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Sample records for rats simultaneous analysis

  1. Simultaneous analysis of diazepam and its metabolites in rat plasma and brain tissue by HPLC-UV and SPE.

    Science.gov (United States)

    Mercolini, Laura; Mandrioli, Roberto; Iannello, Carmelina; Matrisciano, Francesco; Nicoletti, Ferdinando; Raggi, Maria Augusta

    2009-11-15

    Diazepam is frequently used as an adjuvant during antidepressant therapy. Recently, some studies have suggested that the treatment with benzodiazepines could have different efficacy in depressed patients as opposed to non-depressed ones. To clarify the matter, a study is currently underway, regarding the drug metabolism in rats. In order to obtain a more complete and significant set of data, the main diazepam metabolites have also been considered, namely: nordiazepam, temazepam and oxazepam. A feasible and reliable HPLC method has been developed for the simultaneous determination of these compounds in plasma and brain tissue of rats. The method has been applied to "normal" rats and to genetic rat models of depression in order to estimate drug metabolism in different breeds. Analyte separation was achieved on a C8 reversed phase column using an acidic phosphate buffer/acetonitrile mixture as the mobile phase. The detection wavelength was 238 nm. An original sample pre-treatment, based on solid-phase extraction (SPE) was developed in order to eliminate endogenous interference, using only 250 microL of matrix (brain homogenate or plasma) for a complete analysis. The method has been validated with good results in terms of precision, extraction yield, sensitivity, selectivity and accuracy on both matrices and has been successfully applied to samples from some rats subjected to the preliminary study. The obtained data will hopefully contribute to the clarification of possible differences between depressed and non-depressed subjects with respect to benzodiazepine biotransformation.

  2. Simultaneous UPLC-MS/MS analysis of native catechins and procyanidins and their microbial metabolites in intestinal contents and tissues of male Wistar Furth inbred rats.

    Science.gov (United States)

    Goodrich, Katheryn M; Neilson, Andrew P

    2014-05-01

    Procyanidins have been extensively investigated for their potential health protective activities. However, the potential bioactivities of procyanidins are limited by their poor bioavailability. The majority of the ingested dose remains unabsorbed and reaches the colon where extensive microbial metabolism occurs. Most existing analytical methods measure either native compounds (catechins and procyanidins), or their microbial metabolites. The objectives of this study were to develop a high-throughput extraction and UPLC-MS/MS method for simultaneous measurement of both native procyanidins and their metabolites, facilitating high-throughput analysis of native and metabolite profiles in various regions of the colon. The present UPLC-MS/MS method facilitates simultaneous resolution and detection of authentic standards of 14 native catechin monomers and procyanidins, as well as 24 microbial metabolites. Detection and resolution of an additional 3 procyanidin dimers and 10 metabolites for which standards were not available was achieved. Elution and adequate resolution of both native compounds and metabolites were achieved within 10min. The intraday repeatability for native compounds was between 1.1 and 16.5%, and the interday repeatability for native compounds was between 2.2 and 25%. Intraday and interday repeatability for metabolites was between 0.6 and 24.1% and 1 and 23.9%, respectively. Observed lower limits of quantification for native compounds were ∼9-350fmol on-column, and for the microbial metabolites were ∼0.8-12,000fmol on-column. Observed lower limits of detection for native compounds were ∼4.5-190fmol on-column, and for metabolites were 0.304-6020fmol on-column. For native monomers and procyanidins, extraction recoveries ranged from 38 to 102%. Extraction recoveries for the 9 microbial metabolites tested ranged from 41 to 95%. Data from tissue analysis of rats gavaged with grape seed extract indicate fairly high accumulation of native compounds

  3. Development and validation of bioanalytical UHPLC-UV method for simultaneous analysis of unchanged fenofibrate and its metabolite fenofibric acid in rat plasma: Application to pharmacokinetics

    Directory of Open Access Journals (Sweden)

    Rayan G. Alamri

    2017-01-01

    Full Text Available A simple, precise, selective and fast ultra-high performance liquid chromatography (UHPLC-UV method has been developed and validated for the simultaneous determination of a lipid regulating agent fenofibrate and its metabolite fenofibric acid in rat plasma. The chromatographic separation was carried out on a reversed-phase Acquity® BEH C18 column using methanol–water (65:35, v/v as the mobile phase. The isocratic flow was 0.3 ml/min with rapid run time of 2.5 min and UV detection was at 284 nm. The method was validated over a concentration range of 100–10000 ng/ml (r2 ⩾ 0.9993. The selectivity, specificity, recovery, accuracy and precision were validated for determination of fenofibrate/fenofibric acid in rat plasma. The lower limits of detection and quantitation of the method were 30 and 90 ng/ml for fenofibrate and 40 and 100 ng/ml for fenofibric acid, respectively. The within and between-day coefficients of variation were less than 5%. The validated method has been successfully applied to measure the plasma concentrations in pharmacokinetics study of fenofibrate in an animal model to illustrate the scope and application of the method.

