity and the repeatability of the assay, we suggest this method as suitable for industrial quality control of raw materials and final products.

  19. Current advances and strategies towards fully automated sample preparation for regulated LC-MS/MS bioanalysis.

    Science.gov (United States)

    Zheng, Naiyu; Jiang, Hao; Zeng, Jianing

    2014-09-01

    Robotic liquid handlers (RLHs) have been widely used in automated sample preparation for liquid chromatography-tandem mass spectrometry (LC-MS/MS) bioanalysis. Automated sample preparation for regulated bioanalysis offers significantly higher assay efficiency, better data quality and potential bioanalytical cost-savings. For RLHs that are used for regulated bioanalysis, there are additional requirements, including 21 CFR Part 11 compliance, software validation, system qualification, calibration verification and proper maintenance. This article reviews recent advances in automated sample preparation for regulated bioanalysis in the last 5 years. Specifically, it covers the following aspects: regulated bioanalysis requirements, recent advances in automation hardware and software development, sample extraction workflow simplification, strategies towards fully automated sample extraction, and best practices in automated sample preparation for regulated bioanalysis.

  20. UPLC-MS/MS determination of ptaquiloside and pterosin B in preserved natural water

    DEFF Research Database (Denmark)

    Clauson-Kaas, Frederik; Hansen, Hans Chr. Bruun; Strobel, Bjarne W.

    2016-01-01

    in environmental water samples, employing a novel internal standard. A faster, reliable, and efficient method was developed for isolation of high purity ptaquiloside and pterosin B from plant material for use as analytical standards, with purity verified by (1)H-NMR. The chemical analysis was performed by cleanup....../L for ptaquiloside and pterosin B, respectively. The use of loganin as internal standard improved repeatability of the determination of both analytes, though it could not be employed for sample preparation. Buffering raw water samples in situ with ammonium acetate to pH ∼5.5 decisively increased sample integrity......The naturally occurring carcinogen ptaquiloside and its degradation product pterosin B are found in water leaching from bracken stands. The objective of this work is to present a new sample preservation method and a fast UPLC-MS/MS method for quantification of ptaquiloside and pterosin B...

  1. Phytochemical Analysis of Pfaffia glomerata Inflorescences by LC-ESI-MS/MS

    Directory of Open Access Journals (Sweden)

    Daniele F. Felipe

    2014-09-01

    Full Text Available Pfaffia glomerata contains high levels of β-ecdysone, which has shown a range of beneficial pharmacological effects. The present study demonstrated that inflorescences of P. glomerata contain other important bioactive compounds in addition to β-ecdysone. The identification of compounds from inflorescences using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS was performed for the first time. The eight compounds identified were β-ecdysone, flavonoid glycosides such as quercetin-3-O-glucoside, kaempferol-3-O-glucoside and kaempferol-3-O-(6-p-coumaroyl-glucoside, oleanane-type triterpenoid saponins such as ginsenoside Ro and chikusetsusaponin IV, in addition to oleanonic acid and gluconic acid. This study provided information on the phytochemicals contained in P. glomerata inflorescences revealing the potential application of this plant part as raw material for the phytotherapeutic and cosmetic industries.

  2. Analysis of macamides in samples of Maca (Lepidium meyenii) by HPLC-UV-MS/MS.

    Science.gov (United States)

    McCollom, Megan M; Villinski, Jacquelyn R; McPhail, Kerry L; Craker, Lyle E; Gafner, Stefan

    2005-01-01

    The macamides are a distinct class of secondary metabolites that have so far been found only in Lepidium meyenii Walp. (Maca). Using HPLC-UV-MS/MS, the main macamides have been identified as n-benzylhexadecanamide, n-benzyl-(9Z)-octadecenamide, n-benzyl-(9Z, 12Z)-octadecadienamide, n-benzyl-(9Z, 12Z, 15Z)-octadecatrienamide and n-benzyloctadecanamide. The identities of n-benzyl-(9Z)-octadecenamide and n-benzyl-(9Z, 12Z)-octadecadienamide were confirmed by comparison of chromatographic and spectral properties with synthetic analogues. Total macamides have been quantified by HPLC-UV in plant material from different vendors using n-benzylhexadecanamide as an external standard. The amount of macamides in the dried plant material ranged from 0.0016 to 0.0123%.

  3. UHPLC-MS/MS quantification of buprenorphine, norbuprenorphine, methadone, and glucuronide conjugates in umbilical cord plasma.

    Science.gov (United States)

    Kyle, Amy Redmond; Carmical, Jennifer; Shah, Darshan; Pryor, Jason; Brown, Stacy

    2015-10-01

    Opioid use during pregnancy can result in the newborn being physically dependent on the substance, thus experiencing drug withdrawal, termed neonatal abstinence syndrome (NAS). Buprenorphine and methadone are two drugs used to treat opioid withdrawal and are approved for use in pregnancy. Quantification of these compounds in umbilical cord plasma would help assess in utero exposure of neonates in cases of buprenorphine or methadone use during pregnancy. An LC-MS/MS method using solid-phase extraction sample preparation was developed and validated for the simultaneous quantification of methadone, buprenorphine, norbuprenorphine, and glucuronide metabolites in umbilical cord plasma. The average accuracy (percentage error) and precision (relative standard deviation) were buprenorphine and metabolite concentrations. This method was successfully applied to the analysis of cord blood from women with known buprenorphine or methadone use during pregnancy. Copyright © 2015 John Wiley & Sons, Ltd.

  4. Contribution to the characterization of Opuntia spp. juices by LC-DAD-ESI-MS/MS.

    Science.gov (United States)

    Mata, A; Ferreira, J P; Semedo, C; Serra, T; Duarte, C M M; Bronze, M R

    2016-11-01

    Opuntia spp. fruits are considered as health promoting foods due to the diversity of bioactive molecules found in these fruits. The composition in organic acids, flavonols and betalains in the Opuntia ficus-indica juice from a region of Portugal was accomplished for the first time by liquid chromatography and tandem mass spectrometry using an electrospray ionization source operating in negative and positive mode. The methodology used allowed the detection of 44 compounds, from which 32 were identified. Isorhamnetin derivatives were the dominant flavonol glycosides. A total of 9 betalains including 6 betaxanthins and 3 betacyanin were also detected in the fruit juice samples and indicaxanthin, betanin and isobetanin were the major pigments. Phenolic acid and phenylpyruvic acid derivatives were also identified. To our knowledge, it is the first time derivative compounds from piscidic acid, phenolic compounds and betalains are characterized in cactus pear juice using a single LC-DAD-ESI-MS/MS method. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. [Determination of levodropropizine and its pharmacokinetics in human plasma using LC/MS/MS].

    Science.gov (United States)

    Zhao, Li-mei; Zhao, Li; Sun, Ya-xin; Qiu, Feng; Guo, Shan-bin

    2004-12-01

    To develop a rapid and sensitive LC/MS/MS method for the analysis of levodropropizine in plasma and study the pharmacokinetics of levodropropizine in healthy Chinese volunteers. Levodropropizine and zolmitriptan (internal standard, IS) were extracted from plasma samples and chromatographed on a C18 column and detected using a tandem mass spectrometer with a TurboIon Spray ionization interface. Quantitation was performed using multiple reaction monitoring (MRM) of the transitions of the m/z 237 --> m/z 120 for levodropropizine and m/z 288 --> m/z 58 for the IS. The limit of quantification of the method for levodropropizine was 0.25 microg x L(-1). The assay was linear over the concentration range from 0.25 to 500.0 microg x L(-1) and intra- and inter-day precision over this range were levodropropizine.

  6. Genotoxicity screening using biocatalyst/DNA films and capillary LC-MS/MS.

    Science.gov (United States)

    Tarun, Maricar; Bajrami, Besnik; Rusling, James F

    2006-01-15

    Detection of DNA adducts can serve as a basis for genotoxicty screening of new chemicals and drugs. We report here a simple, sensitive procedure for this purpose using films containing DNA and a biocatalyst to mimic the metabolic action of human liver cytochrome P450s. DNA adducts formed from an in-situ-generated toxic metabolite (styrene oxide) were detected at subpicomole levels after neutral thermal hydrolysis of the DNA films and analysis with capillary liquid chromatography with on-line column preconcentration and MS/MS detection. An on-line column switching system allowed for increased sample loading volume and analyte preconcentration. This approach provides an estimate of the relative rate of DNA damage.

  7. Determination of Diphenylamine in Gunshot Residue by HPLC-MS/MS

    Directory of Open Access Journals (Sweden)

    Hongcheng Mei

    2016-01-01

    Full Text Available A high performance liquid chromatography tandem mass spectrometry/mass spectrometry (HPLC-MS/MS protocol was developed for the determination of diphenylamine (DPA. Four productions of DPA were selected for qualitative assay and the peak area of the main product ion for quantitation. By means of separation using an Agilent Extend-C18 column (CA, USA (150 mm × 4.6 mm, 5 μm with methanol-water (90:10 as the mobile phase, DPA was detected by electrospray ionization (ESI tandem mass spectrometry in positive mode. The linearity of the peak area versus concentration ranged 5-500 ng/mL, r 2 = 0.9978. The limit of detection (S/N =3 of this method was 0.3 ng/mL. This method is applicable for the determination of DPA in gunshot residue.

  8. A Validated, Rapid HPLC-ESI-MS/MS Method for the Determination of Lycopsamine.

    Science.gov (United States)

    Jedlinszki, Nikoletta; Csupor, Dezső

    2015-07-01

    The aim of the present work was to develop and validate an HPLC-MS/MS method for the determination of a major pyrrolizidine alkaloid of comfrey (lycopsamine) in aqueous samples as a basis for the development of a method for the determination of absorption of lycopsamine by human skin. A linear calibration curve was established in the range of 1.32-440 ng. The intraday precision during the 3-day validation period ranged between 0.57 and 2.48% while the interday precision was 1.70% and 1.95% for quality control samples. LOD was 0.014 ng and recovery was above 97%. The lycopsamine content of the samples stored for 9 and 25 days at 22 degrees C, 10 degrees C and -25 degrees C did not vary. These results underline the good repeatability and accuracy of our method and allow the analysis of samples with very low lycopsamine content.

  9. Use of Tricaine Methanesulfonate (MS222) for Euthanasia of Reptiles

    Science.gov (United States)

    Conroy, CJ; Papenfuss, T; Parker, J; Hahn, NE

    2009-01-01

    Tricaine methanesulfonate (MS222) injected into the intracoelomic cavity of reptiles was evaluated as a chemical euthanasia method. Three western fence lizards, 2 desert iguanas, 4 garter snakes, and 6 geckos were euthanized by intracoelomic injection of 250 to 500 mg/kg of 0.7% to 1% sodium-bicarbonate–buffered MS222 solution followed by intracoelomic injection of 0.1 to 1.0 ml unbuffered 50% (v/v) MS222 solution. A simple 2-stage protocol for euthanasia of reptiles by using MS222 is outlined. In addition, the conditions for safe use of MS222 are discussed. MS222 offers an alternative to sodium pentobarbital for euthanasia of reptiles. PMID:19245747

  10. Determination of mycotoxin exposure in Germany using an LC-MS/MS multibiomarker approach.

    Science.gov (United States)

    Gerding, Johannes; Cramer, Benedikt; Humpf, Hans-Ulrich

    2014-12-01

    In this study, the exposure of a German population (n = 101) to mycotoxins was assessed using an LC-MS/MS urinary multibiomarker approach. Food consumption of the participants was documented with a food frequency questionnaire to correlate mycotoxin exposure with individual nutritional habits. The presence of 23 urinary biomarkers including trichothecenes (deoxynivalenol (DON), DON-3-glucuronide (DON-3-GlcA), T-2 toxin, HT-2 toxin (HT-2, HT-2-toxin-4-glucuronide (HT-2-GlcA), fumonisins (fumonisin B1, fumonisin B2), aflatoxins (aflatoxin B1, aflatoxin G2, aflatoxin B2, aflatoxin M1), zearalenone and derivatives (zearalanone, α-zearalenol, β-zearalenol, zearalenone-14-O-glucuronide, zearalanone-14-O-glucuronide, α-zearalenol-14-O-glucuronide/β-zearalenol-14-O-glucuronide), ochratoxin A, ochratoxin alpha, enniatin B and dihydrocitrinone was evaluated using a validated, sensitive "dilute and shoot"-LC-MS/MS method applying Scheduled MRM(TM) technology. Six mycotoxins and urinary metabolites were detected (DON, DON-3-GlcA, zearalenone-14-O-glucuronide, T-2 toxin, enniatin B, and dihydrocitrinone) in 87% of the samples in single- or co-occurence. Only DON and DON-3-GlcA were detectable in quantifiable amounts. A provisional mean daily intake of 0.52 μg DON/kg body weight was calculated. No statistical evidence for the correlation of staple food intake and urinary biomarker concentration could be determined. The results of this study suggest a low everyday exposure of the investigated German population to mycotoxins, but reveal peak exposures above the widely accepted tolerable daily intake to DON in parts of the population. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Application of UPLC-MS/MS Method for Analyzing B-vitamins in Human Milk.

    Science.gov (United States)

    Ren, Xiang Nan; Yin, Shi An; Yang, Zhen Yu; Yang, Xiao Guang; Shao, Bing; Ren, Yi Ping; Zhang, Jing

    2015-10-01

    To determine ten B-vitamins in human milk by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The pretreated human milk samples were adequately separated and quantified within 11 min by UPLC-MS/MS with an Acquity UPLC HSS T3 column (2.1×100 mm, 1.8 µm). The mobile phase was a gradient of 2.5 mmol/L ammonium formate aqueous solution and acetonitrile at a flow rate of 0.35 mL/min. Stable isotope internal standards were used in the analysis, to correct for the method variability, including matrix and ionization effects. The homogenized human milk samples were deproteinzed using methanol, unknown contaminants were extracted with diethyl ether and hydrophobic phase was discarded. The analytes were monitored via ESI+ionization and detected in multiple reaction monitoring (MRM) with three acquisition functions. Calibration curves ranged from 0.5-160 ng/mL (thiamin, riboflavin, biotin, nicotinic acid, pyridoxine, pyridoxamine, pyridoxal), and 2.5-800 ng/mL (pantothenic acid, FAD and nicotinamide) (R2=0.990-0.999). The relative recovery ranged from 80.1% to 120.2%; accuracy was determined to be 98.3% to 108.0%. Intra-day and inter-day variation were 3.4%-19.9% and 5.9%-18.1%, respectively. The limit of quantification (LOQ) for all vitamins was between 0.25 and 3 µg/L. This method was successfully applied for simultaneous analysis of ten B-vitamins in human milk. Copyright © 2015 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  12. Characterization of the Proteome of Theobroma cacao Beans by Nano-UHPLC-ESI MS/MS.

    Science.gov (United States)

    Scollo, Emanuele; Neville, David; Oruna-Concha, M Jose; Trotin, Martine; Cramer, Rainer

    2018-02-01

    Cocoa seed storage proteins play an important role in flavour development as aroma precursors are formed from their degradation during fermentation. Major proteins in the beans of Theobroma cacao are the storage proteins belonging to the vicilin and albumin classes. Although both these classes of proteins have been extensively characterized, there is still limited information on the expression and abundance of other proteins present in cocoa beans. This work is the first attempt to characterize the whole cocoa bean proteome by nano-UHPLC-ESI MS/MS analysis using tryptic digests of cocoa bean protein extracts. The results of this analysis show that >1000 proteins could be identified using a species-specific Theobroma cacao database. The majority of the identified proteins were involved with metabolism and energy. Additionally, a significant number of the identified proteins were linked to protein synthesis and processing. Several proteins were also involved with plant response to stress conditions and defence. Albumin and vicilin storage proteins showed the highest intensity values among all detected proteins, although only seven entries were identified as storage proteins. A comparison of MS/MS data searches carried out against larger non-specific databases confirmed that using a species-specific database can increase the number of identified proteins, and at the same time reduce the number of false positives. The results of this work will be useful in developing tools that can allow the comparison of the proteomic profile of cocoa beans from different genotypes and geographic origins. Data are available via ProteomeXchange with identifier PXD005586. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. QC metrics from CPTAC raw LC-MS/MS data interpreted through multivariate statistics.

    Science.gov (United States)

    Wang, Xia; Chambers, Matthew C; Vega-Montoto, Lorenzo J; Bunk, David M; Stein, Stephen E; Tabb, David L

    2014-03-04

    Shotgun proteomics experiments integrate a complex sequence of processes, any of which can introduce variability. Quality metrics computed from LC-MS/MS data have relied upon identifying MS/MS scans, but a new mode for the QuaMeter software produces metrics that are independent of identifications. Rather than evaluating each metric independently, we have created a robust multivariate statistical toolkit that accommodates the correlation structure of these metrics and allows for hierarchical relationships among data sets. The framework enables visualization and structural assessment of variability. Study 1 for the Clinical Proteomics Technology Assessment for Cancer (CPTAC), which analyzed three replicates of two common samples at each of two time points among 23 mass spectrometers in nine laboratories, provided the data to demonstrate this framework, and CPTAC Study 5 provided data from complex lysates under Standard Operating Procedures (SOPs) to complement these findings. Identification-independent quality metrics enabled the differentiation of sites and run-times through robust principal components analysis and subsequent factor analysis. Dissimilarity metrics revealed outliers in performance, and a nested ANOVA model revealed the extent to which all metrics or individual metrics were impacted by mass spectrometer and run time. Study 5 data revealed that even when SOPs have been applied, instrument-dependent variability remains prominent, although it may be reduced, while within-site variability is reduced significantly. Finally, identification-independent quality metrics were shown to be predictive of identification sensitivity in these data sets. QuaMeter and the associated multivariate framework are available from http://fenchurch.mc.vanderbilt.edu and http://homepages.uc.edu/~wang2x7/ , respectively.

  14. Library dependent LC-MS/MS acquisition via mzAPI/Live.

    Science.gov (United States)

    Webber, James T; Askenazi, Manor; Ficarro, Scott B; Iglehart, Max A; Marto, Jarrod A

    2013-05-01

    The use of MS for characterization of small molecules, nucleotides, and proteins in model organisms as well as primary tissues and clinical samples continues to proliferate at a rapid pace. The complexity and dynamic range of target analytes in biological systems hinders comprehensive analysis and simultaneously drives improvements in instrument hardware and software. As a result, state-of-the-art commercial mass spectrometers are equipped with sophisticated embedded control systems that provide robust acquisition methods accessed through intuitive graphical interfaces. Although optimized for speed, these preconfigured scan functions are otherwise closed to end-user customization beyond simple, analytical-centric parameters supplied by the manufacturer. Here, we present an open-source framework (mzAPI/Live) that enables users to generate arbitrarily complex LC-MS(n) acquisition methods via simple Python scripting. As a powerful proof-of-concept, we demonstrate real-time assignment of tandem mass spectra through rapid query of NIST peptide libraries. This represents an unprecedented capability to make acquisition decisions based on knowledge of analyte structures determined during the run itself, thus providing a path toward biology-driven MS data acquisition for the broader community. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Identification and quantitation of new glutamic acid derivatives in soy sauce by UPLC/MS/MS.