  4. A reversed-phase HPLC-UV assay for simultaneous analysis of EGCG and ECG of tea polyphenols in rat plasma

    Institute of Scientific and Technical Information of China (English)

    FU Ting; LIANG Jun; HAN Guo-zhu; L(U) Li; LI Nan

    2008-01-01

    Objective To develop a simple and specific reversed-phase HPLC-UV method for simultaneous determination of (-) Epigallocatechin-3-gallate (EGCG) and (-) Epicatechin-3-gallate (ECG), the main active ingredients of tea polyphenols (TP), in rat plasma. Methods EGCG and ECG were eluted on a Kromasil C18 analytical column (150 mm×4.6 mm, 5μm) protected by a C18 pre-column (4.6 mm×20 mm, 10μm) with a linear gradient mobile phase composed of CH3CN (A)-0.1% citric acid (B), which was run from initial 14 % A and 86 % B to 20 % A and 80 % B at a flow rate of 1.0 mL·min-1 in 14 min, then changed to 14% A and 86% B at a gradient flow rate of 1.0-1.5 mL·min-1 during 14-18 min, and then maintained until 22 min at a gradient flow rate of 1.5-1.0 mL·min-1. The UV detector was set at 280 nm. Plasma samples (200 μL each) were prepared by liquid-liquid extraction procedure with double volumes of EtoAc and then evaporation of organic phase under N2 stream to dry, followed by reconstitution with 100 μL of 20 % CH3CN aqueous solution. The peak area ratios of analytes to vanillin as internal standard vs concentration of analytes to construct calibration curves. Results The HPLC resulted in base-line separation of vanillin, EGCG, ECG and other components; there was no interference from blank plasma. The linear range was 0.5-300 μg·mL-1 for EC, CG (r=0.9999) and 0.1-60 μg·mL-1 for ECG (r = 0.9999). The intra-and inter-day precision (RSD) was better than 6.1% and 12.6%, respectively, and the average accuracy was between 86.25%-103.14%. The extraction recovery of EGCG and ECG was 79.80%-84.64% and 75.22 %-91.39 %, respectively. The plasma samples were stable for at least 30 days at -20 ℃ and 8 h at room temperature;EGCG, ECG and IS stock solutions 2 months at -20 ℃, and the EtoAc-extracted plasma samples 24 h at 4 ℃. Application of the method to the determination of EGCG and ECG in plasma of rats receiving iv 100 mg·kg-1 of TP showed that these 2 compounds

  5. Early Effect of Amyloid β-Peptide on Hippocampal and Serum Metabolism in Rats Studied by an Integrated Method of NMR-Based Metabolomics and ANOVA-Simultaneous Component Analysis

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    Du, Yao; Xia, Huanhuan; Zhao, Liangcai; Hu, Wenyi; Bai, Guanghui

    2017-01-01

    Amyloid β (Aβ) deposition has been implicated in the pathogenesis of Alzheimer's disease. However, the early effect of Aβ deposition on metabolism remains unclear. In the present study, thus, we explored the metabolic changes in the hippocampus and serum during first 2 weeks of Aβ25–35 injection in rats by using an integrated method of NMR-based metabolomics and ANOVA-simultaneous component analysis (ASCA). Our results show that Aβ25–35 injection, time, and their interaction had statistically significant effects on the hippocampus and serum metabolome. Furthermore, we identified key metabolites that mainly contributed to these effects. After Aβ25–35 injection from 1 to 2 weeks, the levels of lactate, N-acetylaspartate, creatine, and taurine were decreased in rat hippocampus, while an increase in lactate and decreases in LDL/VLDL and glucose were observed in rat serum. Therefore, we suggest that the reduction in energy and lipid metabolism as well as an increase in anaerobic glycolysis may occur at the early stage of Aβ25–35 deposition.

  6. Early Effect of Amyloid β-Peptide on Hippocampal and Serum Metabolism in Rats Studied by an Integrated Method of NMR-Based Metabolomics and ANOVA-Simultaneous Component Analysis.

    Science.gov (United States)

    Du, Yao; Zheng, Hong; Xia, Huanhuan; Zhao, Liangcai; Hu, Wenyi; Bai, Guanghui; Yan, Zhihan; Gao, Hongchang

    2017-01-01

    Amyloid β (Aβ) deposition has been implicated in the pathogenesis of Alzheimer's disease. However, the early effect of Aβ deposition on metabolism remains unclear. In the present study, thus, we explored the metabolic changes in the hippocampus and serum during first 2 weeks of Aβ25-35 injection in rats by using an integrated method of NMR-based metabolomics and ANOVA-simultaneous component analysis (ASCA). Our results show that Aβ25-35 injection, time, and their interaction had statistically significant effects on the hippocampus and serum metabolome. Furthermore, we identified key metabolites that mainly contributed to these effects. After Aβ25-35 injection from 1 to 2 weeks, the levels of lactate, N-acetylaspartate, creatine, and taurine were decreased in rat hippocampus, while an increase in lactate and decreases in LDL/VLDL and glucose were observed in rat serum. Therefore, we suggest that the reduction in energy and lipid metabolism as well as an increase in anaerobic glycolysis may occur at the early stage of Aβ25-35 deposition.

  7. Beamformer for simultaneous magnetoencephalography and electroencephalography analysis

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    Ko, Seokha; Jun, Sung Chan

    2010-05-01

    We proposed the beamformer for simultaneous magnetoencephalography (MEG)/electroencephalography (EEG) analysis which has the synergy effects such as high spatial resolution, low localization bias and robustness for orientation of brain sources. Through Monte Carlo simulation study, it was found that the localization performance of our proposed beamformer was far superior to those of MEG-only and EEG-only. For the given specific sensor geometry (160 MEG, 50 EEG sensors), we investigated comparative localization performance of our proposed beamformer over various weighting factors of MEG data, while weighting factor of EEG keeps fixed. Furthermore, we demonstrated its capability for simulated two dipole problem and empirical somatosensory median nerve stimulation data.

  8. Simultaneous MRI and PET imaging of a rat brain

    Science.gov (United States)

    Raylman, Raymond R.; Majewski, Stan; Lemieux, Susan K.; Sendhil Velan, S.; Kross, Brian; Popov, Vladimir; Smith, Mark F.; Weisenberger, Andrew G.; Zorn, Carl; Marano, Gary D.

    2006-12-01

    Multi-modality imaging is rapidly becoming a valuable tool in the diagnosis of disease and in the development of new drugs. Functional images produced with PET fused with anatomical structure images created by MRI will allow the correlation of form with function. Our group is developing a system to acquire MRI and PET images contemporaneously. The prototype device consists of two opposed detector heads, operating in coincidence mode. Each MRI-PET detector module consists of an array of LSO detector elements coupled through a long fibre optic light guide to a single Hamamatsu flat panel position-sensitive photomultiplier tube (PSPMT). The use of light guides allows the PSPMTs to be positioned outside the bore of a 3T MRI scanner where the magnetic field is relatively small. To test the device, simultaneous MRI and PET images of the brain of a male Sprague Dawley rat injected with FDG were successfully obtained. The images revealed no noticeable artefacts in either image set. Future work includes the construction of a full ring PET scanner, improved light guides and construction of a specialized MRI coil to permit higher quality MRI imaging.

  9. Simultaneous MRI and PET imaging of a rat brain

    Energy Technology Data Exchange (ETDEWEB)

    Raylman, Raymond R [Center for Advanced Imaging, Department of Radiology, Box 9236, West Virginia University, Morgantown, WV (United States); Majewski, Stan [Thomas Jefferson National Accelerator Facility, 12000 Jefferson Ave., Newport News, VA (United States); Lemieux, Susan K [Center for Advanced Imaging, Department of Radiology, Box 9236, West Virginia University, Morgantown, WV (United States); Velan, S Sendhil [Center for Advanced Imaging, Department of Radiology, Box 9236, West Virginia University, Morgantown, WV (United States); Kross, Brian [Thomas Jefferson National Accelerator Facility, 12000 Jefferson Ave., Newport News, VA (United States); Popov, Vladimir [Thomas Jefferson National Accelerator Facility, 12000 Jefferson Ave., Newport News, VA (United States); Smith, Mark F [Thomas Jefferson National Accelerator Facility, 12000 Jefferson Ave., Newport News, VA (United States); Weisenberger, Andrew G [Thomas Jefferson National Accelerator Facility, 12000 Jefferson Ave., Newport News, VA (United States); Zorn, Carl [Thomas Jefferson National Accelerator Facility, 12000 Jefferson Ave., Newport News, VA (United States); Marano, Gary D [Center for Advanced Imaging, Department of Radiology, Box 9236, West Virginia University, Morgantown, WV (United States)

    2006-12-21

    Multi-modality imaging is rapidly becoming a valuable tool in the diagnosis of disease and in the development of new drugs. Functional images produced with PET fused with anatomical structure images created by MRI will allow the correlation of form with function. Our group is developing a system to acquire MRI and PET images contemporaneously. The prototype device consists of two opposed detector heads, operating in coincidence mode. Each MRI-PET detector module consists of an array of LSO detector elements coupled through a long fibre optic light guide to a single Hamamatsu flat panel position-sensitive photomultiplier tube (PSPMT). The use of light guides allows the PSPMTs to be positioned outside the bore of a 3T MRI scanner where the magnetic field is relatively small. To test the device, simultaneous MRI and PET images of the brain of a male Sprague Dawley rat injected with FDG were successfully obtained. The images revealed no noticeable artefacts in either image set. Future work includes the construction of a full ring PET scanner, improved light guides and construction of a specialized MRI coil to permit higher quality MRI imaging.