    Science.gov (United States)

    Frerot, Eric; Chen, Ting

    2013-10-01

    Glutamic acid is an abundant amino acid that lends a characteristic umami taste to foods. In fermented foods, glutamic acid can be found as a free amino acid formed by proteolysis or as a non-proteolytic derivative formed by microorganisms. The aim of the present study was to identify different structures of glutamic acid derivatives in a typical fermented protein-based food product, soy sauce. An acidic fraction was prepared with anion-exchange solid-phase extraction (SPE) and analyzed by UPLC/MS/MS and UPLC/TOF-MS. α-Glutamyl, γ-glutamyl, and pyroglutamyl dipeptides, as well as lactoyl amino acids, were identified in the acidic fraction of soy sauce. They were chemically synthesized for confirmation of their occurrence and quantified in the selected reaction monitoring (SRM) mode. Pyroglutamyl dipeptides accounted for 770 mg/kg of soy sauce, followed by lactoyl amino acids (135 mg/kg) and γ-glutamyl dipeptides (70 mg/kg). In addition, N-succinoylglutamic acid was identified for the first time in food as a minor compound in soy sauce (5 mg/kg). Copyright © 2013 Verlag Helvetica Chimica Acta AG, Zürich.

  16. A novel study of screening and confirmation of modafinil, adrafinil and their metabolite modafinilic acid under EI-GC-MS and ESI-LC-MS-MS ionization.

    Science.gov (United States)

    Dubey, S; Ahi, S; Reddy, I M; Kaur, T; Beotra, A; Jain, S

    2009-12-01

    Adrafinil and modafinil have received wide publicity and have become controversial in the sporting world when several athletes were discovered allegedly using these drugs as doping agents. By acknowledging the facts, the World Anti-Doping Agency (WADA) banned these drugs in sports since 2004. The present study explores the possibility of differentiating adrafinil and modafinil and their major metabolites under electron impact ionization in gas chromatograph-mass spectrometer (GC-MSD) and electrospray ionization in liquid chromatograph-mass spectrometer (LC-MS/MS) by studying the fragmentation pattern of these drugs. Adrafinil, modafinil and their major metabolite, modafinilic acid were analyzed on EI-GC-MSD and ESI-LC-MS/MS using various individual parameters on both the instruments. The analytical technique and equipment used in the analysis were an Agilent 6890N GC with 5973 mass selective detector for the GC-MSD analysis and an Agilent 1100 HPLC with API-3200 Triple quadrupole mass spectrometer for the LC-MS/MS analysis. Validation of both methods was performed using six replicates at different concentrations. The results show that adrafinil, modafinil and their major metabolite modafinilic acid could be detected as a single artifact without differentiation under EI-GC-MSD analysis. However, all drugs could be detected and differentiated under ESI-LCMS/MS analysis without any artifaction. The GC-MSD analysis gives a single artifact for both the drugs without differentiation and thus can be used as a marker for screening purposes. Further, the Multiple Reaction Monitoring (MRM) method developed under LC-MS/MS is fit for the purpose for confirmation of suspicious samples in routine sports testing and in forensic and clinical analysis.

  17. A new type of GC-MS with advanced capabilities

    Science.gov (United States)

    Fialkov, Alexander B.; Steiner, Urs; Jones, Larry; Amirav, Aviv

    2006-03-01

    We have combined the benefits of supersonic molecular beam interface and its related fly-through electron ionization (EI) ion source with the advanced features of the Varian 1200L gas chromatography-mass spectrometry (GC-MS) and mass spectrometry-mass spectrometry (MS-MS), resulting in a new and powerful GC-MS platform with record setting performance. Electron ionization of vibrationally cold molecules in the supersonic molecular beams (SMB) (cold EI) provided mass spectra with enhanced molecular ion, yet with good library search results and superior identification probabilities. We found that high GC column flow rates lower the elution temperature for any given compounds. This allows much larger molecules to elute at the maximum temperature of standard columns. We analyzed a mixture of heavy linear chain hydrocarbons all the way to C84H170 with a molecular weight of 1179.3 amu, using a 4 m 0.25 mm i.d. column and 32 ml/min He flow rate. Furthermore, we obtained a dominant molecular ion to all these compounds. The lower elution temperatures also greatly enhance the ability to analyze very thermally labile compounds such as carbamate pesticides. The experimental 1200 system is capable of triple quadrupole based MS-MS. We found that MS-MS on the molecular ion is much more effective than on fragment ions, and thus, the enhancement of the molecular ion directly improves the MS-MS sensitivity. Fast GC-MS analysis was also explored, based on very high column flow rate for fast splitless injections without affecting the sensitivity, and on the high system selectivity due to the combination of enhanced molecular ion and MS-MS. We demonstrate a few seconds long GC-MS-MS analysis of diazinon, spiked at 10 ng/g in a mixed fruit and vegetable extract. The feature of enhanced molecular ion provides significant enhancement in the detection sensitivity via SIM and RSIM on the molecular ion. While octafluoronaphthalene (OFN) detection limit of below 1 fg in SIM mode is shown, the

  18. Maryland magnetic fusion research program: MS speromak

    International Nuclear Information System (INIS)

    DeSilva, A.W.; Goldenbaum, G.C.; Griem, H.R.

    1989-07-01

    The main theme of our present experimentation on MS is to prolong the spheromak lifetime. This research has been concerned with such topics as passive MHD stabilization coils, impurity control and increased energy storage. At the present time the longest lived plasmas appear to be line tied to the liner or reversal coils. The natural consequence of having net flux outside the separatrix and a resistive plasma is that the plasma shrinks in time. At some point in time the plasma is far enough from the liner, or stabilization coils, that it becomes unstable. If we increase the bias field so as to move the separatrix further inside the liner, the plasma becomes unstable earlier as the separatrix moves to a smaller radius in a shorter time than if it starts out outside the liner. We have tried to circumvent this behavior with various configurations of passive conductors used as stabilizing elements. In this paper, we detail some of the machine modifications that have been tried in attempts to produce a stable, long-lived plasma

  19. LC-ESI-MS/MS identification of polar lipids of two thermophilic Anoxybacillus bacteria containing a unique lipid pattern.

    Science.gov (United States)

    Rezanka, Tomáš; Kambourova, Margarita; Derekova, Anna; Kolouchová, Irena; Sigler, Karel

    2012-07-01

    Phospholipids and glycolipids from two recently described species belonging to the thermophilic genus Anoxybacillus were analyzed by liquid chromatography-electrospray tandem mass spectrometry (LC/ESI-MS/MS). Analysis of total lipids from the facultatively anaerobic A. bogrovensis on a HILIC (Hydrophilic Interaction LIquid Chromatography) column succeeded in separating diacyl- and plasmalogen phospholipids. The LC/ESI-MS/MS analysis of the strict aerobe A. rupiensis revealed the presence of different unique polar lipids, predominantly alanyl-, lysyl-, and glucosyl-phosphatidylglycerols and cardiolipins. Each of the classes of polar lipids was then analyzed by means of the ESI-MS/MS and more than 140 molecular species of six lipid classes from A. bogrovensis and nearly 200 molecular species of nine classes of polar lipids from A. rupiensis were identified. Five classes of unidentified polar lipids were detected in both strains. Plasmalogens were thus determined for the first time in a facultatively anaerobic bacterium, i.e. A. bogrovensis.

  20. Improved LC-MS/MS of Heparan Sulfate Oligosaccharides via Chip-based Pulsed Make-up Flow

    OpenAIRE

    Huang, Yu; Shi, Xiaofeng; Yu, Xiang; Leymarie, Nancy; Staples, Gregory O.; Yin, Hongfeng; Killeen, Kevin; Zaia, Joseph

    2011-01-01

    Microfluidic chip-based hydrophilic interaction chromatography (HILIC) is a useful separation system for liquid chromatography-mass spectrometry (LC-MS) in compositional profiling of heparan sulfate (HS) oligosaccharides; however, ions observed using HILIC LC-MS are low in charge. Tandem MS of HS oligosaccharide ions with low charge results in undesirable losses of SO3 from precursor ions during collision induced dissociation. One solution is to add metal cations to stabilize sulfate groups. ...

  1. Analysis of a Brazilian green propolis from Baccharis dracunculifolia by HPLC-APCI-MS and GC-MS

    OpenAIRE

    Chang, Roberto; Piló-Veloso, Dorila; Morais, Sérgio A. L.; Nascimento, Evandro A.

    2008-01-01

    Ethanol and dichloromethane extracts of a Brazilian green propolis from Baccharis dracunculifolia were analyzed by HPLC-APCI-MS and GC-MS, respectively. The HPLC-APCI-MS technique, at the positive mode, furnished a complete and unequivocal chemical composition of the green propolis sample. It serves as fingerprint for different propolis samples. The composition of the ethanol extract consisted mainly of cinnamic acid and derivatives, flavonoids, benzoic acid and a few benzoates, non-hydroxyla...

  2. The spectral karyotype of L5178Y TK⁺/⁻ mouse lymphoma cells clone 3.7.2C and factors affecting mutant frequency at the thymidine kinase (tk) locus in the microtitre mouse lymphoma assay.

    Science.gov (United States)

    Fellows, Mick D; McDermott, Angela; Clare, Katie R; Doherty, Ann; Aardema, Marilyn J

    2014-01-01

    There has been much discussion on acceptable spontaneous mutant frequencies in the mouse lymphoma assay (MLA). This culminated in the International Workshop on Genotoxicity Testing (IWGT) recommended control limits for the microtitre version of 50-170 mutants/10(6) viable cells, which has now been included in the draft Organization for Economic Co-Operation and Development guideline for assays investigating mammalian cell gene mutation at the tk locus. Some of the factors affecting mutant frequency have been investigated. It was shown that when culturing methotrexate cleansed TK⁺/⁻ cells, a spontaneous mutant frequency of ∼100 mutants/10⁶ viable cells was achieved after only 26 doublings. However, after further culturing for ∼6 months the spontaneous mutant frequency only gradually increased. Culturing for this time did not affect the karyotype of the cell in so much as the modal chromosome number remained stable. The spontaneous mutant frequency could effectively be manipulated by cleansing with various concentrations of methotrexate. The necessity for using appropriately heat-inactivated horse serum was confirmed. Finally, following treatment with 4-nitroquinoline-N-oxide, cells did not preferentially survive when plated at high cell densities (1.6 cells plus 2,000 feeder cells/well) versus cells at low density (1.6 cells/well). It was considered that these findings confirm that the dynamics of spontaneous mutant formation in the MLA are complex. However, the karyotype of L5178Y cells is remarkably stable and assuming investigators are using cells with appropriate provenance and good culturing technique, it is clear that the IWGT recommendations are achievable. Copyright © 2013 Wiley Periodicals, Inc.

  3. MS1 Peptide Ion Intensity Chromatograms in MS2 (SWATH) Data Independent Acquisitions. Improving Post Acquisition Analysis of Proteomic Experiments*

    Science.gov (United States)

    Rardin, Matthew J.; Schilling, Birgit; Cheng, Lin-Yang; MacLean, Brendan X.; Sorensen, Dylan J.; Sahu, Alexandria K.; MacCoss, Michael J.; Vitek, Olga; Gibson, Bradford W.

    2015-01-01

    Quantitative analysis of discovery-based proteomic workflows now relies on high-throughput large-scale methods for identification and quantitation of proteins and post-translational modifications. Advancements in label-free quantitative techniques, using either data-dependent or data-independent mass spectrometric acquisitions, have coincided with improved instrumentation featuring greater precision, increased mass accuracy, and faster scan speeds. We recently reported on a new quantitative method called MS1 Filtering (Schilling et al. (2012) Mol. Cell. Proteomics 11, 202–214) for processing data-independent MS1 ion intensity chromatograms from peptide analytes using the Skyline software platform. In contrast, data-independent acquisitions from MS2 scans, or SWATH, can quantify all fragment ion intensities when reference spectra are available. As each SWATH acquisition cycle typically contains an MS1 scan, these two independent label-free quantitative approaches can be acquired in a single experiment. Here, we have expanded the capability of Skyline to extract both MS1 and MS2 ion intensity chromatograms from a single SWATH data-independent acquisition in an Integrated Dual Scan Analysis approach. The performance of both MS1 and MS2 data was examined in simple and complex samples using standard concentration curves. Cases of interferences in MS1 and MS2 ion intensity data were assessed, as were the differentiation and quantitation of phosphopeptide isomers in MS2 scan data. In addition, we demonstrated an approach for optimization of SWATH m/z window sizes to reduce interferences using MS1 scans as a guide. Finally, a correlation analysis was performed on both MS1 and MS2 ion intensity data obtained from SWATH acquisitions on a complex mixture using a linear model that automatically removes signals containing interferences. This work demonstrates the practical advantages of properly acquiring and processing MS1 precursor data in addition to MS2 fragment ion

  4. MS1 Peptide Ion Intensity Chromatograms in MS2 (SWATH) Data Independent Acquisitions. Improving Post Acquisition Analysis of Proteomic Experiments.

    Science.gov (United States)

    Rardin, Matthew J; Schilling, Birgit; Cheng, Lin-Yang; MacLean, Brendan X; Sorensen, Dylan J; Sahu, Alexandria K; MacCoss, Michael J; Vitek, Olga; Gibson, Bradford W

    2015-09-01

    Quantitative analysis of discovery-based proteomic workflows now relies on high-throughput large-scale methods for identification and quantitation of proteins and post-translational modifications. Advancements in label-free quantitative techniques, using either data-dependent or data-independent mass spectrometric acquisitions, have coincided with improved instrumentation featuring greater precision, increased mass accuracy, and faster scan speeds. We recently reported on a new quantitative method called MS1 Filtering (Schilling et al. (2012) Mol. Cell. Proteomics 11, 202-214) for processing data-independent MS1 ion intensity chromatograms from peptide analytes using the Skyline software platform. In contrast, data-independent acquisitions from MS2 scans, or SWATH, can quantify all fragment ion intensities when reference spectra are available. As each SWATH acquisition cycle typically contains an MS1 scan, these two independent label-free quantitative approaches can be acquired in a single experiment. Here, we have expanded the capability of Skyline to extract both MS1 and MS2 ion intensity chromatograms from a single SWATH data-independent acquisition in an Integrated Dual Scan Analysis approach. The performance of both MS1 and MS2 data was examined in simple and complex samples using standard concentration curves. Cases of interferences in MS1 and MS2 ion intensity data were assessed, as were the differentiation and quantitation of phosphopeptide isomers in MS2 scan data. In addition, we demonstrated an approach for optimization of SWATH m/z window sizes to reduce interferences using MS1 scans as a guide. Finally, a correlation analysis was performed on both MS1 and MS2 ion intensity data obtained from SWATH acquisitions on a complex mixture using a linear model that automatically removes signals containing interferences. This work demonstrates the practical advantages of properly acquiring and processing MS1 precursor data in addition to MS2 fragment ion

  5. Sensitive and Rapid UHPLC-MS/MS for the Analysis of Tomato Phenolics in Human Biological Samples

    Directory of Open Access Journals (Sweden)

    Miriam Martínez-Huélamo

    2015-11-01

    Full Text Available An UHPLC-MS/MS method for the quantification of tomato phenolic metabolites in human fluids was optimized and validated, and then applied in a pilot dietary intervention study with healthy volunteers. A 5-fold gain in speed (3.5 min of total run; 7-fold increase in MS sensitivity and 2-fold greater efficiency (50% peak width reduction were observed when comparing the proposed method with the reference-quality HPLC-MS/MS system, whose assay performance has been previously documented. The UHPLC-MS/MS method led to an overall improvement in the limits of detection (LOD and quantification (LOQ for all the phenolic compounds studied. The recoveries ranged between 68% and 100% in urine and 61% and 100% in plasma. The accuracy; intra- and interday precision; and stability met with the acceptance criteria of the AOAC International norms. Due to the improvements in the analytical method; the total phenolic metabolites detected in plasma and urine in the pilot intervention study were 3 times higher than those detected by HPLC-MS/MS. Comparing with traditional methods; which require longer time of analysis; the methodology described is suitable for the analysis of phenolic compounds in a large number of plasma and urine samples in a reduced time frame.

  6. Simultaneous screening of targeted and non-targeted contaminants using an LC-QTOF-MS system and automated MS/MS library searching.

    Science.gov (United States)

    Herrera-Lopez, S; Hernando, M D; García-Calvo, E; Fernández-Alba, A R; Ulaszewska, M M

    2014-09-01

    Simultaneous high-resolution full-scan and tandem mass spectrometry (MS/MS) analysis using time of flight mass spectrometry brings an answer for increasing demand of retrospective and non-targeted data analysis. Such analysis combined with spectral library searching is a promising tool for targeted and untargeted screening of small molecules. Despite considerable extension of the panel of compounds of tandem mass spectral libraries, the heterogeneity of spectral data poses a major challenge against the effective usage of spectral libraries. Performance evaluation of available LC-MS/MS libraries will significantly increase credibility in the search results. The present work was aimed to evaluate fluctuation of MS/MS pattern, in the peak intensities distribution together with mass accuracy measurements, and in consequence, performance compliant with ion ratio and mass error criteria as principles in identification processes for targeted and untargeted contaminants at trace levels. Matrix effect and ultra-trace levels of concentration (from 50 ng l(-1) to 1000 ng l(-1) were evaluated as potential source of inaccuracy in the performance of spectral matching. Matrix-matched samples and real samples were screened for proof of applicability. By manual review of data and application of ion ratio and ppm error criteria, false negatives were obtained; this number diminished when in-house library was used, while with on-line MS/MS databases 100% of positive samples were found. In our experience, intensity of peaks across spectra was highly correlated to the concentration effect and matrix complexity. In turn, analysis of spectra acquired at trace concentrations and in different matrices results in better performance in providing correct and reliable identification. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Investigation of Imposter Perfumes Using GC-MS

    Science.gov (United States)

    Mowery, Kelly A.; Blanchard, Daniel E.; Smith, Stephanie; Betts, Thomas A.

    2004-01-01

    An experiment to initiate students into several features of gas chromatograph-mass spectrometers (GC-MS) through the study of branded and counterfeit perfumes is described. The experiment enables the students to appreciate the power of GC-MS in distinguishing between original and imitation perfumes, and in analyzing data with its application…

  8. The identification of anaerobic bacteria using MALDI-TOF MS

    NARCIS (Netherlands)

    Veloo, A. C. M.; Welling, G. W.; Degener, J. E.

    Matrix Assisted Laser Desorption and Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has gained more and more popularity for the identification of bacteria. Several studies show that bacterial diagnosticis is being revolutionized by the application of MALDI-TOF MS. For anaerobic bacteria,

  9. Long-term effects of childbirth in MS

    NARCIS (Netherlands)

    D'hooghe, M.B.; Nagels, G.; Uitdehaag, B.M.J.