  10. Simultaneity Analysis In A Wireless Sensor Network

    Directory of Open Access Journals (Sweden)

    Malović Miodrag

    2015-06-01

    Full Text Available An original wireless sensor network for vibration measurements was designed. Its primary purpose is modal analysis of vibrations of large structures. A number of experiments have been performed to evaluate the system, with special emphasis on the influence of different effects on simultaneity of data acquired from remote nodes, which is essential for modal analysis. One of the issues is that quartz crystal oscillators, which provide time reading on the devices, are optimized for use in the room temperature and exhibit significant frequency variations if operated outside the 20–30°C range. Although much research was performed to optimize algorithms of synchronization in wireless networks, the subject of temperature fluctuations was not investigated and discussed in proportion to its significance. This paper describes methods used to evaluate data simultaneity and some algorithms suitable for its improvement in small to intermediate size ad-hoc wireless sensor networks exposed to varying temperatures often present in on-site civil engineering measurements.

  11. Scaling in ANOVA-simultaneous component analysis.

    Science.gov (United States)

    Timmerman, Marieke E; Hoefsloot, Huub C J; Smilde, Age K; Ceulemans, Eva

    In omics research often high-dimensional data is collected according to an experimental design. Typically, the manipulations involved yield differential effects on subsets of variables. An effective approach to identify those effects is ANOVA-simultaneous component analysis (ASCA), which combines analysis of variance with principal component analysis. So far, pre-treatment in ASCA received hardly any attention, whereas its effects can be huge. In this paper, we describe various strategies for scaling, and identify a rational approach. We present the approaches in matrix algebra terms and illustrate them with an insightful simulated example. We show that scaling directly influences which data aspects are stressed in the analysis, and hence become apparent in the solution. Therefore, the cornerstone for proper scaling is to use a scaling factor that is free from the effect of interest. This implies that proper scaling depends on the effect(s) of interest, and that different types of scaling may be proper for the different effect matrices. We illustrate that different scaling approaches can greatly affect the ASCA interpretation with a real-life example from nutritional research. The principle that scaling factors should be free from the effect of interest generalizes to other statistical methods that involve scaling, as classification methods.

  12. A brief analysis on the Simultaneous Interpretation

    Institute of Scientific and Technical Information of China (English)

    Qi Wang

    2016-01-01

    Simultaneous interpretation (SI) is an intellectual activity which requires interpreter to translate form one language into another simultaneously. As the most challenged field of interpretation, SI is considered to be the highest level of language interpreting for it requires special quality of skills which can never be acquired without systematic and sophisticated training.

  13. Simultaneous analysis of D-alanine, D-aspartic acid, and D-serine using chiral high-performance liquid chromatography-tandem mass spectrometry and its application to the rat plasma and tissues.

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    Karakawa, Sachise; Shimbo, Kazutaka; Yamada, Naoyuki; Mizukoshi, Toshimi; Miyano, Hiroshi; Mita, Masashi; Lindner, Wolfgang; Hamase, Kenji

    2015-11-10

    A highly sensitive and selective chiral LC-MS/MS method for D-alanine, D-aspartic acid and D-serine has been developed using the precolumn derivatization reagents, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AccQ-Tag) or p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS). The thus N-tagged enantiomers of the derivatized amino acids were nicely separated within 20min using the cinchona alkaloid-based zwittterionic ion-exchange type enantioselective column, Chiralpak ZWIX(+). The selected reaction monitoring was applied for detecting the target d-amino acids in biological matrices. By using the present chiral LC-MS/MS method, the three d-amino acids and their l-forms could be simultaneously determined in the range of 0.1-500nmol/mL. Finally, the technique was successfully applied to rat plasma and tissue samples.

  14. Analysis of Errors Encountered in Simultaneous Interpreting

    Institute of Scientific and Technical Information of China (English)

    方峥

    2015-01-01

    I.Introduction1.1 Definition of an error An error happens when the interpreter’s delivery affects the communicative impact of the speaker’s message,including semantic inaccuracies and inaccuracies of presentation.Along with the development of simultaneous interpreting,there has been a number of professional interpreters and linguists present their definitions and points of views about the errors

  15. Responding by exclusion in Wistar rats in a simultaneous visual discrimination task.

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    Felipe de Souza, Matheus; Schmidt, Andréia

    2014-11-01

    Responding by exclusion is to select a correct alternative by rejecting other potential alternatives. Studies describe this ability in some mammals and birds. However, this type of performance has not been reported in rodents. The aim of this study was to verify the occurrence of responding by exclusion in Wistar rats after a baseline of simple simultaneous visual discrimination. Six male Wistar learned nose-poking tunnels displaying visual stimuli (projected geometric shapes) in an operant chamber. After establishing the simultaneous discrimination baseline, three probe sessions were conducted. In each session, there was a novelty-control probe (a new stimulus was presented together with a stimulus trained as the S(+)) and an exclusion probe (a second new stimulus was presented simultaneously with a stimulus trained as the S(-)). Only one rat responded to the new stimulus in one of the three novelty probe trials. Four rats responded to the three new stimuli and one responded to the new stimulus in two of the three exclusion probes. One subject responded to the S(-) in all the exclusion probes. Five of the six subjects were therefore able to choose the new stimulus, rejecting stimuli trained as the S(-). This is the first experimental evidence for performance by exclusion in rats.

  16. Methodology developed for the simultaneous measurement of bone formation and bone resorption in rats based on the pharmacokinetics of fluoride.

    Science.gov (United States)

    Lupo, Maela; Brance, Maria Lorena; Fina, Brenda Lorena; Brun, Lucas Ricardo; Rigalli, Alfredo

    2015-01-01

    This paper describes a novel methodology for the simultaneous estimation of bone formation (BF) and resorption (BR) in rats using fluoride as a nonradioactive bone-seeker ion. The pharmacokinetics of flouride have been extensively studied in rats; its constants have all been characterized. This knowledge was the cornerstone for the underlying mathematical model that we used to measure bone fluoride uptake and elimination rate after a dose of fluoride. Bone resorption and formation were estimated by bone fluoride uptake and elimination rate, respectively. ROC analysis showed that sensitivity, specificity and area under the ROC curve were not different from deoxypiridinoline and bone alkaline phosphatase, well-known bone markers. Sprague-Dawley rats with modified bone remodelling (ovariectomy, hyper, and hypocalcic diet, antiresorptive treatment) were used to validate the values obtained with this methodology. The results of BF and BR obtained with this technique were as expected for each biological model. Although the method should be performed under general anesthesia, it has several advantages: simultaneous measurement of BR and BF, low cost, and the use of compounds with no expiration date.

  17. [Deuterated surrogate and UPLC-MS/MS method for simultaneous determination of 22 endocannabinoids in ovariectomized rat plasma].

    Science.gov (United States)

    Xiang, Shi-Xie; Xu, Ying; Zhu, Jing-Jing; Wang, Zhi-Min; Zhang, Dong; Chen, Liang-Mian; Feng, Wei-Hong

    2016-11-01

    A new method based on ultraperformance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for the simultaneous determination of 22 endocannabinoids(eCBs) and relevant compounds in ovariectomized rat plasma. After being extracted by solid-phase column(SPE), the plasma samples were detected by using UPLC-MS/MS. Analysis was carried out with ACQUITY UPLC BEH C₁₈ column. The mobile phase was 0.1% acetic acid solution(A)-acetonitrile and isopropanol(9∶1, B) for the gradient elution. In the positive ion multiple reaction monitoring(MRM) mode, deuterated reagents were taken as standard alternatives to calculate recoveries and simultaneously quantify 22 endocannabinoids. The established method provided a good linearity for the 22 eCBs, and their linearly dependent coefficients were all higher than 0.99. The limits of quantitation(LOQs) ranged from 0.089 6 to 1.965 2 nmol•L-1. Relative recoveries of 5 deuterated surrogates ranged between 11.40% and 129.9%. The repeatability study results showed that RSD was all less than 8.0%. The established method could be used to analyze PGF2a EA, AEA and other endogenous cannabinoids in plasma samples of ovariectomized rats. In summary, this method was proved to boast a high sensitivity, repeatability and practicability, and thus could be used in rat plasma lipid metabolomics study. Copyright© by the Chinese Pharmaceutical Association.

  18. Simultaneous bilateral cataract surgery: economic analysis; Helsinki Simultaneous Bilateral Cataract Surgery Study Report 2.

    Science.gov (United States)

    Leivo, Tiina; Sarikkola, Anna-Ulrika; Uusitalo, Risto J; Hellstedt, Timo; Ess, Sirje-Linda; Kivelä, Tero