    2010-01-01

    Background: The uncertainty about long-term effects of childbirth presents MS patients with dilemmas. Methods: Based on clinical data of 330 female MS patients, the long-term effects of childbirth were analysed, using a cross-sectional study design. Four groups of patients were distinguished: (1)

  10. UPLC-MS/MS determination of florfenicol and florfenicol amine antimicrobial residues in tilapia muscle.

    Science.gov (United States)

    Orlando, Eduardo Adilson; Costa Roque, Aline Gabriela; Losekann, Marcos Eliseu; Colnaghi Simionato, Ana Valéria

    2016-11-01

    Despite the benefits to fish farmers, the use of antimicrobials in aquaculture has concerned consumers and competent authorities. The indiscriminate use of such substances promotes the emergence of resistant microorganisms, decreases the effectiveness of treatments, and causes possible toxic effects in humans. In Brazil, florfenicol is the only antimicrobial registered for use in aquaculture and is often used in tilapia in cage creation. Thus, this study aimed to develop a method for determination of florfenicol residues and its metabolite florfenicol amine in tilapia fillet by UPLC-MS/MS. Analytes were extracted with ethyl acetate, followed by liquid-liquid partition clean-up with hexane and SPE. The sorbents C18, phenyl and HLB-Oasis were evaluated by SPE. Phenyl sorbent showed the best results, and the extraction conditions were optimized in the sample matrix with fractional factorial design 2 4-1 . The analytes were separated on a C18 chromatographic column (50×2.1mm×1.7μm) using water (A) and acetonitrile (B) as mobile phase at a flow rate of 0.3mLmin -1 with a linear gradient (in% B): 0-2.0min: 20%; 2.0-2.5min: increase to 90%; 2.5-3.5min: 90%; 3.0-3.5min: decrease to 20%; 4.0-5.0min: 20%. The analytes were monitored in a MS/MS triple quadrupole system by MRM mode with transitions at m/z 356.1>336.1 (florfenicol) and m/z 248.1>130.1 (florfenicol amine). The optimized method was validated obtaining LOQ values of 3 and 25ngg -1 for florfenicol and florfenicol amine, respectively, precision between 20 and 36%, absolute extraction efficiency between 38 and 80%, and adequate linearity. The method was applied to samples intended for human consumption, and within the 15 evaluated samples, only one showed florfenicol residue at 30ngg -1 , which is below the maximum residue limit established in Brazil. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Organization of GC/MS and LC/MS metabolomics data into chemical libraries

    Directory of Open Access Journals (Sweden)

    DeHaven Corey D

    2010-10-01

    Full Text Available Abstract Background Metabolomics experiments involve generating and comparing small molecule (metabolite profiles from complex mixture samples to identify those metabolites that are modulated in altered states (e.g., disease, drug treatment, toxin exposure. One non-targeted metabolomics approach attempts to identify and interrogate all small molecules in a sample using GC or LC separation followed by MS or MSn detection. Analysis of the resulting large, multifaceted data sets to rapidly and accurately identify the metabolites is a challenging task that relies on the availability of chemical libraries of metabolite spectral signatures. A method for analyzing spectrometry data to identify and Quantify Individual Components in a Sample, (QUICS, enables generation of chemical library entries from known standards and, importantly, from unknown metabolites present in experimental samples but without a corresponding library entry. This method accounts for all ions in a sample spectrum, performs library matches, and allows review of the data to quality check library entries. The QUICS method identifies ions related to any given metabolite by correlating ion data across the complete set of experimental samples, thus revealing subtle spectral trends that may not be evident when viewing individual samples and are likely to be indicative of the presence of one or more otherwise obscured metabolites. Results LC-MS/MS or GC-MS data from 33 liver samples were analyzed simultaneously which exploited the inherent biological diversity of the samples and the largely non-covariant chemical nature of the metabolites when viewed over multiple samples. Ions were partitioned by both retention time (RT and covariance which grouped ions from a single common underlying metabolite. This approach benefitted from using mass, time and intensity data in aggregate over the entire sample set to reject outliers and noise thereby producing higher quality chemical identities. The

  12. Determination of Scopolamine in Human Saliva Using Solid Phase Extraction and LC/MS/MS

    Science.gov (United States)

    Wang, Zuwei; Vaksman, Zalman; Boyd, Jason; Putcha, Lakshmi

    2007-01-01

    Purpose: Scopolamine is the preferred treatment for motion sickness during space flight because of its quick onset of action, short half-life and favorable side-effect profile. The dose administered depends on the mode of administration and usually ranges between 0.1 and 0.8 mg. Such small doses make it difficult to detect concentrations of scopolamine in biological fluids by using conventional HPLC methods. To measure scopolamine in saliva and thereby to evaluate the pharmacokinetics of scopolamine, we developed an LC/MS/MS method using off-line solid phase extraction. Method: Samples (0.5mL) were loaded onto Waters Oasis HLB co-polymer cartridges (10 mg, 1 mL) and eluted with 0.5 mL methanol without evaporation and reconstitution. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 4 minutes. The mobile phase for separation was 90:10 (v/v) methanol: ammonium acetate (2 mM) in water, pH 5.0 +/- 0.1. Concentrations of scopolamine were determined using a Micromass Quattro Micro(TM) mass spectrometer with electrospray ionization (ESI). ESI mass spectra were acquired in positive ion mode with multiple reaction monitoring for the determination of scopolamine m/z = 304.2 yields 138.1 and internal standard (IS) hyoscyamine m/z = 290.2 yields 124.1. Results: The method is rapid, reproducible, specific and has the following parameters: scopolamine and the IS are eluted at 1.7 and 3.2 min respectively. The linear range is 50-5000 pg/mL for scopolamine in saliva with correlation coefficients > 0.99 with a CV < 0.5 %. The intra-day and inter-day CVs are < 15 % for quality control samples with concentrations of 75, 300, 750 and 3000 pg/mL of scopolamine in human saliva. Conclusion: Solid phase extraction allows more rapid sample preparation and greater precision than liquid extraction. Furthermore, we increased the sensitivity and specificity by adjusting the LC mobile phase and using an MS/MS

  13. Quantitative performance of a quadrupole-orbitrap-MS in targeted LC-MS determinations of small molecules.

    Science.gov (United States)

    Grund, Baptiste; Marvin, Laure; Rochat, Bertrand

    2016-05-30

    High-resolution mass spectrometry (HRMS) has been associated with qualitative and research analysis and QQQ-MS with quantitative and routine analysis. This view is now challenged and for this reason, we have evaluated the quantitative LC-MS performance of a new high-resolution mass spectrometer (HRMS), a Q-orbitrap-MS, and compared the results obtained with a recent triple-quadrupole MS (QQQ-MS). High-resolution full-scan (HR-FS) and MS/MS acquisitions have been tested with real plasma extracts or pure standards. Limits of detection, dynamic range, mass accuracy and false positive or false negative detections have been determined or investigated with protease inhibitors, tyrosine kinase inhibitors, steroids and metanephrines. Our quantitative results show that today's available HRMS are reliable and sensitive quantitative instruments and comparable to QQQ-MS quantitative performance. Taking into account their versatility, user-friendliness and robustness, we believe that HRMS should be seen more and more as key instruments in quantitative LC-MS analyses. In this scenario, most targeted LC-HRMS analyses should be performed by HR-FS recording virtually "all" ions. In addition to absolute quantifications, HR-FS will allow the relative quantifications of hundreds of metabolites in plasma revealing individual's metabolome and exposome. This phenotyping of known metabolites should promote HRMS in clinical environment. A few other LC-HRMS analyses should be performed in single-ion-monitoring or MS/MS mode when increased sensitivity and/or detection selectivity will be necessary. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Coumarin tags for analysis of peptides by MALDI-TOF MS and MS/MS. 2. Alexa Fluor 350 tag for increased peptide and protein Identification by LC-MALDI-TOF/TOF MS.

    Science.gov (United States)

    Pashkova, Anna; Chen, Hsuan-Shen; Rejtar, Tomas; Zang, Xin; Giese, Roger; Andreev, Victor; Moskovets, Eugene; Karger, Barry L

    2005-04-01

    The goal of this study was the development of N-terminal tags to improve peptide identification using high-throughput MALDI-TOF/TOF MS. Part 1 of the study was focused on the influence of derivatization on the intensities of MALDI-TOF MS signals of peptides. In part 2, various derivatization approaches for the improvement of peptide fragmentation efficiency in MALDI-TOF/TOF MS are explored. We demonstrate that permanent cation tags, while significantly improving signal intensity in the MS mode, lead to severe suppression of MS/MS fragmentation, making these tags unsuitable for high-throughput MALDI-TOF/TOF MS analysis. In the present work, it was found that labeling with Alexa Fluor 350, a coumarin tag containing a sulfo group, along with guanidation of epsilon-amino groups of Lys, could enhance unimolecular fragmentation of peptides with the formation of a high-intensity y-ion series, while the peptide intensities in the MS mode were not severely affected. LC-MALDI-TOF/TOF MS analysis of tryptic peptides from the SCX fractions of an E. coli lysate revealed improved peptide scores, a doubling of the total number of peptides, and a 30% increase in the number of proteins identified, as a result of labeling. Furthermore, by combining the data from native and labeled samples, confidence in correct identification was increased, as many proteins were identified by different peptides in the native and labeled data sets. Additionally, derivatization was found not to impair chromatographic behavior of peptides. All these factors suggest that labeling with Alexa Fluor 350 is a promising approach to the high-throughput LC-MALDI-TOF/TOF MS analysis of proteomic samples.

  15. Profiling the Oxylipin and Endocannabinoid Metabolome by UPLC-ESI-MS/MS in Human Plasma to Monitor Postprandial Inflammation.

    Science.gov (United States)

    Gouveia-Figueira, Sandra; Späth, Jana; Zivkovic, Angela M; Nording, Malin L

    2015-01-01

    Bioactive lipids, including oxylipins, endocannabinoids, and related compounds may function as specific biochemical markers of certain aspects of inflammation. However, the postprandial responsiveness of these compounds is largely unknown; therefore, changes in the circulating oxylipin and endocannabinoid metabolome in response to a challenge meal were investigated at six occasions in a subject who freely modified her usual diet. The dietary change, and especially the challenge meal itself, represented a modification of precursor fatty acid status, with expectedly subtle effects on bioactive lipid levels. To detect even the slightest alteration, highly sensitive ultra-performance liquid chromatography (UPLC) coupled to electrospray ionization (ESI) tandem mass spectrometry (MS/MS) methods for bioactive lipid profiling was employed. A previously validated UPLC-ESI-MS/MS method for profiling the endocannabinoid metabolome was used, while validation of an UPLC-ESI-MS/MS method for oxylipin analysis was performed with acceptable outcomes for a majority of the parameters according to the US Food and Drug Administration guidelines for linearity (0.9938 oxylipin and endocannabinoid metabolome. To summarize, we have shown proof-of-concept of our UPLC-ESI-MS/MS bioactive lipid protocols for the purpose of monitoring subtle shifts, and thereby useful to address lipid-mediated postprandial inflammation.

  16. Male Sex Is Independently Associated with Faster Disability Accumulation in Relapse-Onset MS but Not in Primary Progressive MS

    Science.gov (United States)

    Ribbons, Karen Ann; McElduff, Patrick; Boz, Cavit; Trojano, Maria; Izquierdo, Guillermo; Duquette, Pierre; Girard, Marc; Grand’Maison, Francois; Hupperts, Raymond; Grammond, Pierre; Oreja-Guevara, Celia; Petersen, Thor; Bergamaschi, Roberto; Giuliani, Giorgio; Barnett, Michael; van Pesch, Vincent; Amato, Maria-Pia; Iuliano, Gerardo; Fiol, Marcela; Slee, Mark; Verheul, Freek; Cristiano, Edgardo; Fernandez-Bolanos, Ricardo; Saladino, Maria-Laura; Rio, Maria Edite; Cabrera-Gomez, Jose; Butzkueven, Helmut; van Munster, Erik; Den Braber-Moerland, Leontien; La Spitaleri, Daniele; Lugaresi, Alessandra; Shaygannejad, Vahid; Gray, Orla; Deri, Norma; Alroughani, Raed; Lechner-Scott, Jeannette

    2015-01-01

    Background Multiple Sclerosis is more common in women than men and females have more relapses than men. In a large international cohort we have evaluated the effect of gender on disability accumulation and disease progression to determine if male MS patients have a worse clinical outcome than females. Methods Using the MSBase Registry, data from 15,826 MS patients from 25 countries was analysed. Changes in the severity of MS (EDSS) were compared between sexes using a repeated measures analysis in generalised linear mixed models. Kaplan-Meier analysis was used to test for sex difference in the time to reach EDSS milestones 3 and 6 and the secondary progressive MS. Results In relapse onset MS patients (n = 14,453), males progressed significantly faster in their EDSS than females (0.133 vs 0.112 per year, P<0.001,). Females had a reduced risk of secondary progressive MS (HR (95% CI) = 0.77 (0.67 to 0.90) P = 0.001). In primary progressive MS (n = 1,373), there was a significant increase in EDSS over time in males and females (P<0.001) but there was no significant sex effect on the annualized rate of EDSS change. Conclusion Among registrants of MSBase, male relapse-onset patients accumulate disability faster than female patients. In contrast, the rate of disability accumulation between male and female patients with primary progressive MS is similar. PMID:26046348

  17. Male Sex Is Independently Associated with Faster Disability Accumulation in Relapse-Onset MS but Not in Primary Progressive MS.

    Directory of Open Access Journals (Sweden)

    Karen Ann Ribbons

    Full Text Available Multiple Sclerosis is more common in women than men and females have more relapses than men. In a large international cohort we have evaluated the effect of gender on disability accumulation and disease progression to determine if male MS patients have a worse clinical outcome than females.Using the MSBase Registry, data from 15,826 MS patients from 25 countries was analysed. Changes in the severity of MS (EDSS were compared between sexes using a repeated measures analysis in generalised linear mixed models. Kaplan-Meier analysis was used to test for sex difference in the time to reach EDSS milestones 3 and 6 and the secondary progressive MS.In relapse onset MS patients (n = 14,453, males progressed significantly faster in their EDSS than females (0.133 vs 0.112 per year, P<0.001,. Females had a reduced risk of secondary progressive MS (HR (95% CI = 0.77 (0.67 to 0.90 P = 0.001. In primary progressive MS (n = 1,373, there was a significant increase in EDSS over time in males and females (P<0.001 but there was no significant sex effect on the annualized rate of EDSS change.Among registrants of MSBase, male relapse-onset patients accumulate disability faster than female patients. In contrast, the rate of disability accumulation between male and female patients with primary progressive MS is similar.

  18. Characterization of tyrosinase inhibitory constituents from the aerial parts of Humulus japonicus using LC-MS/MS coupled online assay.

    Science.gov (United States)

    Yang, Hyo Hee; Oh, Kyung-Eon; Jo, Yang Hee; Ahn, Jong Hoon; Liu, Qing; Turk, Ayman; Jang, Jae Young; Hwang, Bang Yeon; Lee, Ki Yong; Lee, Mi Kyeong

    2018-01-15

    In the screening of natural products for the development as cosmetic ingredients, the EtOAc-soluble fraction of Humulus japonicus showed tyrosinase inhibitory activity. HPLC-MS/MS coupled online tyrosinase assay of EtOAc-soluble fraction of H. japonicus characterized the twenty-eight constituents including two unknown ones and their tyrosinase inhibitory activity. Fractionation of H. japonicus using various chromatographic techniques yielded thirty-eight compounds. The chemical structures of isolated compounds were identified by spectroscopic analysis. As characterized by HPLC-MS/MS analysis, we isolated twenty-four predicted compounds and further identified two unknown ones, named humulusides A (1) and B (2). Additional ten compounds were also identified by purification. Tyrosinase inhibitory activity of isolated compounds were evaluated, which was closely correlated with the results from HPLC-MS/MS coupled online tyrosinase assay. Consistent with predicted data, two major compounds, trans-N-coumaroyltyramine (14) and cis-N-coumaroyltyramine (15) showed tyrosinase inhibition with IC 50 values of 40.6 and 36.4 μM. Taken together, H. japonicus is suggested as whitening ingredient in cosmetic products. In addition, HPLC-MS/MS coupled tyrosinase assay is powerful tool for predicting active compounds with short time and limited amounts, although identification of new compounds and verification of predicted data are also needs to be demonstrated by further experiment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Matrix effects break the LC behavior rule for analytes in LC-MS/MS analysis of biological samples

    Science.gov (United States)

    High-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are generally accepted as the preferred techniques for detecting and quantitating analytes of interest in biological matrices on the basis of the rule that one chemical compound yields one LC-...

  20. LC, MSn and LC–MS/MS studies for the characterization of degradation products of amlodipine

    Directory of Open Access Journals (Sweden)

    Ravi N. Tiwari

    2015-02-01

    Full Text Available In the present study, comprehensive stress testing of amlodipine (AM was carried out according to International Conference on Harmonization (ICH Q1A(R2 guideline. AM was subjected to acidic, neutral and alkaline hydrolysis, oxidation, photolysis and thermal stress conditions. The drug showed instability in acidic and alkaline conditions, while it remained stable to neutral, oxidative, light and thermal stress. A total of nine degradation products (DPs were formed from AM, which could be separated by the developed gradient LC method on a C18 column. The products formed under various stress conditions were investigated by LC–MS/MS analysis. The previously developed LC method was suitably modified for LC–MS/MS studies by replacing phosphate buffer with ammonium acetate buffer of the same concentration (pH 5.0. A complete fragmentation pathway of the drug was first established to characterize all the degradation products using LC–MS/MS and multi-stage mass (MSn fragmentation studies. The obtained mass values were used to study elemental compositions, and the total information helped with the identification of DPs, along with its degradation pathway. Keywords: Amlodipine, LC–MS/MS, Characterization, Degradation pathway

  1. Direct, rapid quantitative analyses of BVOCs using SIFT-MS and PTR-MS obviating sample collection

    Czech Academy of Sciences Publication Activity Database

    Smith, D.; Španěl, Patrik

    2011-01-01

    Roč. 30, č. 7 (2011), s. 945-959 ISSN 0165-9936 Institutional research plan: CEZ:AV0Z40400503 Keywords : SIFT-MS * PTR-MS * Absolute quantification Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 6.273, year: 2011

  2. Simultaneous quantitation of sphingoid bases by UPLC-ESI-MS/MS with identical (13)C-encoded internal standards

    NARCIS (Netherlands)

    Mirzaian, M.; Wisse, P.; Ferraz, M. J.; Marques, A. R. A.; Gaspar, P.; Oussoren, S. V.; Kytidou, K.; Codée, J. D. C.; van der Marel, G.; Overkleeft, H. S.; Aerts, J. M.

    2017-01-01

    Free sphingoid bases (lysosphingolipids) of primary storage sphingolipids are increased in tissues and plasma of several sphingolipidoses. As shown earlier by us, sphingoid bases can be accurately quantified using UPLC-ESI-MS/MS, particularly in combination with identical (13)C-encoded internal

  3. Analysis of pesticides in fruit, vegetables and cereals using methanolic extraction and detection by LC-MS/MS

    DEFF Research Database (Denmark)

    Granby, Kit; Andersen, Jens Hinge; Christensen, Hanne Bjerre

    2004-01-01

    in Miniprep filter HPLC vials and detected by LC–MS–MS. To compensate for variations in the MS response [13C6]-carbaryl was used as internal standard and matrix-matched pesticide solutions were used as external standards for the quantification. The method has been validated for the matrices apple, avocado...

  4. Dried blood spot validation of five immunosuppressants, without hematocrit correction, on two LC-MS/MS systems

    NARCIS (Netherlands)

    Koster, Remco A; Veenhof, Herman; Botma, Rixt; Hoekstra, Alle Tjipke; Berger, Stefan P; Bakker, Stephan JL; Alffenaar, Jan-Willem C; Touw, Daan J

    2017-01-01

    AIM: Hematocrit (Ht) effects remain a challenge in dried blood spot (DBS) sampling. The aim was to develop an immunosuppressant DBS assay on two LC-MS/MS systems covering a clinically relevant Ht range without Ht correction. RESULTS: The method was partially validated for tacrolimus, sirolimus,

  5. Simultaneous Determination of Cyclosporine A, Tacrolimus, Sirolimus, and Everolimus in Whole-Blood Samples by LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Mustafa Karapirli

    2012-01-01

    Full Text Available Objectives. Cyclosporine A (CyA, tacrolimus (TRL, sirolimus (SIR, and everolimus (RAD are immunosuppressive drugs frequently used in organ transplantation. Our aim was to confirm a robust sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS method for determination of CyA, TRL, SIR, and RAD in whole-blood samples. Materials and Methods. We used an integrated online solid-phase extraction-LC-MS/MS system and atmospheric pressure ionization tandem mass spectrometry (API-MS/MS in the multiple reaction monitoring (MRM detection mode. CyA, TRL, SIR, and RAD were simultaneously analyzed in whole blood treated with precipitation reagent taken from transplant patients. Results. System performance parameters were suitable for using this method as a high-throughput technique in clinical practice. The high concentration of one analyte in the sample did not affect the concentration of other analytes. Total analytical time was 2.5 min, and retention times of all analytes were shorter than 2 minutes. Conclusion. This LC-MS/MS method can be preferable for therapeutic drug monitoring of these immunosuppressive drugs (CyA, TRL, SRL, and RAD in whole blood. Sample preparation was too short and simple in this method, and it permits robust, rapid, sensitive, selective, and simultaneous determination of these drugs.

  6. LC-MS/MS identification of the one-carbon cycle metabolites in human plasma.

    Science.gov (United States)

    Gardner, Lidia A; Desiderio, Dominic M; Groover, Chassidy J; Hartzes, Anastasia; Yates, Charles R; Zucker-Levin, Audrey R; Bloom, Leonard; Levin, Michael C

    2013-06-01

    The one-carbon cycle is composed of four major biologically important molecules: methionine (L-Met), S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), and homocysteine (Hcy). In addition to these key metabolites, there are multiple enzymes, vitamins, and cofactors that play essential roles in the cascade of the biochemical reactions that convert one metabolite into another in the cycle. Simultaneous quantitative measurement of four major metabolites can be used to detect possible aberrations in this vital cycle. Abnormalities in the one-carbon cycle might lead to hyper- or hypomethylation, homocystinemia, liver dysfunction, and accumulation of white-matter hyperintensities in the human brain. Previously published methods describe evaluation of several components of the one-carbon cycle, but none to our knowledge demonstrated simultaneous measurement of all four key molecules (L-Met, SAM, SAH, and Hcy). We describe a novel analytical method suitable for simultaneous identification and quantification of L-Met, SAM, SAH, and Hcy with LC-MS/MS. Moreover, we tested this method to identify these metabolites in human plasma collected from patients with multiple sclerosis and healthy individuals. In a pilot feasibility study, our results indicate that patients with multiple sclerosis showed abnormalities in the one-carbon cycle. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. DETERMINATION OF COCAINE AND BENZOYLECGONINE IN BIOLOGICAL MATRICES BY HPLC AND LC-MS/MS.

    Directory of Open Access Journals (Sweden)

    Zinar Pinar GUMUS

    2016-10-01

    Full Text Available Cocaine is a powerfully addictive illicit drug. Cocaine abuse and addiction continue to increase in the world. Most analytical tests for the detection of cocaine use include the analysis of the metabolite, benzoylecgonine, in the urine. Benzoylecgonine is a major urinary metabolite of cocaine. Generally, the most common biologic matrices used for the analysis of cocaine contain the urine and blood however saliva was also added as a matrix in this study. Practical, quick, reliable, precise and accurate, reproducible analytical methods have been developed and validated for cocaine and benzoylecgonine. In addition, this chromatographic techniques were used as both initial test and confirmatory. The validated chromatographic method was successfully applied to the analysis of cocaine and benzoylecgonine compounds in synthetic biological matrix. Confirmation analyses were made by LC-MS/MS to support reliability of HPLC results. As a result of this study, it can be claimed that HPLC could be a good alternative for the analyses of various biological matrices in forensic studies. Also the matrices and concentrations were determined by HPLC instrument could be used where necessary.

  8. Multifamily determination of pesticide residues in soya-based nutraceutical products by GC/MS-MS.

    Science.gov (United States)

    Páleníková, Agneša; Martínez-Domínguez, Gerardo; Arrebola, Francisco Javier; Romero-González, Roberto; Hrouzková, Svetlana; Frenich, Antonia Garrido

    2015-04-15

    An analytical method based on a modified QuEChERS extraction coupled with gas chromatography-tandem mass spectrometry (GC-MS/MS) was evaluated for the determination of 177 pesticides in soya-based nutraceutical products. The QuEChERS method was optimised and different extraction solvents and clean-up approaches were tested, obtaining the most efficient conditions with a mixture of sorbents (PSA, C18, GBC and Zr-Sep(+)). Recoveries were evaluated at 10, 50 and 100 μg/kg and ranged between 70% and 120%. Precision was expressed as relative standard deviation (RSD), and it was evaluated for more than 160 pesticides as intra and inter-day precision, with values always below 20% and 25%, respectively. Limits of detection (LODs) ranged from 0.1 to 10 μg/kg, whereas limits of quantification (LOQs) from 0.5 to 20 μg/kg. The applicability of the method was proved by analysing soya-based nutraceuticals. Two pesticides were found in these samples, malathion and pyriproxyfen, at 11.1 and 1.5 μg/kg respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Identification and Determination of Phenolics in Lamiaceae Species by UPLC-DAD-ESI-MS/MS.

    Science.gov (United States)

    Çelik, Saliha Esin; Tufan, Ayse Nur; Bekdeser, Burcu; Özyürek, Mustafa; Güçlü, Kubilay; Apak, Resat

    2017-03-01

    This study reports the phenolic profile screening of aromatic Lamiaceae species such as marjoram (Origanum majorana L.), lavender (Lavandula officinalis) and pennyroyal (Mentha pulegium L.) using a novel and validated ultra performance liquid chromatography method coupled with DAD diode array detector and tandem mass spectrometry (MS/MS) in negative mode of electrospray ionization. Identification and quantification of phenolics in these plant extracts has been realized within 12 min. This method showed good precision (percentage relative standard deviation; RSD% 0.54-2.72 for intra-day, 1.71-4.64 for inter-day), reproducibility (percentage recovery, REC% 92.0-109.0) and linearity (r = 0.9988-0.9999). Limits of detection ranged from 0.02 to 18.2 ng/mL. The extraction of plants was performed using microwave-assisted extraction technique and 60% (v/v) aqueous methanol solvent medium was selected as suitable solvent because of maximum extraction efficiency. Total antioxidant capacity, total phenolic content and free radical scavenging activity of these plant extracts were tested and the results correlated well among each other. According to the Folin assay, phenolic contents of Origanum majorana L., Mentha pulegium L. and Lavandula officinalis were calculated as 119 ± 3.4, 85.1 ± 2.8 and 57.8 ± 2.1 mg GAE/g dry matter, respectively. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. Multi-mycotoxin stable isotope dilution LC-MS/MS method for Fusarium toxins in beer.

    Science.gov (United States)

    Habler, Katharina; Gotthardt, Marina; Schüler, Jan; Rychlik, Michael

    2017-03-01

    A stable isotope dilution LC-MS/MS multi-mycotoxin method was developed for 12 different Fusarium toxins including modified mycotoxins in beer (deoxynivalenol-3-glucoside, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyl-deoxynivalenol, HT2-toxin, T2-toxin, enniatin B, B1, A1, A, beauvericin and zearalenone). As sample preparation and purification of beer a combined solid phase extraction for trichothecenes, enniatins, beauvericin and zearalenone was firstly developed. The validation of the new method gave satisfying results: intra-day and inter-day precision and recoveries were 1-5%, 2-8% and 72-117%, respectively. In total, 61 different organic and conventional beer samples from Germany and all over the world were analyzed by using the newly developed multi-mycotoxin method. In summary, deoxynivalenol, deoxynivalenol-3-glucoside, 3-acetyldeoxynivaleneol and enniatin B were quantified in rather low contents in the investigated beer samples. None of the other monitored Fusarium toxins like 15-acetyldeoxynivalenol, HT2- and T2-toxin, zearalenone, enniatin B1, A1, A or beauvericin were detectable. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. MASPECTRAS: a platform for management and analysis of proteomics LC-MS/MS data

    Directory of Open Access Journals (Sweden)

    Rader Robert

    2007-06-01

    Full Text Available Abstract Background The advancements of proteomics technologies have led to a rapid increase in the number, size and rate at which datasets are generated. Managing and extracting valuable information from such datasets requires the use of data management platforms and computational approaches. Results We have developed the MAss SPECTRometry Analysis System (MASPECTRAS, a platform for management and analysis of proteomics LC-MS/MS data. MASPECTRAS is based on the Proteome Experimental Data Repository (PEDRo relational database schema and follows the guidelines of the Proteomics Standards Initiative (PSI. Analysis modules include: 1 import and parsing of the results from the search engines SEQUEST, Mascot, Spectrum Mill, X! Tandem, and OMSSA; 2 peptide validation, 3 clustering of proteins based on Markov Clustering and multiple alignments; and 4 quantification using the Automated Statistical Analysis of Protein Abundance Ratios algorithm (ASAPRatio. The system provides customizable data retrieval and visualization tools, as well as export to PRoteomics IDEntifications public repository (PRIDE. MASPECTRAS is freely available at http://genome.tugraz.at/maspectras Conclusion Given the unique features and the flexibility due to the use of standard software technology, our platform represents significant advance and could be of great interest to the proteomics community.

  12. LC-MS/MS analysis of neonicotinoid insecticides: Residue findings in chilean honeys

    Directory of Open Access Journals (Sweden)

    Raquel Bridi

    Full Text Available ABSTRACT Neonicotinoids are a relatively new generation of insecticides that have been used for control of pests such as aphids, leafhoppers and whiteflies. This paper presents for the first time a determination of residues of four neonicotinoid insecticides (acetamiprid, thiamethoxam, thiacloprid and imidacloprid in Chilean honey using QuEChERS extraction and UHPLC-MS/MS analysis. The limits of detection and quantification found for all analytes ranging from 0.34 to 1.43 μg kg-1 and from 0.30 to 4.76 μg kg-1, respectively. The extraction using QuEChERS method provided recoveries over 79% and the precision showed coefficient of variation lower than 20%. These data are in agreement with the international criteria that recommend general recovery limits of 70 - 120%. Of the 16 samples analyzed, in three honey samples neonicotinoids pesticides were detected. These three samples were collected from the same geographical area (Rengo. Fruit and grain production characterize the province of Rengo. The analysis of the botanical origin of these honeys showed the absence of pollen grains of crops and the majority presence of pollen grains of weeds such as Medicago sativa, Galega officinalis and Brassica rapa, which could be associated with crops. Although the residue levels found were low, the results also confirm the actual occurrence of a transfer of neonicotinoid insecticides from exposed honeybees into honey.

  13. LC-MS/MS-based metabolites of Eurycoma longifolia (Tongkat Ali) in Malaysia (Perak and Pahang).

    Science.gov (United States)

    Chua, Lee Suan; Amin, Nor Amaiza Mohd; Neo, Jason Chun Hong; Lee, Ting Hun; Lee, Chew Tin; Sarmidi, Mohamad Roji; Aziz, Ramlan Abdul

    2011-12-15

    A number of three LC-MS/MS hybrid systems (QTof, TripleTof and QTrap) has been used to profile small metabolites (m/z 100-1000) and to detect the targeted metabolites such as quassinoids, alkaloids, triterpene and biphenylneolignans from the aqueous extracts of Eurycoma longifolia. The metabolite profiles of small molecules showed four significant clusters in the principle component analysis for the aqueous extracts of E. longifolia, which had been collected from different geographical terrains (Perak and Pahang) and processed at different extraction temperatures (35°C and 100°C). A small peptide of leucine (m/z 679) and a new hydroxyl methyl β-carboline propionic acid have been identified to differentiate E. longifolia extracts that prepared at 35°C and 100°C, respectively. From the targeted metabolites identification, it was found that 3,4ɛ-dihydroeurycomanone (quassinoids) and eurylene (squalene-type triterpene) could only be detected in the Pahang extract, whereas canthin-6-one-3N-oxide could only be detected in the Perak extract. Overall, quassinoids were present in the highest concentration, particularly eurycomanone and its derivatives compared to the other groups of metabolites. However, the concentration of canthin-6-one and β-carboline alkaloids was significantly increased when the roots of the plant samples were extracted at 100°C. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Determination of anabolic-androgenic steroid adulterants in counterfeit drugs by UHPLC-MS/MS.

    Science.gov (United States)

    Cho, So-Hyun; Park, Hyoung Joon; Lee, Ji Hyun; Do, Jung-Ah; Heo, Seok; Jo, Jeong Hwa; Cho, Sooyeul

    2015-01-01

    Anabolic-androgenic steroids (AASs) have been illegally used in counterfeit drugs to improve the performance of athletes. In addition, AASs have been used for cosmetic purpose by non-athletes. To determine the presence of 26 AASs, an analysis method using ultra-liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated. The validated method was applied to 19 counterfeit drugs collected from the Internet and off-line markets during 2014. Nearly 50% (9/19) of the samples contained one of these 26 AASs. In addition, the concentration ranges of the AASs ranged from 0.09 to 119,228.57 mg/kg in the suspected samples. The determined AASs primarily consisted of testosterone and testosterone 17-propionate (26%) followed by boldenone (21%). These results indicate the adulteration of over-the-counter counterfeit drugs, and the continuous monitoring of counterfeit drugs or dubious dietary supplements containing anabolic steroids is warranted. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Determination of piroxicam from rat articular tissue and plasma based on LC-MS/MS.

    Science.gov (United States)

    Kim, Han Sol; Cho, Ha Ra; Ho, Myoung Jin; Kang, Myung Joo; Choi, Yong Seok

    2016-12-01

    Osteoarthritis (OA) is the most common type of arthritis. To manage OA, in general, oral administration of non-steroidal anti-inflammatory drugs (NSAIDs) is used. Recently, the analgesic and anti-inflammatory efficacy of piroxicam (PX), a long-acting NSAID, by intra-articular (IA) administration in OA was reported, and the possibility that PX is distributed in articular tissues at a certain concentration was raised. Thus, herein, novel LC-MS/MS methods to detect PX in rat articular tissue and plasma are presented. For articular tissue, solvent extraction with acetonitrile for 12 h was employed and a protein precipitation method was used for the preparation of a plasma sample. The developed methods were validated by following the FDA guidelines, and the validated methods were successfully applied to a PK study of IA PX. The present study presents, to our knowledge, the first method of determining a drug in articular tissue. Additionally, the level of PX in articular tissue after IA PX administration was experimentally confirmed for the first time using the present methods. Therefore, the present methods provide a new direction for in vivo evaluation for IA PX formulations and contribute to the development of alternative IA PX formulations with better effects for the treatment of OA.

  16. Simultaneous determination of sweeteners in beverages by LC-MS/MS.

    Science.gov (United States)

    Sakai, Hiroaki; Yamashita, Azusa; Tamura, Masayoshi; Uyama, Atsuo; Mochizuki, Naoki

    2015-01-01

    A new method was established for the simultaneous determination of 10 sweeteners and a degradation product in beverages by using LC-MS/MS. An ACQUITY UPLC BEH C18 (2.1 × 100 mm, 1.7 μm) was used as the LC column and 0.1% each of aqueous formic acid and formic acid in acetonitrile were used as the mobile phase. A simple and rapid determination of sweeteners was possible by diluting with a solvent, and in the case of some samples containing a large amount of foreign matter, after pre-treatment by diluting with solvent and clean-up of the sample using an Oasis HLB cartridge. All the validation results were satisfactory. As the regulations and standards for sweeteners vary from country to country, a field survey of 58 beverages marketed in Japan was performed using the present method. No issues concerning the labelling or food sanitation law were found in the tested samples.

  17. IPeak: An open source tool to combine results from multiple MS/MS search engines.

    Science.gov (United States)

    Wen, Bo; Du, Chaoqin; Li, Guilin; Ghali, Fawaz; Jones, Andrew R; Käll, Lukas; Xu, Shaohang; Zhou, Ruo; Ren, Zhe; Feng, Qiang; Xu, Xun; Wang, Jun

    2015-09-01

    Liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) is an important technique for detecting peptides in proteomics studies. Here, we present an open source software tool, termed IPeak, a peptide identification pipeline that is designed to combine the Percolator post-processing algorithm and multi-search strategy to enhance the sensitivity of peptide identifications without compromising accuracy. IPeak provides a graphical user interface (GUI) as well as a command-line interface, which is implemented in JAVA and can work on all three major operating system platforms: Windows, Linux/Unix and OS X. IPeak has been designed to work with the mzIdentML standard from the Proteomics Standards Initiative (PSI) as an input and output, and also been fully integrated into the associated mzidLibrary project, providing access to the overall pipeline, as well as modules for calling Percolator on individual search engine result files. The integration thus enables IPeak (and Percolator) to be used in conjunction with any software packages implementing the mzIdentML data standard. IPeak is freely available and can be downloaded under an Apache 2.0 license at https://code.google.com/p/mzidentml-lib/. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Tryptic peptide screening for primary immunodeficiency disease by LC/MS-MS.

    Science.gov (United States)

    Kerfoot, Sandra A; Jung, Sunhee; Golob, Karin; Torgerson, Troy R; Hahn, Si Houn

    2012-08-01

    Early diagnosis of primary immunodeficiency disorders (PIDDs) is critical for maximizing patient survival and clinical outcomes. Consequently, there is significant interest in developing broad-based, high-throughput, screening approaches capable of utilizing small blood volumes to identify patients with PIDD. We developed a novel proteomic screening approach using tandem mass spectrometry to simultaneously identify specific signature peptides derived from the transmembrane protein cluster of differentiation 3 (CD3)ɛ and the intracellular proteins Wiskott-Aldrich syndrome protein (WASP) and Bruton's tyrosine kinase (BTK) as markers of three life-threatening PIDDs; severe combined immunodeficiency, Wiskott-Aldrich syndrome, and X-linked Agammaglobulinemia. Signature peptides were analyzed by LC/MS-MS in proteolytically digested lysates from cell lines and white blood cells (WBCs). The amount of each peptide was determined by the ratio of the signature peptide peak area to that of a known amount of labeled standard peptide. Peptide concentrations were normalized to actin. We show that signature peptides from CD3ɛ, WASP, and BTK were readily detected in proteolytically digested cell lysate and their absence could correctly identify PIDD patients. This proof of concept study demonstrates the applicability of this approach to screen for PIDD and raises the possibility that it could be further multiplexed to identify additional PIDDs and potentially other disorders. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Highly sensitive analysis of sterol profiles in human serum by LC-ESI-MS/MS.

    Science.gov (United States)

    Honda, Akira; Yamashita, Kouwa; Miyazaki, Hiroshi; Shirai, Mutsumi; Ikegami, Tadashi; Xu, Guorong; Numazawa, Mitsuteru; Hara, Takashi; Matsuzaki, Yasushi

    2008-09-01

    We have developed a highly sensitive and specific method for the analysis of serum sterol profiles. Sterols in 1 mul of dried serum were derivatized into picolinyl esters (3beta-picolinate) and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using the electrospray ionization (ESI) mode. In addition to cholesterol, 19 cholesterol precursors, cholestanol, campesterol, sitosterol, and sitostanol were identified simultaneously. Quantitative analyses for the picolinyl esters of 11 available sterols were performed, and detection limits were found to be less than 1 pg on-column. Reproducibilities and recoveries of 8 noncholesterol sterols were validated according to one-way layout and polynomial equation, respectively. The variances between sample preparations and between measurements by this method were calculated to be 1.6% to 8.2% and 2.5% to 16.5%, respectively. The recovery experiments were performed using 1 mul aliquots of normal human serum spiked with 1 ng to 6 ng of sterols, and recoveries of the sterols ranged from 88.1% to 102.5% with a mean recovery of 98.1%. The present method provides reliable and reproducible results for the identification and quantification of neutral sterols, especially in small volumes of blood samples, which is useful for serological diagnosis of inherited disorders in cholesterol metabolism and for noninvasive evaluation of cholesterol biosynthesis and absorption in humans.