    2011-06-01

    To present an economic-analysis comparison of simultaneous and sequential bilateral cataract surgery. Helsinki University Eye Hospital, Helsinki, Finland. Economic analysis. Effects were estimated from data in a study in which patients were randomized to have bilateral cataract surgery on the same day (study group) or sequentially (control group). The main clinical outcomes were corrected distance visual acuity, refraction, complications, Visual Function Index-7 (VF-7) scores, and patient-rated satisfaction with vision. Health-care costs of surgeries and preoperative and postoperative visits were estimated, including the cost of staff, equipment, material, floor space, overhead, and complications. The data were obtained from staff measurements, questionnaires, internal hospital records, and accountancy. Non-health-care costs of travel, home care, and time were estimated based on questionnaires from a random subset of patients. The main economic outcome measures were cost per VF-7 score unit change and cost per patient in simultaneous versus sequential surgery. The study comprised 520 patients (241 patients included non-health-care and time cost analyses). Surgical outcomes and patient satisfaction were similar in both groups. Simultaneous cataract surgery saved 449 Euros (€) per patient in health-care costs and €739 when travel and paid home-care costs were included. The savings added up to €849 per patient when the cost of lost working time was included. Compared with sequential bilateral cataract surgery, simultaneous bilateral cataract surgery provided comparable clinical outcomes with substantial savings in health-care and non-health-care-related costs. No author has a financial or proprietary interest in any material or method mentioned. Copyright © 2011 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  19. A toxicity study of simultaneous administration of Tamoxifen and Diazepam to female Wistar rats.

    Science.gov (United States)

    D'Mello, D; Mehta, D; Pereira, J; Rao, C V

    1999-11-01

    Tamoxifen (TAM) is used in the treatment of breast cancer and decreases the incidence of breast cancer when given to healthy women for different therapeutic purposes. This expansion of its use calls for further studies of its own potential side effects and those in combination with other prescription drugs. Diazepam (DP) is one such drug normally administered to patients who are under cancer treatment and those who suffer from insomnia. The present study examines the effect of individual and simultaneous administration of TAM and DP at therapeutic dose level of 0.8 mg/Kg/day of TAM and 0.3 mg/Kg/day of DP to normal female Wistar rats for a period of 12 weeks. The drugs were administered orally by mixing it in pellet made by wheat dough. There was no significant change in the terminal body weight and liver weights of animals. The ovary weights in TAM + DP treated animals were significantly decreased. The serum succinate dehydrogenase (SDH) levels were significantly lower in TAM, DP and TAM + DP treated rats and comparatively were lowest in TAM and TAM + DP treated animal groups. Serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT), acid and alkaline phosphatase (ACP & ALP) levels were significantly higher in the three treated groups, but comparatively lower in TAM + DP treated animals when compared to TAM or DP alone treated rats. There was marked increase in liver triglyceride and cholesterol levels in the three treated groups but remarkable decrease in liver glycogen. Total serum cholesterol levels were significantly high in DP and TAM + DP treated rats and total serum triglyceride levels were significantly high only in TAM treated rats. As a whole it can be concluded that DP does not enhance TAM toxicity on simultaneous administration, but on its own when administered individually brings about perturbation in lipid storage and metabolism.

  20. A device for simultaneous spin analysis of ultracold neutrons

    Energy Technology Data Exchange (ETDEWEB)

    Afach, S. [Institute for Particle Physics, ETH Zuerich, Zuerich (Switzerland); Paul Scherrer Institute, Villigen-PSI (Switzerland); Jena University Hospital, Hans Berger Department of Neurology, Jena (Germany); Ban, G.; Lefort, T.; Lemiere, Y.; Naviliat-Cuncic, O.; Quemener, G. [Universite de Caen, CNRS/IN2P3, LPC Caen ENSICAEN, Caen (France); Bison, G.; Chowdhuri, Z.; Daum, M.; Henneck, R.; Lauss, B.; Mtchedlishvili, A.; Schmidt-Wellenburg, P.; Zsigmond, G. [Paul Scherrer Institute, Villigen-PSI (Switzerland); Bodek, K.; Rawlik, M.; Rozpedzik, D.; Zejma, J. [Jagiellonian University, Marian Smoluchowski Institute of Physics, Cracow (Poland); Fertl, M.; Franke, B.; Kirch, K.; Komposch, S. [Institute for Particle Physics, ETH Zuerich, Zuerich (Switzerland); Paul Scherrer Institute, Villigen-PSI (Switzerland); Geltenbort, P. [Institut Laue-Langevin, Grenoble (France); Grujic, Z.D.; Kasprzak, M.; Weis, A. [University of Fribourg, Physics Department, Fribourg (Switzerland); Hayen, L.; Severijns, N.; Wursten, E. [Katholieke Universiteit Leuven, Instituut voor Kernen Stralingsfysica, Leuven (Belgium); Helaine, V. [Paul Scherrer Institute, Villigen-PSI (Switzerland); Universite de Caen, CNRS/IN2P3, LPC Caen ENSICAEN, Caen (France); Kermaidic, Y.; Pignol, G.; Rebreyend, D. [Universite Grenoble Alpes, CNRS/IN2P3, LPSC, Grenoble (France); Kozela, A. [Henryk Niedwodniczanski Institute for Nuclear Physics, Cracow (Poland); Krempel, J.; Piegsa, F.M. [Institute for Particle Physics, ETH Zuerich, Zuerich (Switzerland); Prashanth, P.N. [Paul Scherrer Institute, Villigen-PSI (Switzerland); Katholieke Universiteit Leuven, Instituut voor Kernen Stralingsfysica, Leuven (Belgium); Ries, D. [Paul Scherrer Institute, Villigen-PSI (Switzerland); Jena University Hospital, Hans Berger Department of Neurology, Jena (Germany); Roccia, S. [Universite Paris Sud, CNRS/IN2P3, CSNSM, Orsay campus (France); Wyszynski, G. [Institute for Particle Physics, ETH Zuerich, Zuerich (Switzerland); Jagiellonian University, Marian Smoluchowski Institute of Physics, Cracow (Poland)