  20. Mycotoxin Analysis of Human Urine by LC-MS/MS: A Comparative Extraction Study

    Science.gov (United States)

    Escrivá, Laura; Font, Guillermina

    2017-01-01

    The lower mycotoxin levels detected in urine make the development of sensitive and accurate analytical methods essential. Three extraction methods, namely salting-out liquid–liquid extraction (SALLE), miniQuEChERS (quick, easy, cheap, effective, rugged, and safe), and dispersive liquid–liquid microextraction (DLLME), were evaluated and compared based on analytical parameters for the quantitative LC-MS/MS measurement of 11 mycotoxins (AFB1, AFB2, AFG1, AFG2, OTA, ZEA, BEA, EN A, EN B, EN A1 and EN B1) in human urine. DLLME was selected as the most appropriate methodology, as it produced better validation results for recovery (79–113%), reproducibility (RSDs < 12%), and repeatability (RSDs < 15%) than miniQuEChERS (71–109%, RSDs <14% and <24%, respectively) and SALLE (70–108%, RSDs < 14% and < 24%, respectively). Moreover, the lowest detection (LODS) and quantitation limits (LOQS) were achieved with DLLME (LODs: 0.005–2 μg L−1, LOQs: 0.1–4 μg L−1). DLLME methodology was used for the analysis of 10 real urine samples from healthy volunteers showing the presence of ENs B, B1 and A1 at low concentrations. PMID:29048356

  1. Multimycotoxin LC-MS/MS Analysis in Tea Beverages after Dispersive Liquid-Liquid Microextraction (DLLME).

    Science.gov (United States)

    Pallarés, Noelia; Font, Guillermina; Mañes, Jordi; Ferrer, Emilia

    2017-11-29

    The aim of the present study was to develop a multimycotoxin liquid chromatography tandem mass spectrometry (LC-MS/MS) method with a dispersive liquid-liquid microextraction procedure (DLLME) for the analysis of AFs, 3aDON, 15aDON, NIV, HT-2, T-2, ZEA, OTA, ENNs, and BEA in tea beverages and to evaluate their mycotoxin contents. The proposed method was characterized in terms of linearity, limits of detection (LODs), limits of quantification (LOQs), recoveries, repeatability (intraday precision), reproducibility (interday precision), and matrix effects to check suitability. The results show LODs in the range of 0.05-10 μg/L, LOQs in the range of 0.2-33 μg/L, and recoveries in the range of 65-127% (RSD tea, red tea, green tea, and green mint tea. The results show that, of the analyzed mycotoxins, AFB2, AFG2, 15aDON, AFG1, and ENB were detected in the samples. AFB2 (14.4-32.2 μg/L) and 15aDON (60.5-61 μg/L) presented the highest levels. Green mint tea contained the highest concentration of mycotoxins. The risk assessment study shows that the population is not much exposed to mycotoxins through the consumption of tea beverages.

  2. Using SPE-LC-ESI-MS/MS Analysis to Assess Disperse Dyes in Environmental Water Samples.

    Science.gov (United States)

    Zocolo, Guilherme Julião; Pilon dos Santos, Glauco; Vendemiatti, Josiane; Vacchi, Francine Inforçato; Umbuzeiro, Gisela de Aragão; Zanoni, Maria Valnice Boldrin

    2015-09-01

    We have optimized an SPE-LC-ESI-MS/MS method and used it to monitor disperse azo dyes in environmental aquatic samples. Calibration curves constructed for nine disperse dyes-Red 1, Violet 93, Blue 373, Orange 1, Orange 3, Orange 25, Yellow 3, Yellow 7 and Red 13-in aqueous solution presented good linearity between 2.0 and 100.0 ng mL(-1). The method provided limits of detection and quantification around 2.0 and 8.0 ng L(-1), respectively. For dyes at concentrations of 25.0 ng mL(-1), the intra- and interday analyses afforded relative standard deviation lower than 6 and 13%, respectively. The recovery values obtained for each target analyte in Milli-Q water, receiving waters and treated water samples spiked with the nine studied dyes at concentrations of 8.0, 25.0 and 50.0 ng L(-1) (n = 3) gave average recoveries greater than 70%, with RSD dyes Disperse Red 1, Disperse Blue 373 and Disperse Violet 93 at concentrations ranging from 84 to 3452 ng L(-1) in the treated effluent (TE), affluent and points collected upstream and downstream of the drinking water treatment plant of a textile dye industry in Brazil. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Pharmacokinetics of pidotimod in broiler chickens by UHPLC-MS/MS after oral and intravenous administration.

    Science.gov (United States)

    Zhang, Ruili; Qiu, Mei; Zhao, Li; Cui, Liangliang; Wang, Chunyuan; Zhu, Jiajia; Hao, Zhihui

    2018-03-01

    Pidotimod is widely used in children as an immune promoter but it has not been fully evaluated in animals. The pharmacokinetics of pidotimod and its oral bioavailability have not been described in broiler chickens. We developed a simple and sensitive UHPLC-MS/MS assay for rapid determination of pidotimod levels in chicken blood. Recoveries were nearly 100% and the coefficients of accuracy and precision were minimal. Healthy broiler chickens were given 10 mg/kg pidotimod either orally or intravenously. The oral pidotimod was rapidly absorbed (time to reach maximum concentration, 1.25 h) and rapidly eliminated (the mean residence time was 3.2 h). A noncompartmental analysis of the intravenous route indicated a mean plasma clearance of 2.2 L (h kg) -1 with an estimated mean volume of distribution at steady state of 12.69 L/kg. The bioavailability of pidotimod after oral dosing was 27%. Copyright © 2017 John Wiley & Sons, Ltd.

  4. New method for quantification of gasotransmitter hydrogen sulfide in biological matrices by LC-MS/MS

    Science.gov (United States)

    Tan, Bo; Jin, Sheng; Sun, Jiping; Gu, Zhongkai; Sun, Xiaotian; Zhu, Yichun; Huo, Keke; Cao, Zonglian; Yang, Ping; Xin, Xiaoming; Liu, Xinhua; Pan, Lilong; Qiu, Furong; Jiang, Jian; Jia, Yiqun; Ye, Fuyuan; Xie, Ying; Zhu, Yi Zhun

    2017-01-01

    Hydrogen sulfide exists widely in mammalian tissues and plays a vital role in physiological and pathophysiological processes. However, striking differences with orders of magnitude were observed for the detected hydrogen sulfide concentrations in biological matrices among different measurements in literature, which lead to the uncertainty for examination the biological relevance of hydrogen sulfide. Here, we developed and validated a liquid chromatography- mass spectrometry (LC-MS/MS) method for the determination of hydrogen sulfide in various biological matrices by determination of a derivative of hydrogen sulfide and monobromobimane named sulfide dibimane (SDB). 36S-labeled SDB was synthesized and validated for using as an internal standard. This method has been successfully used to measure hydrogen sulfide levels in a broad range of biological matrices, such as blood, plasma, tissues, cells, and enzymes, across different species. Moreover, a novel mode that hydrogen sulfide could loosely and non-covalently bind to human serum protein (HSA) and hemoglobin (HB) was revealed by using the developed method. PMID:28406238

  5. Staphylococcus aureus methicillin resistance detected by HPLC-MS/MS targeted metabolic profiling.

    Science.gov (United States)

    Schelli, Katie; Rutowski, Joshua; Roubidoux, Julia; Zhu, Jiangjiang

    2017-03-15

    Recently, novel bioanalytical methods, such as NMR and mass spectrometry based metabolomics approaches, have started to show promise in providing rapid, sensitive and reproducible detection of Staphylococcus aureus antibiotic resistance. Here we performed a proof-of-concept study focused on the application of HPLC-MS/MS based targeted metabolic profiling for detecting and monitoring the bacterial metabolic profile changes in response to sub-lethal levels of methicillin exposure. One hundred seventy-seven targeted metabolites from over 20 metabolic pathways were specifically screened and one hundred and thirty metabolites from in vitro bacterial tests were confidently detected from both methicillin susceptible and methicillin resistant Staphylococcus aureus (MSSA and MRSA, respectively). The metabolic profiles can be used to distinguish the isogenic pairs of MSSA strains from MRSA strains, without or with sub-lethal levels of methicillin exposure. In addition, better separation between MSSA and MRSA strains can be achieved in the latter case using principal component analysis (PCA). Metabolite data from isogenic pairs of MSSA and MRSA strains were further compared without and with sub-lethal levels of methicillin exposure, with metabolic pathway analyses additionally performed. Both analyses suggested that the metabolic activities of MSSA strains were more susceptible to the perturbation of the sub-lethal levels of methicillin exposure compared to the MRSA strains. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. UHPLC/ESI-MS/MS Determination of 187 Pesticides in Wine.

    Science.gov (United States)

    Wang, Jian; Cheung, Wendy

    2016-01-01

    This paper presents an ultra HPLC/electrospray ionization-tandem MS method to determine pesticides in wine. We adopted the quick, easy, cheap, effective, rugged, and safe (QuEChERs) method for extraction and used core-shell column to achieve ultra-HPLC to develop and validate a simple and fast method to analyze 187 pesticide residues in red and white wine samples. Pesticide residues were extracted from wine samples using QuEChERS. Ultra HPLC/electrospray ionization-tandem MS quantification was achieved using matrix-matched standard calibration curves with isotopically labeled standards or a chemical analogue as internal standards with an analytical range from 5.0 to 500.0 μg/L. The method performance characteristics that included overall recovery, intermediate precision, and measurement uncertainty were evaluated according to a nested experimental design. Generally, 98.4% (in red wine) and 96.8% (in white wine) of the pesticides had recoveries between 71 and 120%; 98.9% (in red wine) and 99.5% (in white wine) of the pesticides had the intermediate precision ≤20%; and 99.5% (in red wine) and 98.4% (in white wine) of the pesticides had measurement uncertainty ≤50%.

  7. Quantitative Determination of Ivermectin in Raw Milk Using Positive ESI LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Meenakshi Dahiya

    2010-01-01

    Full Text Available Ivermectin, a veterinary drug, is commonly used endectocide for animal husbandry. The drug is available in the form of subcutaneous or topical formulations. Its application may cause accumulation of its residues into the animal tissues, which ultimately find their way into the food products, such as milk and meat products. In order to determine the residues of ivermectin in milk, a comparatively simple, sensitive and rapid method was developed and validated using LC-MS/MS. The MRM transitions corresponding to m/z 892.71>569.6, 892.71>551.5 and 892.71>307.3 were used for the purpose of quantification and evaluation of other parameters of the method. The limit of detection of the method was found to be 0.1 μg/kg and the limit of quantitation was calculated as 0.2 μg/kg. The method was found to be linear in the range of 1.0 ng/mL to 100.0 ng/mL with correlation coefficient of 0.9992 for pure calibration curve and 0.9990 for the matrix- matched calibration curve. The recoveries of ivermectin from the spiked samples of raw milk were found between 85 to 105%.

  8. Simultaneous quantification of seven hippocampal neurotransmitters in depression mice by LC-MS/MS.

    Science.gov (United States)

    Huang, Fei; Li, Jia; Shi, Hai-Lian; Wang, Ting-ting; Muhtar, Wahaf; Du, Min; Zhang, Bei-bei; Wu, Hui; Yang, Li; Hu, Zhi-bi; Wu, Xiao-jun

    2014-05-30

    There is no method available to simultaneously detect GABA, Glu, Epi, NE, DA, 5-HT and 5-HIAA in mouse hippocampus. A rapid and sensitive LC-MS/MS method has been developed for simultaneously measuring seven neurotransmitters in mouse hippocampus. The analytes were detected in positive mode with multiple reaction monitoring (MRM) and the procedure was completed in less than 9min. This method exhibited excellent linearity for all of the analytes with regression coefficients higher than 0.99, and showed good intra- and inter-day precisions (RSDneurotransmitters in a mouse depression model induced by successive methylprednisolone injections. The results indicated that this depression model was closely associated with the decreased level of Epi (p=0.002) and elevated ratio of 5-HIAA/5-HT (p=0.01), which has never been reported elsewhere. Compared with previous methods, current approach is more convenient without any pre-column derivatization of the analytes but enhances detectability with incremental neurotransmitter profile and shortens detection time. This work represents the first accurate simultaneous determination of seven neurotransmitters in the mouse depression model induced by methylprednisolone. The reliable method will benefit the research of neurological diseases with the altered neurotransmitter profile in brain. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. HPLC-MS/MS investigation of biochemical markers for the disclosure of erythropoietin abuse in sports

    Science.gov (United States)

    Appolonova, S. A.; Dikunets, M. A.; Rodchenkov, G. M.

    2009-04-01

    The polypeptide hormone erythropoietin (EPO), which is a forbidden doping drug, was determined by high-performance liquid chromatography combined with tandem mass spectrometry (HPLC-MS/MS). The hypothesis about the influence of EPO on the asymmetric dimethylarginine (ADMA)-dimethylargininedime-thylaminohydrolase (DDAH)-NO-synthase system was verified. Changes in this system can serve as indirect biochemical markers of the presence of the forbidden EPO drug in the organism. In the test group, the concentrations of biochemical markers varied from 10 to 40 μg/ml for ADMA and symmetrical DMA (SDMA) and from 0.5 to 10 μg/ml for arginine and citrulline. A single intravenous administration of r-HuEPO (Epocrin, 2000 ME/day) for two volunteers reliably increased ADMA, SDMA, arginine, and citrulline concentrations to 40-270 μg/ml, 40-240μg/ml, 10-60 μg/ml, and 12-140 μg/ml, respectively, with respect to the reference values. The simultaneous increase in arginine, methylarginines, and citrulline contents could be an indirect marker of EPO abuse. The method is recommended for fast screening analysis.

  10. Moxidectin residues in lamb tissues: Development and validation of analytical method by UHPLC-MS/MS.

    Science.gov (United States)

    Del Bianchi A Cruz, Michelle; Fernandes, Maria A M; de C Braga, Patricia A; Monteiro, Alda L G; Daniel, Daniela; Reyes, Felix G R

    2018-01-01

    The development and validation of a throughput method for the quantitation of moxidectin residues in lamb target tissues (muscle, kidney, liver and fat) was conducted using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). To achieve higher recovery of the analyte from the matrices, a modified QuEChERS method was used for sample preparation. The chromatographic separation was achieved using a Zorbax Eclipse Plus C18 RRHD column with a mobile phase comprising 5mM ammonium formate solution +0.1% formic acid (A) and acetonitrile +0.1% formic acid (B) in a linear gradient program. Method validation was performed based on the Commission Decision 2002/657/EC and VICH GL49. To quantify the analyte, matrix-matched analytical curves were constructed with spiked blank tissues, with a limit of quantitation of 5ngg -1 and limit of detection of 1.5ngg -1 for all matrices. The linearity, decision limit, detection capability accuracy, and inter- and intra-day repeatability of the method are reported. The method was successfully applied to incurred lamb tissue samples (muscle, liver, kidney and fat) in a concentration range from 5 to 200μgkg -1 , which demonstrated its suitability for monitoring moxidectin residues in lamb tissues in health surveillance programs, as well as for pharmacokinetics and residue depletion studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. UHPLC-MS/MS based target profiling of stress-induced phytohormones.

    Science.gov (United States)

    Floková, Kristýna; Tarkowská, Danuše; Miersch, Otto; Strnad, Miroslav; Wasternack, Claus; Novák, Ondřej

    2014-09-01

    Stress-induced changes in phytohormone metabolite profiles have rapid effects on plant metabolic activity and growth. The jasmonates (JAs) are a group of fatty acid-derived stress response regulators with roles in numerous developmental processes. To elucidate their dual regulatory effects, which overlap with those of other important defence-signalling plant hormones such as salicylic acid (SA), abscisic acid (ABA) and indole-3-acetic acid (IAA), we have developed a highly efficient single-step clean-up procedure for their enrichment from complex plant matrices that enables their sensitive quantitative analysis using hyphenated mass spectrometry technique. The rapid extraction of minute quantities of plant material (less than 20mg fresh weight, FW) into cold 10% methanol followed by one-step reversed-phase polymer-based solid phase extraction significantly reduced matrix effects and increased the recovery of labile JA analytes. This extraction and purification protocol was paired with a highly sensitive and validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method and used to simultaneously profile sixteen stress-induced phytohormones in minute plant material samples, including endogenous JA, several of its biosynthetic precursors and derivatives, as well as SA, ABA and IAA. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Surveillance of drug abuse in Hong Kong by hair analysis using LC-MS/MS.

    Science.gov (United States)

    Leung, K Wing; Wong, Zack C F; Ho, Janet Y M; Yip, Ada W S; Cheung, Jerry K H; Ho, Karen K L; Duan, Ran; Tsim, Karl W K

    2017-12-04

    The aim of this study is to reveal the habits of drug abusers in hair samples from drug rehabilitation units in Hong Kong. With the application of liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology, a total of 1771 hair samples were analyzed during the period of hair testing service (January 2012 to March 2016) provided to 14 drug rehabilitation units including non-governmental organizations (NGOs), rehabilitation centers, and medical clinics. Hair samples were analyzed for abused drugs and their metabolites simultaneously, including ketamine, norketamine, cocaine, benzoylecgonine, cocaethylene, norcocaine, codeine, MDMA, MDA, MDEA, amphetamine, methamphetamine, morphine, 6-acetylmorphine, phencyclidine, and methadone. The results showed that ketamine (77.2%), cocaine (21.3%), and methamphetamine (16.5%) were the frequently detected drugs among those drug abusers, which is consistent with the reported data. In addition, the usage of multiple drugs was also observed in the hair samples. About 29% of drug-positive samples were detected with multiple drug use. Our studies prove that our locally developed hair drug-testing method and service can be a valid tool to monitor the use of abused drugs, and which could facilitate rehabilitation program management. Copyright © 2017 John Wiley & Sons, Ltd.

  13. Determination of raloxifene hydrochloride in human urine by LC-MS-MS

    Directory of Open Access Journals (Sweden)

    K. Tharpa

    2009-09-01

    Full Text Available A sensitive and selective liquid chromatographic-tandem mass spectrometric (LC-MS-MS method was developed to determine raloxifene hydrochloride (RLX in human urine. After a solid-phase extraction with SPE cartridge, the urine sample was analyzed on a C18 column (Symmetry 3.5μm; 50 mm4.6 mm i.d interfaced with a triple quadruple tandem mass spectrometer. A positive electrospray ionization was employed as the ionization source. The mobile phase consisted of ammonium acetate (pH 4.0–acetonitrile (60:40, v/v.The method was linear over a concentration range of 20–1000 ng mL-1. The lower limit of quantitation was 20 ng mL-1. The intra-day and inter-day relative standard deviation across three validation runs over the entire concentration range was <10.5%. The accuracy determined at three concentrations (50, 500 and 850 ng mL-1 RLX was within ±0.84% in terms of relative errors.

  14. Substitution pattern elucidation of hydroxypropyl Pinus pinaster (Ait.) bark polyflavonoid derivatives by ESI(-)-MS/MS.

    Science.gov (United States)

    García Marrero, Danny E; Glasser, Wolfgang G; Pizzi, Antonio; Paczkowski, Sebastian; Laborie, Marie-Pierre G

    2014-10-01

    The structure of condensed tannins (CTs) from Pinus pinaster bark extract and their hydroxypropylated derivatives with four degrees of substitution (DS 1, 2, 3 and 4) has been characterized for the first time using negative-ion mode electrospray ionization tandem mass spectrometry (ESI(-)-MS/MS). The results showed that P. pinaster bark CTs possess structural homogeneity in terms of monomeric units (C(15), catechin). The oligomer sizes were detected to be dimers to heptamers. The derivatives showed typical phenyl-propyl ether mass fragmentation by substituent elimination (58 amu) and inherent C(15) flavonoid fissions. The relative abundance of the product ions revealed a preferential triple, tetra-/penta- and octa- hydroxypropylation substitution pattern in the monomer, dimer and trimer derivatives, respectively. A defined order of -OH reactivity towards propylene oxide was established by means of multistage experiments (A-ring ≥ B-ring > C-ring). A high structural heterogeneity of the modified oligomers was detected. Copyright © 2014 John Wiley & Sons, Ltd.

  15. LC-MS-MS quantitative analysis reveals the association between FTO and DNA methylation.

    Directory of Open Access Journals (Sweden)

    Yuting Zhu

    Full Text Available Fat mass and obesity-associated protein (FTO is α-ketoglutarate-dependent dioxygenase and responsible for demethylating N6-methyladenosine (m6A in mRNA, 3-methylthymine (m3T in single-stranded DNA (ssDNA and 3-methyluracil (m3U in single-stranded RNA (ssRNA. Its other function remains unknown but thousands of mammalian DNA show 5-methyl-2'-deoxycytidine (5mdC modification and 5mdC demethylases are required for mammalian energy homeostasis and fertility. Here, we aimed to confirm whether FTO proteins can demethylate 5mdC in DNA. However, we found that FTO exhibits no potent demethylation activity against 5mdC in vitro and in vivo by using liquid chromatography-tandem mass spectrometry (LC-MS-MS. The result showed FTO demethylase has the characteristics of high substrates specificity and selectivity. In addition, we also used immunofluorescence technique to demonstrate overexpression of wild type TET2, but not FTO and mutant TET2 in Hela cells results in higher levels of 5-hydroxymethyl-2'-deoxycytidine (5hmdC generated from 5mdC. In conclusion, our results not only reveal the enzymatic activity of FTO, but also may facilitate the future discovery of proteins involved in epigenetic modification function.

  16. ESI-MS2and Anti-inflammatory Studies of Cyclopropanic Triterpenes. UPLC-ESI-MS and MS2Search of Related Metabolites from Donella ubanguiensis.

    Science.gov (United States)

    Sandjo, Louis P; Nascimento, Marcus V P Dos Santos; da Silva, Layzon A L; Munhoz, Antonio C M; Pollo, Luiz A E; Biavatti, Maique W; Ngadjui, Bonaventure T; Opatz, Till; Fröde, Tania S

    2017-01-01

    Triterpenes are one of the largest secondary metabolites groups spread in the plant kingdom with various skeletons. These metabolites have showed various bioactivities including anti-inflammatory activity. The study aims to explore the mass spectrometry fragmentation of donellanic acids A-C (DA A-C), three compounds identified from Donella ubanguiensis; in addition, the fragmentation behaviour of these metabolites will serve as a fingerprint to search and characterise triterpenes congeners in fruits, bark and wood crude extracts of D. ubanguiensis. This work was prompted by the anti-inflammatory activity on leukocyte migration, exudate concentrations and myeloperoxidase activity obtained for DA A-B. The bioactivity was performed on mouse model of pleurisy induced by carrageenan and the parameters were analysed by veterinarian automated cell counter and colorimetric assays. While the tandem mass analyses of DA A-C were carried out by a direct infusion ESI-QTOF-MS/MS, the extracts were studied by UPLC-ESI-QTOF-MS and UPLC-ESI-QTOF-MS/MS. DA A displayed interesting anti-inflammatory activity by inhibiting leukocyte migration, exudate concentrations and myeloperoxidase activity (p  0.05). Moreover, the diagnostic of the MS 2 behaviour of DA A-C in conjunction with the chromatograms and the obtained MS 2 data of the crude extract led to the characterisation of three cyclopropane triterpenes (T1-T3) and six saponins (T4-T9) from the fruits, the bark, and the wood extracts. Donella species deserve more investigation since metabolites related to the anti-inflammatory compound (DA A) could be identified. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  17. [Functional analysis of cancer-derived immunoglobulin G whole molecule-interacting proteins identified by LC-MS/MS].

    Science.gov (United States)

    Wang, Ju-Ping; Chen, Han-Ying; Peng, Hui

    2015-01-01

    To identify cancer-derived immunoglobulin G (IgG) whole molecule-interacting proteins to provide important clues for studying IgG biological functions. HeLa cell lysate was immunoprecipitated with rabbit antihuman IgG whole molecule antibody and normal rabbit IgG. The immunocomplex underwent sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and was detected with silver staining. Three prominently enhanced bands were subjected to protein identification with liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the MS data were analyzed with Swiss-Prot database. Cancer-derived IgG whole molecule-interacting proteins were screened and functionally annotated. We identified 6 potential cancer-derived IgG whole molecule-interacting proteins with co-immunoprecipitation combined with LC-MS/MS, which provides valuable clues for studying the function of cancer-derived IgG.

  18. Quantitative analysis of pharmacokinetic study samples by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS).

    Science.gov (United States)

    Mück, W

    1999-09-01

    The basis of all pharmacokinetic evaluations are powerful assays to quantify drugs and/or metabolites in biological matrices using modern sensitive instrumental analytical techniques, such as capillary gas chromatography and high-performance liquid chromatography (HPLC). Being both specific and universal, mass spectrometry (MS) is an ideal chromatographic detector. Due to recent exciting achievements in the interfacing of liquid chromatography (LC) and MS, LC-MS, like the successfully preceding hyphenated technique gas chromatography-mass spectrometry (GC-MS), has now become a valuable technique in the analyst's toolbox. The key features of LC-MS are explained and four examples demonstrating its potential for highly specific and sensitive routine drug assays with the option of high sample throughput in pharmacokinetic investigations are presented.

  19. The potential use of complex derivatization procedures in comprehensive HPLC-MS/MS detection of anabolic steroids.

    Science.gov (United States)

    Baranov, Pavel A; Appolonova, Svetlana A; Rodchenkov, Grigory M

    2010-10-01

    The use of two separate derivatization procedures with the formation of oxime (hydroxyl ammonium pretreatment) and picolinoyl (mixed anhydride method) derivates of anabolic steroids following HPLC-MS/MS analysis was proposed. The main product ions of obtained derivatives for 21 anabolic steroids were evaluated and fragmentation pathways were compared.The analysis of MS/MS spectra for underivatized steroids versus oxime or picolinoyl derivatives showed that in case of analytes containing conjugated double bonds in sterane core all of the observed MS/MS spectra contained abundant product ions of diagnostic value. The implementation of derivatization procedures to such compounds is useful for upgrading sensitivity or selectivity of the evaluated method. On the other hand, MS/MS spectra of underivatized and oxime analytes without conjugated double bonds in sterane core produce spectra with large amounts of low abundant product ions. Picolinoyl derivatives formation leads to highly specific spectra with product ions of diagnostic value coupled with sensitive and selective analysis at the same time. The intra- and inter-group comparison analysis revealed that fragmentation pathways for underivatized steroids and correspondent oxime derivatives are similar.The obtained oxime and picolinoyl derivatives provided 10-15 times higher ESI response in the HPLC-ESI-MS-selected reaction monitoring (SRM) when compared to those of underivatized molecules in positive HPLC-ESI-MS mode.Due to the laborious sample preparation we suggest to use the performed strategy for confirmation analysis purposes, metabolic studies or while the identification of new steroids or steroid-like substances. Copyright © 2010 John Wiley & Sons, Ltd.

  20. Determination of Voriconazole Concentrations in Serum by GC-MS.

    Science.gov (United States)

    Smith, Alisha; Leung-Pineda, Van

    2016-09-01

    Voriconazole is a broad spectrum triazole antifungal drug used to treat systemic fungal infections. Therapeutic drug monitoring of voriconazole is necessary for achieving maximal efficiency without inducing toxic side effects. Other publications have reported methods for measuring voriconazole in serum using high-performance liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry (LC-MS/MS). Here, we report for the first time a method for the measurement of voriconazole in serum samples using gas chromatography mass spectrometry (GC-MS). Protein precipitation with methanol was used to extract the antifungal that was derivatized with BSTFA (Sigma-Aldrich, St. Louis, MO) and analyzed by GC-MS. Linearity, sensitivity, precision, accuracy, and drug interferences were evaluated for this assay. Our method was linear up to 10 μg/ml of voriconazole. The LOQ was determined to be 0.4 μg/ml. CV for between-day precision was 10% error in measurement. To our knowledge, we report here the first GC-MS method for voriconazole measurement with acceptable performance. We hope that this method allows clinical laboratories without HPLC or LC-MS/MS instrumentation to measure voriconazole. © 2015 Wiley Periodicals, Inc.

  1. LA-ICP-MS of magnetite: Methods and reference materials

    Science.gov (United States)

    Nadoll, P.; Koenig, A.E.

    2011-01-01

    Magnetite (Fe3O4) is a common accessory mineral in many geologic settings. Its variable geochemistry makes it a powerful petrogenetic indicator. Electron microprobe (EMPA) analyses are commonly used to examine major and minor element contents in magnetite. Laser ablation ICP-MS (LA-ICP-MS) is applicable to trace element analyses of magnetite but has not been widely employed to examine compositional variations. We tested the applicability of the NIST SRM 610, the USGS GSE-1G, and the NIST SRM 2782 reference materials (RMs) as external standards and developed a reliable method for LA-ICP-MS analysis of magnetite. LA-ICP-MS analyses were carried out on well characterized magnetite samples with a 193 nm, Excimer, ArF LA system. Although matrix-matched RMs are sometimes important for calibration and normalization of LA-ICP-MS data, we demonstrate that glass RMs can produce accurate results for LA-ICP-MS analyses of magnetite. Cross-comparison between the NIST SRM 610 and USGS GSE-1G indicates good agreement for magnetite minor and trace element data calibrated with either of these RMs. Many elements show a sufficiently good match between the LA-ICP-MS and the EMPA data; for example, Ti and V show a close to linear relationship with correlation coefficients, R2 of 0.79 and 0.85 respectively. ?? 2011 The Royal Society of Chemistry.

  2. Quantification of acetaminophen (paracetamol) in human plasma and urine by stable isotope-dilution GC-MS and GC-MS/MS as pentafluorobenzyl ether derivative.

    Science.gov (United States)

    Trettin, Arne; Zoerner, Alexander A; Böhmer, Anke; Gutzki, Frank-Mathias; Stichtenoth, Dirk O; Jordan, Jens; Tsikas, Dimitrios

    2011-08-01

    We report on the quantitative determination of acetaminophen (paracetamol; NAPAP-d(0)) in human plasma and urine by GC-MS and GC-MS/MS in the electron-capture negative-ion chemical ionization (ECNICI) mode after derivatization with pentafluorobenzyl (PFB) bromide (PFB-Br). Commercially available tetradeuterated acetaminophen (NAPAP-d(4)) was used as the internal standard. NAPAP-d(0) and NAPAP-d(4) were extracted from 100-μL aliquots of plasma and urine with 300 μL ethyl acetate (EA) by vortexing (60s). After centrifugation the EA phase was collected, the solvent was removed under a stream of nitrogen gas, and the residue was reconstituted in acetonitrile (MeCN, 100 μL). PFB-Br (10 μL, 30 vol% in MeCN) and N,N-diisopropylethylamine (10 μL) were added and the mixture was incubated for 60 min at 30 °C. Then, solvents and reagents were removed under nitrogen and the residue was taken up with 1000 μL of toluene, from which 1-μL aliquots were injected in the splitless mode. GC-MS quantification was performed by selected-ion monitoring ions due to [M-PFB](-) and [M-PFB-H](-), m/z 150 and m/z 149 for NAPAP-d(0) and m/z 154 and m/z 153 for NAPAP-d(4), respectively. GC-MS/MS quantification was performed by selected-reaction monitoring the transition m/z 150 → m/z 107 and m/z 149 → m/z 134 for NAPAP-d(0) and m/z 154 → m/z 111 and m/z 153 → m/z 138 for NAPAP-d(4). The method was validated for human plasma (range, 0-130 μM NAPAP-d(0)) and urine (range, 0-1300 μM NAPAP-d(0)). Accuracy (recovery, %) ranged between 89 and 119%, and imprecision (RSD, %) was below 19% in these matrices and ranges. A close correlation (r>0.999) was found between the concentrations measured by GC-MS and GC-MS/MS. By this method, acetaminophen can be reliably quantified in small plasma and urine sample volumes (e.g., 10 μL). The analytical performance of the method makes it especially useful in pediatrics. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Multidimensional liquid chromatography with parallel ICP MS and electrospray MS/MS detection as a tool for the characterization of arsenic species in algae

    Energy Technology Data Exchange (ETDEWEB)

    McSheehy, S.; Pohl, P.; Velez, D.; Szpunar, J. [CNRS UMR, Pau-Pyrenees (France)

    2002-02-01

    An analytical strategy was developed for the characterization of arsenic species in a Laminaria algae. The approach was based on multidimensional liquid chromatography (LC) including sample extract cleanup by size-exclusion LC, separation of arsenic species by anion-exchange LC, verification of the chromatographic purity of arsenic-containing fractions, and their further purification, if necessary, by reversed-phase (RP) HPLC. The complementarity of ICP MS, used as the chromatographic detector, and ES MS/MS, employed for the identification of the peaks observed, was demonstrated. The species found were: arsenosugar A 11.7{+-}0.5 {mu}g g{sup -1}, As{sup V} 10.9{+-}2.1 {mu}g g{sup -1}, arsenosugar B 2.22{+-}0.07 {mu}g g{sup -1}, arsenosugar D 1.5{+-}1.2 {mu}g g{sup -1}, a newly detected arsenosugar 1.13{+-}0.07 {mu}g g{sup -1}, arsenosugar C 0.61{+-}0.04 {mu}g g{sup -1}, DMA 0.42{+-}0.02 {mu}g g{sup -1} and these accounted for >99% of the arsenic present. The identities of all the species, except the newly detected compound, were doubly checked by matching the retention times of chromatographically pure (after the 3{sup rd} LC dimension) species with standards and by ES MS/MS. (orig.)

  4. New metabolites of fenofibrate in Sprague–Dawley rats by UPLC-ESI-QTOF-MS-based metabolomics coupled with LC-MS/MS

    Science.gov (United States)

    Liu, A.; Chen, Y.; Yang, Z.; Feng, Y.; Rui, W.; Luo, W.; Liu, Y.; Gonzalez, F. J.; Dai, R.

    2009-01-01

    Fenofibrate has been widely used for the treatment of dyslipidaemia with a long history. Species differences of its metabolism were reported, but its metabolites in rodent have not been fully investigated.Urine and plasma samples were collected before and after oral dosages of fenofibrate in Sprague–Dawley rats. Urine samples were subjected to ultra-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) analysis, and projection to latent structures discriminant analysis was used for the identification of metabolites.New metabolites in urine and plasma were also studied by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The metabolism pathway was studied in rat hepatocytes. Synthesized and purchased authentic compounds were used for metabolite identification by LC-MS/MS.Five ever-reported metabolites were identified and another four new ones were found. Among these new metabolites, fenofibric acid taurine and reduced fenofibric acid taurine indicate new phase II conjugation pathway of fenofibrate. PMID:19350456

  5. Precise and accurate isotope ratio measurements by ICP-MS.

    Science.gov (United States)

    Becker, J S; Dietze, H J

    2000-09-01

    The precise and accurate determination of isotope ratios by inductively coupled plasma mass spectrometry (ICP-MS) and laser ablation ICP-MS (LA-ICP-MS) is important for quite different application fields (e.g. for isotope ratio measurements of stable isotopes in nature, especially for the investigation of isotope variation in nature or age dating, for determining isotope ratios of radiogenic elements in the nuclear industry, quality assurance of fuel material, for reprocessing plants, nuclear material accounting and radioactive waste control, for tracer experiments using stable isotopes or long-lived radionuclides in biological or medical studies). Thermal ionization mass spectrometry (TIMS), which used to be the dominant analytical technique for precise isotope ratio measurements, is being increasingly replaced for isotope ratio measurements by ICP-MS due to its excellent sensitivity, precision and good accuracy. Instrumental progress in ICP-MS was achieved by the introduction of the collision cell interface in order to dissociate many disturbing argon-based molecular ions, thermalize the ions and neutralize the disturbing argon ions of plasma gas (Ar+). The application of the collision cell in ICP-QMS results in a higher ion transmission, improved sensitivity and better precision of isotope ratio measurements compared to quadrupole ICP-MS without the collision cell [e.g., for 235U/238U approximately 1 (10 microg x L(-1) uranium) 0.07% relative standard deviation (RSD) vs. 0.2% RSD in short-term measurements (n = 5)]. A significant instrumental improvement for ICP-MS is the multicollector device (MC-ICP-MS) in order to obtain a better precision of isotope ratio measurements (with a precision of up to 0.002%, RSD). CE- and HPLC-ICP-MS are used for the separation of isobaric interferences of long-lived radionuclides and stable isotopes by determination of spallation nuclide abundances in an irradiated tantalum target.

  6. A Crease in Gathering 17 of Bodleian MS. Junius 11

    Directory of Open Access Journals (Sweden)

    Alessandra Molinari

    2015-02-01

    Full Text Available This article deals with a crease in gathering 17 of the Old English MS. Junius 11 (Oxford, Bodleian Library, especially with its possible role as evidence that a part of this gathering may have originated as a self-contained booklet before being bound into the MS. After a critical assessment of scholarship on this issue, the article shows that the crease cannot be considered as a piece of evidence for the booklet thesis and that MS. Junius 11 cannot therefore be defined as a composite manuscript.

  7. Comparison of Gluten Extraction Protocols Assessed by LC-MS/MS Analysis.

    Science.gov (United States)

    Fallahbaghery, Azadeh; Zou, Wei; Byrne, Keren; Howitt, Crispin A; Colgrave, Michelle L

    2017-04-05

    The efficiency of gluten extraction is of critical importance to the results derived from any analytical method for gluten detection and quantitation, whether it employs reagent-based technology (antibodies) or analytical instrumentation (mass spectrometry). If the target proteins are not efficiently extracted, the end result will be an under-estimation in the gluten content posing a health risk to people affected by conditions such as celiac disease (CD) and nonceliac gluten sensitivity (NCGS). Five different extraction protocols were investigated using LC-MRM-MS for their ability to efficiently and reproducibly extract gluten. The rapid and simple "IPA/DTT" protocol and related "two-step" protocol were enriched for gluten proteins, 55/86% (trypsin/chymotrypsin) and 41/68% of all protein identifications, respectively, with both methods showing high reproducibility (CV gluten class behaving the same.