    2015-11-15

    We report on the design and first tests of a device allowing for measurement of ultracold neutrons polarisation by means of the simultaneous analysis of the two spin components. The device was developed in the framework of the neutron electric dipole moment experiment at the Paul Scherrer Institute. Individual parts and the entire newly built system have been characterised with ultracold neutrons. The gain in statistical sensitivity obtained with the simultaneous spin analyser is (18.2 ± 6.1) % relative to the former sequential analyser under nominal running conditions. (orig.)

  1. Simultaneous pulmonary and systemic blood pressure and ECG Interval measurement in conscious, freely moving rats.

    Science.gov (United States)

    Rey, Markus; Weber, Edgar W; Hess, Patrick D

    2012-03-01

    Here we evaluated the ability of a new, dual blood-pressure telemetry transmitter to simultaneously measure pulmonary and systemic blood pressure and the electrocardiogram in rats. The transmitter was implanted in normotensive and monocrotaline-induced pulmonary hypertensive Wistar rats, with sensing catheters placed in the pulmonary artery (channel 1) and descending aorta (channel 2). Biopotential electrodes were positioned to record an apex-based lead II electrocardiogram. Pulmonary and systemic arterial blood pressure and electrocardiographic waveforms were recorded between 2 and 12 wk after implantation of the transmitter. During this period, pulmonary arterial pressure progressively increased in monocrotaline-treated compared with saline-treated rats. The pharmacologic response of rats to reference compounds was measured by using the transmitter to validate the technique and to evaluate the ability of the device to transmit changes in blood pressure and the electrocardiogram. Validation against 2 Millar high-fidelity blood-pressure catheters confirmed the accuracy of the blood pressure data recorded with the transmitter. In addition, local tolerance of the associated catheters was confirmed by histologic examination.

  2. Simultaneous determination of mangiferin and neomangiferin in rat plasma by UPLC-MS/MS and its application for pharmacokinetic study.

    Science.gov (United States)

    Qiu, Xiangjun; Zhao, Jian-long; Hao, Cong; Yuan, Canli; Tian, Nuan; Xu, Zhi-sheng; Zou, Ruan-min

    2016-05-30

    In this study, a sensitive and rapid ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine mangiferin and neomangiferin in rat plasma simultaneously. Chromatographic separation was carried out on an Acquity UPLC BEH C18 column and mass spectrometric analysis was performed using a Xevo TQD triple quadruple mass spectrometer coupled with an electrospray ionization (ESI) source. The MRM transitions of m/z 423.2 → 303.1 and m/z 585.0 → 273.1 were used to quantify for mangiferin and neomangiferin, respectively. The linearity of this method was found to be within the concentration range of 5-2000 ng/mL for mangiferin, and 2-1000 ng/mL for neomangiferin in rat plasma, respectively. Only 3.0 min was needed for an analytical run. This assay was used to support a preclinical study to investigate the pharmacokinetics of mangiferin and neomangiferin in rats.

  3. Effects of simultaneous exposure to stress and nicotine on nicotine-induced locomotor activation in adolescent and adult rats

    Energy Technology Data Exchange (ETDEWEB)

    Zago, A. [Laboratório de Farmacologia, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara, SP (Brazil); Leão, R.M.; Carneiro-de-Oliveira, P.E. [Laboratório de Farmacologia, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara, SP (Brazil); Programa Interinstitucional de Pós-Graduação em Ciências Fisiológicas, Universidade Federal de São Carlos/Universidade Estadual de São Paulo, Araraquara, SP (Brazil); Marin, M.T.; Cruz, F.C. [Laboratório de Farmacologia, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara, SP (Brazil); Planeta, C.S. [Laboratório de Farmacologia, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara, SP (Brazil); Programa Interinstitucional de Pós-Graduação em Ciências Fisiológicas, Universidade Federal de São Carlos/Universidade Estadual de São Paulo, Araraquara, SP (Brazil)