  8. MS Windows domēna darbstacijas migrācijas iespējas no MS Windows XP uz MS Windows 7.

    OpenAIRE

    Zariņš, Valdis

    2009-01-01

    Kvalifikācijas darbā tiek aprakstītas MS Windows domēna darbstacijas migrācijas iespējas no MS Windows XP uz MS Windows 7, kā servera operētājsistēmas izmantojot tādus Microsoft produktus, kā Microsoft Windows Server 2003 un Microsoft Windows Server 2008. Kvalifikācijas darba teorētiskaja daļā tiek apskatīti Microsoft Windows 7 priekšrocības un uzlabojumus gan no darbstacijas lietotāja , gan no darbstacijas administratora puses. Ir aprakstītas Microsoft Windows Server 2008 jauninājumu ie...

  9. Rapid profiling and pharmacokinetic studies of major compounds in crude extract from Polygonum multiflorum by UHPLC-Q-TOF-MS and UPLC-MS/MS.

    Science.gov (United States)

    Wang, Linlin; Sang, Mangmang; Liu, Erwei; Banahene, Prince Osei; Zhang, Yi; Wang, Tao; Han, Lifeng; Gao, Xiumei

    2017-06-05

    A reliable, rapid analytical method was established for characterization of constituents in the ethanol extract of Polygonum multiflorum by combining an ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). 131 constituents which including phenolic acids, stilbenes, flavones, anthraquinones, naphthalenes and their derivatives were identified or tentatively identified by using characteristic diagnostic fragment ions and references. The established method was further applied to analyze blood samples, and successfully identified 41 compounds which were absorbed through the gastrointestine in rats after administration the extract of P. multiflorum. Moreover, the pharmacokinetic studies of some major compounds in blood were investigated by using ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method. This study showed a comprehensive research of P. multiflorum, which could provide a meaningful basis for further quality control, pharmacological as well as toxicological researches. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Nonanthocyanin secondary metabolites of black raspberry (Rubus occidentalis L.) fruits: identification by HPLC-DAD, NMR, HPLC-ESI-MS, and ESI-MS/MS analyses.

    Science.gov (United States)

    Paudel, Liladhar; Wyzgoski, Faith J; Scheerens, Joseph C; Chanon, Ann M; Reese, R Neil; Smiljanic, Danijela; Wesdemiotis, Chrys; Blakeslee, Joshua J; Riedl, Kenneth M; Rinaldi, Peter L

    2013-12-11

    Nonanthocyanin secondary metabolites potentially contributing to the antiproliferative bioactivity of black raspberry ( Rubus occidentalis L.) fruits were extracted in ethyl acetate and isolated by semipreparative and analytical HPLC and analyzed by NMR, HPLC-ESI-MS, and ESI-MS/MS techniques. Here we present complete and partial structures of a variety of the chemical entities such as quercetin 3-glucoside, quercetin 3-rutinoside, myricetin glucoside, dihydrokaempferol glucoside, benzoic acid β-d-glucopyranosyl ester, 3,4-dihydroxybenzoic acid, epicatechin, caffeic acid, p-coumaric acid, p-coumaryl glucoside, p-coumaryl sugar ester, ellagic acid, methyl ellagic acid acetylpentose, methyl ellagic acid valerylpentose, trans-piceid, phloretin glucoside (phloridzin), dihydrosinapic acid, salicylic acid β-d-glucopyranosyl ester, a salicylic acid derivative without attached sugar, p-alkylphenyl glucoside, and a citric acid derivative. To our knowledge, 15 of these compounds were not previously reported in black raspberry fruits.

  11. Preclinical development and first-in-human study of ATX-MS-1467 for immunotherapy of MS.

    Science.gov (United States)

    Streeter, Heather B; Rigden, Rachel; Martin, Keith F; Scolding, Neil J; Wraith, David C

    2015-06-01

    The study was designed to test the efficacy of ATX-MS-1467 in a relevant preclinical model and to assess its safety for the treatment of patients with secondary progressive multiple sclerosis (SPMS). ATX-MS-1467 was tested for its ability to suppress experimental autoimmune encephalomyelitis (EAE) in the (Ob x DR2)F1 mouse both before and after disease onset. Safety was assessed by clinical assessment, MRI analysis, and the measurement of immune responses to self- and nonself-antigens in patients with SPMS. ATX-MS-1467 displayed a dose-dependent inhibition of EAE and was more effective than glatiramer acetate in the treatment of ongoing disease in humanized mice. A phase 1 open-label dose-escalating study demonstrated that ATX-MS-1467 was safe and well-tolerated in a group of 6 patients with SPMS, up to a dose of 800 µg. The results of this study support further development of ATX-MS-1467 in a clinical trial powered to investigate the immunologic and clinical benefits of treatment in relapsing-remitting MS. This study provides Class IV evidence that ATX-MS-1467 is safe and tolerated in a group of 6 patients with SPMS.

  12. Comparison of sp-ICP-MS and MDG-ICP-MS for the determination of particle number concentration.

    Science.gov (United States)

    Gschwind, Sabrina; Aja Montes, Maria de Lourdes; Günther, Detlef

    2015-05-01

    In 2011, the European Commission introduced new regulations on how nanomaterials are defined. Since then, researchers have emphasized that more complete characterization of nanoparticles (NPs) includes not just mass and size determinations, but also the determination of the particle number concentrations. In this study, two different sample introduction approaches for the analysis of NP suspensions with inductively coupled plasma mass spectrometry (ICP-MS) were investigated: pneumatic nebulization (sp-ICP-MS) and microdroplet generation (MDG-ICP-MS). These approaches were compared for the determination of particle number concentrations (PNCs) of gold and silver NP suspensions diluted in either ultra-pure water or citrate solution. For accurate sp-ICP-MS analysis, it is crucial to know the transport efficiency of nebulized sample into the plasma. Here, transport efficiencies, measured by the waste collection method, were 11-14 % for Ag suspensions and 9-11 % for Au. In contrast, the droplet transport efficiency of MDG-ICP-MS was 100 %. Analysis by sp-ICP-MS yielded a lower particle number concentration than expected (only 20-40 % of the expected value), whereas MDG-ICP-MS had NP recoveries up to 80 %. This study indicates that NP reference materials are of major importance for particle number determination and detailed results on particle number concentrations for different suspensions with respect to storage time are discussed.

  13. LC-MS/MS determination of pesticide residues in fruits and vegetables.

    Science.gov (United States)

    Stachniuk, Anna; Szmagara, Agnieszka; Czeczko, Renata; Fornal, Emilia

    2017-07-03

    The aim of the research is to evaluate pesticide residue contamination of fresh and frozen fruits and vegetables, agricultural raw material, purchased from Polish farmers for production of frozen fruits and vegetables, and the estimation of the multiresidue method effectiveness expressed as the proportion of pesticides detected in food samples to the total number of pesticides analyzed by multiresidue methods. A total of 144 samples (of black currants, red currants, raspberries, cherries, strawberries, blackberries, cauliflowers and broccoli) were analyzed using LC-MS/MS method for the determination of 60 pesticides. QuEChERS extraction, matrix-matched calibration and dynamic multiple reaction monitoring method were used. Residues of 15 compounds, mainly fungicides and insecticides, were detected in 46 samples. The percentage of samples with residues above the maximum residue levels (MRL) was 15%, whereas samples with residues below MRL were 17%. A total of 13 samples contained more than one pesticide residue. Pesticide residues were detected most often in samples of black currants (50%), broccoli (36.4%), raspberries (29%) and red currants (21.8%). The most frequently detected pesticides were carbendazim and acetamiprid. The proportion of pesticides detected during our study to the total number of analyzed pesticides amounted to 25%. It was compared to literature findings. For three fourth of multiresidue methods, the proportion was below 50% for methods developed for the analysis of less than 100 pesticides, and below 30% for methods developed for the analysis of more than 100 pesticides. It appears that a lot of efforts and means is lost on pesticides never or rarely detected in examined samples. The workload and cost effectiveness of the development and application of multiresidue methods along with the range of pesticides covered by the method should be carefully and thoroughly considered anytime when a new method or workflow is developed. Including non

  14. Screening and Identification of Mitragynine and 7-Hydroxymitragynine in Human Urine by LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Hanzhuo Fu

    2015-05-01

    Full Text Available Kratom is a tree planted in Southeast Asia, including Thailand, Malaysia, Myanmar (Burma and elsewhere in the region. A long history of usage and abuse of kratom has led to the classification of kratom as a controlled substance in its native Thailand and other Southeast Asian countries. However, kratom is not controlled in the United States, and the wide availability of kratom on the Internet and in the streets has led to its emergence as an herbal drug of misuse. With the increasing popularity of kratom, efficient protocols are needed to detect kratom use. In this study, a rapid method for the analysis of kratom compounds, mitragynine and 7-hydroxymitragynine, in human urine has been developed and validated using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS. The chromatographic system employed a 2.6-μm 100 mm × 2.1 mm phenyl-hexyl analytical column and gradient elution with a 0.4-mL/min flow rate of water and acetonitrile as mobile phases. A triple quadrupole mass spectrometer was used as the detector for data acquisition. The analyst was the quantification software. The established method demonstrated linearity of >0.99 for both analytes, and low detection limits were obtained down to 0.002581 ng/mL for mitragynine and 0.06910 ng/mL for 7-hydroxymitragynine. The validated method has been utilized for clinical analysis of urine for the purpose of mitragynine and 7-hydroxymitragynine detection.

  15. Mass spectrometry (LC-MS/MS) identified proteomic biosignatures of breast cancer in proximal fluid

    Science.gov (United States)

    Whelan, Stephen A.; He, Jianbo; Lu, Ming; Souda, Puneet; Saxton, Romaine E.; Faull, Kym F.; Whitelegge, Julian P.; Chang, Helena R.

    2012-01-01

    Summary We have begun an early phase of biomarker discovery in three clinically important types of breast cancer using a panel of human cell lines: HER2 positive, HER2 negative and hormone receptor positive and triple negative (HER2−, ER−, PR−). We identified and characterized the most abundant secreted, sloughed, or leaked proteins released into serum free media from these breast cancer cell lines using a combination of protein fractionation methods before LC-MS/MS mass spectrometry analysis. A total of 249 proteins were detected in the proximal fluid of 7 breast cancer cell lines. The expression of a selected group of high abundance and/or breast cancer specific potential biomarkers including thromobospondin 1, galectin-3 binding protein, cathepsin D, vimentin, zinc-α2-glycoprotein, CD44, and EGFR from the breast cancer cell lines and in their culture media were further validated by Western blot analysis. Interestingly, mass spectrometry identified a cathepsin D protein single-nucleotide polymorphism (SNP) by alanine to valine replacement from the MCF-7 breast cancer cell line. Comparison of each cell line media proteome displayed unique and consistent biosignatures regardless of the individual group classifications demonstrating the potential for stratification of breast cancer. Based on the cell line media proteome, predictive Tree software was able to categorize each cell line as HER2 positive, HER2 negative and hormone receptor positive and triple negative based on only two proteins, muscle fructose 1,6-bisphosphate aldolase and keratin 19. In addition, the predictive Tree software clearly identified MCF-7 cell line overexpresing the HER2 receptor with the SNP cathepsin D biomarker. PMID:22934887

  16. Enantioselective Determination of Polycyclic Musks in River and Wastewater by GC/MS/MS

    Directory of Open Access Journals (Sweden)

    Injung Lee

    2016-03-01

    Full Text Available The separation of chiral compounds is an interesting and challenging topic in analytical chemistry, especially in environmental fields. Enantioselective degradation or bioaccumulation has been observed for several chiral pollutants. Polycyclic musks are chiral and are widely used as fragrances in a variety of personal care products such as soaps, shampoos, cosmetics and perfumes. In this study, the gas chromatographic separation of chiral polycyclic musks, 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclo-penta-γ-2-benzopyrane (HHCB, 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetra-hydronaphthalene (AHTN, 6-acetyl-1,1,2,3,3,5-hexamethylindane (AHDI, 5-acetyl-1,1,2,6-tetramethyl-3-iso-propylindane (ATII, and 6,7-dihydro-1,1,2,3,3-pentamethyl-4(5H-indanone (DPMI was achieved on modified cyclodextrin stationary phase (heptakis (2,3-di-O-methyl-6-O-tert-butyl-dimethylsilyl-β-CD in DV-1701. Separation techniques are coupled to tandem mass spectrometry (MS-MS, as it provides the sensitivity and selectivity needed. River and wastewaters (influents and effluents of wastewater treatment plants (WWTPs in the Nakdong River were investigated with regard to the concentrations and the enantiomeric ratios of polycyclic musks. HHCB was most frequently detected in river and wastewaters, and an enantiomeric enrichment was observed in the effluents of one of the investigated wastewater treatment plants (WWTPs. We reported the contamination of river and wastewaters in Korea by chiral polycyclic musks. The results of this investigation suggest that enantioselective transformation may occur during wastewater treatment.

  17. Profiling of anthocyanins in transgenic purple-fleshed sweet potatoes by HPLC-MS/MS.

    Science.gov (United States)

    Ge, Jingqiu; Hu, Yijie; Wang, Hongxia; Huang, Yuanshe; Zhang, Peng; Liao, Zhihua; Chen, Min

    2017-11-01

    Anthocyanins in purple-fleshed sweet potato (PSP) are beneficial to human health. The leaf color (Lc) gene is a transcription factor involved in regulating anthocyanin biosynthesis. The anthocyanin profiles of wild-type PSP of Ayamurasaki and its three Lc-transgenic lines were investigated by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). In vitro antioxidant activities of wild-type and Lc-transgenic lines, including reducing power activity, DPPH radical scavenging activity, hydroxyl radical scavenging activity, linoleic acid autoxidation inhibition activity, ABTS free radical scavenging activity and oxygen radical absorbance capacity activity, were measured. The results showed that the total anthocyanin contents increased 1.5-1.9 times in three transgenic lines compared with that in wild-type PSP. Seventeen anthocyanins were found in wild-type PSP, while 19 in Lc-transgenic lines including cyanidin-based, peonidin-based and pelargonidin-based anthocyanins. Three pelargonidin-based anthocyanins were detected in three Lc-transgenic lines. Among them, the relative contents of cyanidin-based and pelargonidin-based anthocyanins increased 1.9-2.0 and 3.4-4.5 times respectively, while peonidin-based anthocyanins decreased 1.8-1.9 times in Lc-transgenic lines, compared with wild-type PSP. PSP from wild-type Ayamurasaki and three Lc-transgenic lines exhibited potent antioxidant activities, whereas there was no distinct difference among them. The transgene Lc significantly increased the content of total anthocyanins and remarkably changed the anthocyanin profiles in Ayamurasaki. Such novel and high content of anthocyanins obtained in the Lc-transgenic lines with potent antioxidant activities may provide unique functional products with potential helpful for human health. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  18. Highly sensitive and specific derivatization strategy to profile and quantitate eicosanoids by UPLC-MS/MS

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Ting; Tie, Cai; Wang, Zhe; Zhang, Jin-Lan, E-mail: zhjl@imm.ac.cn

    2017-01-15

    Eicosanoids are signaling molecules mainly oxidized from arachidonic acid (ARA) and eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA). They have attracted increasing attention from the scientists attributing to their essential physiological functions. However, their quantification have long been challenged by the low abundance, high structure similarity, poor stability and limited ionization efficiency. In this paper, an ultra-high performance liquid chromatograph coupled with tandem mass spectrometry (UPLC-MS/MS) strategy was developed for the comprehensive profiling of more than 60 eicosanoids based on an efficient derivatization reagent 2,4-bis(diethylamino)-6-hydrazino-1,3,5-triazine (T3) and general multiple reaction monitoring (MRM) parameters. Carboxylic acid of eicosanoid was converted to amide in 30 min at 4 °C with derivatization yield larger than 99%. Limits of quantitation (LOQs) for derivatized eicosanoids varied from 0.05 to 50 pg depending on their structures. The sensitivities of derivatized eicosanoids were enhanced by 10- to 5000-folds compared to free eicosanoids. Stabilities of T3 modified eicosanoids were also highly improved compared to free eicosanoids. This new method can also be used to quantify eicosanoids in bio-samples using isotopic internal standards with high efficiency and reliability within 19 min. 46 and 50 eicosanoids in rat plasma and heart tissue from control and acute myocardial ischemia (AMI) model rats were respectively profiled and quantitated using this new method. And 24 of 46 and 25 of 50 eicosanoids were found to be significantly changed between control and model groups. The changed eicosanoids related to AMI modeling were further statistically analyzed and interpreted based on eicosanoid metabolism pathway. - Highlights: • Eicosanoids are important signaling molecules. • A highly sensitive and specific derivatization strategy was developed for eicosanoid profiling. • The strategy was employed for

  19. An improved UPLC-MS/MS platform for quantitative analysis of glycerophosphoinositol in mammalian cells.

    Directory of Open Access Journals (Sweden)

    Laura Grauso

    Full Text Available The glycerophosphoinositols constitute a class of biologically active lipid-derived mediators whose intracellular levels are modulated during physiological and pathological cell processes. Comprehensive assessment of the role of these compounds expands beyond the cellular biology of lipids and includes rapid and unambiguous measurement in cells and tissues. Here we describe a sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS method for quantitative analysis of the most abundant among these phosphoinositide derivatives in mammalian cells, the glycerophosphoinositol (GroPIns. The method has been developed in mouse Raw 264.7 macrophages with limits of quantitation at 3 ng/ml. Validation on the same cell line showed excellent response in terms of linear dynamic range (from 3 to 3,000 ng/ml, intra-day and inter-day precision (coefficient of variation ≤ 7.10% and accuracy (between 98.1 and 109.0% in the range 10-320 ng/ml. As proof of concept, a simplified analytical platform based on this method and external calibration was also tested on four stimulated and unstimulated cell lines, including Raw 264.7 macrophages, Jurkat T-cells, A375MM melanoma cells and rat basophilic leukemia RBL-2H3 cells. The results indicate a wide variation in GroPIns levels among different cell lines and stimulation conditions, although the measurements were always in line with the literature. No significant matrix effects were observed thus indicating that the here proposed method can be of general use for similar determinations in cells of different origin.

  20. Quantitative monitoring of corticosteroids in cosmetic products manufactured in Korea using LC-MS/MS.

    Science.gov (United States)

    Nam, Yun Sik; Kwon, Il Keun; Lee, Yeonhee; Lee, Kang-Bong

    2012-07-10

    Some cosmetic products manufactured in Korea for the treatment of eczema, seborrhea and psoriasis have been suspected to contain anti-inflammatory corticosteroids such as prednisolone, hydrocortisone, betamethasone, dexamethasone and triamcinolone acetonide without these ingredients being indicated on the label. Due to their severe side effects such as permenent skin atopy, these corticosteroids have to be monitored in cosmetic products from a forensic point of view. Many cosmetic product samples (N=65) have been collected from both local and online markets in Korea. The corticosteroid content of these samples was analyzed by LC-MS/MS with diagnostic ions (m/z). Linearity was studied with 0.1-10 μg/mL range in all corticosteroids. Good correlation coefficients (r(2)≥0.997) were found and the limits of quantification were 4.68-7.97 ng/mL for each of the corticosteroids. At three different concentrations spanning the linear dynamic ranges, mean recoveries were 97.2-113.5%and precisions (RSD) for intra-day and inter-day analysis were less than 8.9%. Also, accuracy (Bias %) was less than 11.8%. The results showed that between 0.76-0.94 μg/g levels of prednisolone were detected in four cosmetic products and triamcinolone acetonidewas detected with a concentration in the range of 11.5-272 μg/g in nine samples. This fact reveals that some manufacturers have arbitrarily added these corticosteroids in their cosmetic products without indicating them on the label. Thus, these cosmetic products have to be monitored and if proven illegal preparations removed from the market. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  1. Chemical characterization of neonicotinoids in surface waters by high performance liquid chromatography with Tandem Mass Spectrometry (HPLC MS/MS)

    International Nuclear Information System (INIS)

    Amaral, Priscila Oliveira

    2017-01-01

    The present study aimed to develop a method for the determination and validation of a method for the identification and quantification of Neonicotinoids in surface waters collected in the Bauru region, in the state of São Paulo. The analytical techniques studied for the development of this method were the high performance liquid chromatography with tandem mass spectrometry (HPLC - MS / MS), gas chromatography with mass spectrometry (GC / MS) and gas chromatography with electron capture detector (GC / ECD). The class of pesticides Neonicotinoids was chosen for this work because it is related to a sudden disappearance of bees in colonies around the world. This phenomenon is known as Colony Collapse Disorder (CCD) and it is characterized by a rapid loss in the population of adult bees. The Neonicotinoids used in this study were the compounds Clothianidin, Imidacloprid and Thiamethoxam which were banned in their use as pesticides in Europe by Implementing Regulation No. 540/2011. The samples were concentrated using solid phase extraction (SPE) and liquid liquid extraction (LLE) techniques and injected into HPLC-MS / MS, GC / MS and GC / ECD. The GC / ECD and GC / MS techniques were not satisfactory for determination in the water matrix because the detection limit (10 mg L -1 ) is above the maximum allowed by the US Environmental Protection Agency (0.6 μg L -1 ). The HPLC - MS / MS technique using the multiple reaction monitoring (MRM) proved to be adequate for this study because it obtained quantification limits between 5.89 and 8.06 μg L -1 and a linearity between 0.9963 and 0.9999 for the three compounds. (author)

  2. Burnout and Depression in MS1 and MS3 Years: A Comparison of Cohorts at One Medical School.

    Science.gov (United States)

    Elkins, Cinthia; Plante, Kyle P; Germain, Lauren J; Morley, Christopher P

    2017-06-01

    Losses of idealism, empathy, and professionalism over the course of medical education have been described previously in the literature. A 2012 national survey of medical students estimated 37%-44% with burnout, 58% screen positive for depression, and 9.4% with suicidal ideation in the past 12 months. Given changes in related traits, we sought to explore whether burnout and symptoms of depression change over time. The objectives of this study were to assess the differences in medical student burnout and symptoms of depression between first year (MS1) and third-year (MS3) medical students at one medical college. Students were administered the modified Maslach Burnout Inventory (MBI) and the Patient Health Questionnaire-2 (PHQ-2) depression screening instrument as an optional part of a mandatory annual survey administered by the medical college near the end of the academic year (March-May, 2015). Anonymous responses from MS1s and MS3s were analyzed to estimate trends in burnout and possible depression over time and to examine the relationship between the two measures. Possible depression (Total PHQ2 equal or greater than 3) was present in 28.5% of all respondents. MS3s had significantly higher levels of Emotional Exhaustion (EE) and Depersonalization (DP), and lower levels of Personal Accomplishment (PA), than MS1s. When PHQ2, gender, and campus/program effects were controlled via regression, MS3s tended to record EE and DP scores over 2 points higher than MS1s and PA scores over 2 points lower. Regrets about entering medicine were also higher in MS3s in the controlled models. PHQ2 scores were also positively associated with all subscales except for PA. Students appear to be at slightly greater risk for burnout as they temporally progress through medical school. These findings are consistent with similar measurements of decreasing empathy and idealism when comparing cohorts of early versus late medical students.

  3. Quantification of coumarin in cinnamon and woodruff beverages using DIP-APCI-MS and LC-MS.

    Science.gov (United States)

    Krieger, Sonja; Hayen, Heiko; Schmitz, Oliver J

    2013-10-01

    The use of the direct inlet probe-atmospheric-pressure chemical ionization (DIP-APCI) ion source developed in our laboratory coupled to a high resolution Q-TOF MS for the quantitative analysis of coumarin in different cinnamon samples was demonstrated in this study. Extraction of coumarin from various cinnamon samples was followed by DIP-APCI-mass spectrometry (MS) and liquid chromatography (LC)-MS analysis. For quantification, an external calibration with and without the use of stable isotope-labeled coumarin as internal standard was compared. The results obtained by DIP-APCI-MS and LC-MS were in good agreement. Even without the use of an internal standard satisfying linearity (R(2) > 0.997), recovery (94-104% for spiking levels between 100 and 5,000 mg/kg) and intra- and interday repeatability (2.2-13.8%RSD) was demonstrated using DIP-APCI-MS. To reduce the number of samples requiring quantitative analysis, the possibility of semi-quantitative screening of coumarin directly from powdered cinnamon using DIP-APCI-MS was shown. The analysis of woodruff-flavored beverages and cinnamon-flavored chewing gum by DIP-APCI-MS resulted in the formation of an artifact interfering with coumarin detection. As with other ambient ionization methods, special attention has to be paid to possible spectral interferences due to isobaric substances present in the sample matrix or formed from matrix components after ionization. The temperature-programmed vaporization in DIP-APCI-MS combined with the use of stable isotope-labeled coumarin as internal standard helped in recognizing this interference.

  4. Bassey G. Nyengidiki TK Okpani.J

    African Journals Online (AJOL)

    The delivery options for breech presentation are still one of the most disputable issues among Obstetricians worldwide. Objectives: ..... therefore constitutes a major public health problem making regular appraisal of the caesarean delivery very necessary. .... of cephalopelvic disproportion due to early age of marriage, pre-.

  5. CE-MS fingerprinting of Laurencia complex algae (Rhodophyta).

    Science.gov (United States)

    Machín-Sánchez, María; Asensio-Ramos, María; Hernández-Borges, Javier; Gil-Rodríguez, María Candelaria

    2014-03-01

    The use of CE-ESI-MS has been considered as a new chemical strategy for the possible discernment of genera and species of the Laurencia complex. After the selection of the CE-MS and the extraction conditions, a total of 28 specimens of the complex, including different species of four genera (Laurencia, Laurenciella, Palisada, and Osmundea) collected from five intertidal locations on the Island of Tenerife (Canary Islands, Spain) were analyzed. CE-MS fingerprints revealed that CE-MS can be used as a useful tool for these studies in order to assess similarities and differences between them and that it constitutes an important starting point for further studies in the field. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Vitamin D and Multiple Sclerosis (MS): Is There Any Connection?

    Science.gov (United States)

    ... there any connection? Is there any proof that vitamin D supplements can prevent MS or keep symptoms of ... School have suggested that maintaining adequate levels of vitamin D may have a protective effect and lower the ...

  7. When a Parent Has MS: A Teenager's Guide

    Science.gov (United States)

    ... I transformed my father having MS into personal independence and never letting an obstacle, no matter its ... that they will have to shelve plans for college. They assume it will cost too much on ...

  8. Practical guide to ICP-MS: a tutorial for beginners

    National Research Council Canada - National Science Library

    Thomas, Robert

    2013-01-01

    ... for those with limited knowledge of the technique. Written by an insider with more than 20 years experience in product development, customer support, and technical marketing for an ICP-MS instrument vendor, the book highlights this powerful ultra...

  9. Comparison of clonazepam compliance by measurement of urinary concentration by immunoassay and LC-MS/MS in pain management population.

    Science.gov (United States)

    West, Robert; Pesce, Amadeo; West, Cameron; Crews, Bridgit; Mikel, Charles; Almazan, Perla; Rosenthal, Murray; Latyshev, Sergey

    2010-01-01

    Physicians determine patient compliance with their medications by use of urine drug testing. It is known that measurement of benzodiazepines is limited by immunoassay specificity and cutoff limits and therefore does not offer physicians an accurate picture of their patients' compliance with these medications. A few studies have used lower cutoffs to demonstrate patient compliance. To define more appropriate cutoffs for compliance monitoring of patients prescribed clonazepam as determined using immunoassay and liquid chromatography-tandem mass spectrometry (LC-MS/MS). A diagnostic accuracy study of the urinary excretion of clonazepam. Millennium Laboratories performed measurements on the urinary excretion of pain patients prescribed clonazepam as the indicator test. This benzodiazepine was chosen because it forms one major metabolite, 7-aminoclonazepam which is specific for that drug. Patients whose only benzodiazepine medication was clonazepam were selected as the test population. The Millennium Laboratories test database was filtered first to select patients on clonazepam, then a second filter was used to eliminate patients with any other listed benzodiazepine medications. Samples were tested using the Microgenics DRI benzodiazepine assay with a 200 ng/mL cutoff. The same samples were quantitatively assessed for 7-aminoclonazepam by LC-MS/MS with a cutoff of 40 ng/mL. The results from the immunoassay were scored as positive or negative while the quantitative results from the LC-MS/MS were also scored as positive or negative depending upon their concentration. Samples from 180 patients met these medication criteria. The positivity rates were 21% (38 samples) by immunoassay. The positivity rate was 70% (126 samples) if the LC-MS/MS cutoff was set at 200 ng/mL. However, the positivity rate was 87% (157 samples) if the LC-MS/MS was set at 40 ng/mL. Concentration distributions revealed a significant fraction (7%) in the 40 - 100 ng/mL range. A limitation of the study

  10. METHOD 544. DETERMINATION OF MICROCYSTINS AND NODULARIN IN DRINKING WATER BY SOLID PHASE EXTRACTION AND LIQUID CHROMATOGRAPHY/TANDEM MASS SPECTROMETRY (LC/MS/MS)

    Science.gov (United States)

    Method 544 is an accurate and precise analytical method to determine six microcystins (including MC-LR) and nodularin in drinking water using solid phase extraction and liquid chromatography tandem mass spectrometry (SPE-LC/MS/MS). The advantage of this SPE-LC/MS/MS is its sensi...

  11. Social cognition in pediatric-onset multiple sclerosis (MS).

    Science.gov (United States)

    Charvet, L E; Cleary, R E; Vazquez, K; Belman, A L; Krupp, L B

    2014-10-01

    Pediatric-onset multiple sclerosis (MS) patients represent a subpopulation who are diagnosed during the course of development. Social cognitive deficits have recently been recognized in adults with MS. It is critical to identify whether these youngest patients with the disorder are also at risk. To determine whether pediatric-onset MS is associated with social cognitive deficits. Consecutively-recruited participants with pediatric-onset MS were compared to a group of age- and gender-matched healthy controls on Theory of Mind (ToM) task performance. Tasks measured facial affect recognition (Reading the Mind in the Eyes Test), detecting social faux pas (Faux Pas Test), and understanding the perspective of another (False Beliefs Task). Twenty-eight (28) pediatric-onset MS participants (median age 17 years) and 32 healthy controls (median age 16 years) completed the study. The MS participants performed worse than controls on all three ToM tasks: Reading the Mind in the Eyes Test (p = 0.008), the Faux Pas Test (p = 0.009), and the False Beliefs Task (p = 0.06). While more MS than control participants were impaired on a measure of information processing speed (the Symbol Digit Modalities Test; 38% versus 6%), it did not account for the differences in ToM performance. Social cognition may represent an area of cognitive functioning affected by MS in the pediatric-onset population. These processes are especially important to study in younger patients as they may have long range implications for social adjustment, employment, and well-being. © The Author(s) 2014.

  12. GC-MS investigation of tropane alkaloids in Datura stramonium.

    Science.gov (United States)

    Philipov, Stefan; Berkov, Strahil

    2002-01-01

    Alkaloids, GS-MS, Datura stramonium The alkaloid spectrum in roots, leaves and seeds of Datura stramonium L. was investigated by GC-MS. Twenty-nine tropane alkaloids are detected. Twelve of them are new constituents for the species and the two tropane esters 3-(3'-acetoxytropoyloxy)tropane (21) and 3-(2'-hydroxytropoyloxy)tropane (26) are described for the first time.

  13. Mental Health Comorbidity in MS: Depression, Anxiety, and Bipolar Disorder.

    Science.gov (United States)

    Turner, Aaron P; Alschuler, Kevin N; Hughes, Abbey J; Beier, Meghan; Haselkorn, Jodie K; Sloan, Alicia P; Ehde, Dawn M

    2016-12-01

    Among individuals with multiple sclerosis (MS), mental health comorbidities play a significant role in contributing to secondary disability and detracting from quality of life. This review examines current evidence surrounding three mental health issues of particular relevance to MS: depression, anxiety, and bipolar disorder. We review what is known of the prevalence, correlates, screening mechanisms, and current treatment of each issue and provide recommendations for future areas of research.

  14. Cognitive Impairment in MS Linked to Structural and Functional Connectivity

    Science.gov (United States)

    2016-10-01

    neuro-axonal loss and demyelination that occurs in MS 1. A recent meta-analysis of 18 studies of regional grey matter loss in RRMS found that, in...regions were lower in MS subjects than controls. From dMRI, estimates of the directionality, water diffusion can be calculated throughout the brain...resulting in a higher FA since water molecules may travel in many different directions. In disease, if one or more of those pathways are destroyed

  15. Butter and butter oil classification by PTR-MS

    OpenAIRE

    Ruth, van, S.M.; Koot, A.H.; Akkermans, W.; Araghipour, N.; Rozijn, M.; Baltussen, M.A.H.; Wisthaler, A.; Mark, T.D.; Frankhuizen, R.

    2008-01-01

    The potential of proton transfer reaction mass spectrometry (PTR-MS) as a tool for classification of milk fats was evaluated in relation to quality and authentication issues. Butters and butter oils were subjected to heat and off-flavouring treatments in order to create sensorially defective samples. The effect of the treatments was evaluated by means of PTR-MS analysis, sensory analysis and classical chemical analysis. Subsequently, partial least square-discriminant analysis models (PLS-DA) ...

  16. Demographic characteristics of MS patients in Poland's upper Silesia region.

    Science.gov (United States)

    Pierzchala, Krystyna; Adamczyk-Sowa, Monika; Dobrakowski, Pawel; Kubicka-Baczyk, Katarzyna; Niedziela, Natalia; Sowa, Pawel

    2015-05-01

    In Poland, no national registry of MS patients has yet been introduced. So far, no demographic studies have been conducted in patients with MS in Upper Silesia. The aim of the present study was to evaluate, for the first time, a selected demographic and clinical parameters in MS patients from the Upper Silesia region and compare these characteristics with previously published data from other regions of Poland. 640 patients with clinically defined MS, were prospectively and randomly selected for the study. Social, socio-economic, and demographic data were obtained through a questionnaire study. All subjects performed a self-assessment of their health condition using EQ-5D and EQ-VAS version questionnaires. The ratio of women to men was 2.18. The average age of onset was 29.6 ± 11.1 years; the disease duration was 7.9 ± 4.5 years. The relapsing-remitting form of MS was diagnosed in 73.12%. In 71.25% the onset was monofocal and in 28.75% multifocal disease onset was observed. Among the studied population 339 (52.97%) patients were still employed. A mean EQ-VAS score of 66.11 ± 20.12 was calculated. Results from our study identify for the first time the demographic and clinical characteristics of the Upper Silesia MS population.

  17. Post-acquisition data mining techniques for LC-MS/MS-acquired data in drug metabolite identification.

    Science.gov (United States)

    Dhurjad, Pooja Sukhdev; Marothu, Vamsi Krishna; Rathod, Rajeshwari

    2017-08-01

    Metabolite identification is a crucial part of the drug discovery process. LC-MS/MS-based metabolite identification has gained widespread use, but the data acquired by the LC-MS/MS instrument is complex, and thus the interpretation of data becomes troublesome. Fortunately, advancements in data mining techniques have simplified the process of data interpretation with improved mass accuracy and provide a potentially selective, sensitive, accurate and comprehensive way for metabolite identification. In this review, we have discussed the targeted (extracted ion chromatogram, mass defect filter, product ion filter, neutral loss filter and isotope pattern filter) and untargeted (control sample comparison, background subtraction and metabolomic approaches) post-acquisition data mining techniques, which facilitate the drug metabolite identification. We have also discussed the importance of integrated data mining strategy.

  18. A rapid LC-MS/MS quantification of peramivir using a simple and inexpensive sample precipitation: application to PK.

    Science.gov (United States)

    Wang, Zhan-Zhang; Zhang, Ming; Shang, De-Wei; Ni, Xiao-Jia; Hu, Jin-Qing; Qiu, Chang; Wen, Yu-Guan

    2015-01-01

    Peramivir is a newly approved selective neuraminidase inhibitor designed to inhibit influenza virus infection. We report a robust and sensitive method utilizing simple precipitation extraction with LC-MS/MS for the high-throughput quantification. Addition of 0.06 M of ammonium formate and 0.1% formic acid in mobile phase could help reduce the matrix effect. This method uses 100 µl of plasma and covers a linear concentration range from 5 to 10,000 ng/ml. Other validation parameters are also evaluated and meet regulatory expectations by US FDA guidelines. The developed HPLC-MS/MS method has been successfully applied to support a clinical pharmacokinetic study. The strategy presented here can be applied elsewhere and may be useful for other amphiphilic drugs.

  19. Quantitation of Buprenorphine, Norbuprenorphine, Buprenorphine Glucuronide, Norbuprenorphine Glucuronide, and Naloxone in Urine by LC-MS/MS.

    Science.gov (United States)

    Marin, Stephanie J; McMillin, Gwendolyn A

    2016-01-01

    Buprenorphine is an opioid drug that has been used to treat opioid dependence on an outpatient basis, and is also prescribed for managing moderate to severe pain. Some formulations of buprenorphine also contain naloxone to discourage misuse. The major metabolite of buprenorphine is norbuprenorphine. Both compounds are pharmacologically active and both are extensively metabolized to their glucuronide conjugates, which are also active metabolites. Direct quantitation of the glucuronide conjugates in conjunction with free buprenorphine, norbuprenorphine, and naloxone in urine can distinguish compliance with prescribed therapy from specimen adulteration intended to mimic compliance with prescribed buprenorphine. This chapter quantitates buprenorphine, norbuprenorphine, their glucuronide conjugates and naloxone directly in urine by liquid chromatography tandem mass spectrometry (LC-MS/MS). Urine is pretreated with formic acid and undergoes solid phase extraction (SPE) prior to analysis by LC-MS/MS.

  20. Screening for Anabolic Steroids in Urine of Forensic Cases Using Fully Automated Solid Phase Extraction and LC–MS-MS

    DEFF Research Database (Denmark)

    Andersen, David Wederkinck; Linnet, Kristian

    2014-01-01

    A screening method for 18 frequently measured exogenous anabolic steroids and the testosterone/epitestosterone (T/E) ratio in forensic cases has been developed and validated. The method involves a fully automated sample preparation including enzyme treatment, addition of internal standards...... and solid phase extraction followed by analysis by liquid chromatography-tandem mass spectrometry (LC-MS-MS) using electrospray ionization with adduct formation for two compounds. Urine samples from 580 forensic cases were analyzed to determine the T/E ratio and occurrence of exogenous anabolic steroids...... were seen in the majority of cases. The method presented serves as a fast and automated screening procedure, proving the suitability of LC-MS-MS for analyzing anabolic steroids....