    2011-11-18

    Preclinical studies have shown that repeated stress experiences can result in an increase in the locomotor response to the subsequent administration of drugs of abuse, a phenomenon that has been termed behavioral cross-sensitization. Behavioral sensitization reflects neuroadaptive processes associated with drug addiction and drug-induced psychosis. Although crosssensitization between stress- and drug-induced locomotor activity has been clearly demonstrated in adult rats, few studies have evaluated this phenomenon in adolescent rats. In the present study, we determined if the simultaneous exposure to stress and nicotine was capable of inducing behavioral sensitization to nicotine in adolescent and adult rats. To this end, adolescent (postnatal day (P) 28-37) and adult (P60-67) rats received nicotine (0.4 mg/kg, sc) or saline (0.9% NaCl, sc) and were immediately subjected to restraint stress for 2 h once a day for 7 days. The control group for stress was undisturbed following nicotine or saline injections. Three days after the last exposure to stress and nicotine, rats were challenged with a single dose of nicotine (0.4 mg/kg, sc) or saline and nicotine-induced locomotion was then recorded for 30 min. In adolescent rats, nicotine caused behavioral sensitization only in animals that were simultaneously exposed to stress, while in adult rats nicotine promoted sensitization independently of stress exposure. These findings demonstrate that adolescent rats are more vulnerable to the effects of stress on behavioral sensitization to nicotine than adult rats.

  4. High Performance Liquid Chromatographic Assay for the Simultaneous Determination of Posaconazole and Vincristine in Rat Plasma

    Directory of Open Access Journals (Sweden)

    Hadeel A. Khalil

    2015-01-01

    Full Text Available Purpose. Developing a validated HPLC-DAD method for simultaneous determination of posaconazole (PSZ and vincristine (VCR in rat plasma. Methods. PSZ, VCR, and itraconazole (ITZ were extracted from 200 μL plasma using diethyl ether in the presence of 0.1 M sodium hydroxide solution. The organic layer was evaporated in vacuo and dried residue was reconstituted and injected through HC-C18 (4.6 × 250 mm, 5 μm column. In the mobile phase, acetonitrile and 0.015 M potassium dihydrogen orthophosphate (30 : 70 to 80 : 20, linear gradient over 7 minutes pumped at 1.5 mL/min. VCR and PSZ were measured at 220 and 262 nm, respectively. Two Sprague Dawley rats were orally dosed PSZ followed by iv dosing of VCR and serial blood sampling was performed. Results. VCR, PSZ, and ITZ were successfully separated within 11 min. Calibration curves were linear over the range of 50–5000 ng/mL for both drugs. The CV% and % error of the mean were ≤18% and limit of quantitation was 50 ng/mL for both drugs. Rat plasma concentrations of PSZ and VCR were simultaneously measured up to 72 h and their calculated pharmacokinetics parameters were comparable to the literature. Conclusion. The assay was validated as per ICH guidelines and is appropriate for pharmacokinetics drug-drug interaction studies.

  5. The effects of simultaneous combined exposure to CDMA and WCDMA electromagnetic fields on rat testicular function.

    Science.gov (United States)

    Lee, Hae-June; Jin, Yeung Bae; Kim, Tae-Hong; Pack, Jeong-Ki; Kim, Nam; Choi, Hyung-Do; Lee, Jae-Seon; Lee, Yun-Sil

    2012-05-01

    Wireless mobile phones and other telecommunication devices are used extensively in daily life. We therefore examined the effects of combined exposure to radiofrequency electromagnetic fields (RF-EMF) on rat testicular function, specifically with respect to sensitive processes such as spermatogenesis. Male rats were exposed to single code division multiple access (CDMA) and wideband code division multiple access (WCDMA) RF signals for 12 weeks. The RF exposure schedule comprised 45 min/day, 5 days/week for a total of 12 weeks. The whole-body average specific absorption rate (SAR) of CDMA and WCDMA was 2.0 W/kg each or 4.0 W/kg in total. We then investigated the correlates of testicular function such as sperm count in the cauda epididymis, testosterone concentration in the blood serum, malondialdehyde concentrations in the testes and epididymis, frequency of spermatogenesis stages, and appearance of apoptotic cells in the testes. We also immunoblotted for p53, bcl2, GADD45, cyclin G, and HSP70 in the testes of sham- and combined RF-exposed animals. Based on the results, we concluded that simultaneous exposure to CDMA and WCDMA RF-EMFs at 4.0 W/kg SAR did not have any observable adverse effects on rat spermatogenesis.

  6. Comprehensive model for simultaneous MRI determination of perfusion and permeability using a blood-pool agent in rats rhabdomyosarcoma

    Energy Technology Data Exchange (ETDEWEB)

    Bazelaire, Cedric de [Saint Louis Hospital, Radiology Department, Paris (France); Siauve, Nathalie; Fournier, Laure; Clement, Olivier; Kerviler, Eric de; Cuenod, Charles Andre [George Pompidou European Hospital, Radiology Department, Paris (France); Frouin, Frederique [INSERM U494, Faculte de Medecine Pitie-Salpetriere, Paris (France); Robert, Philippe [Guerbet Laboratoire Guerbet, Recherche et Developpement, Paris (France)

    2005-12-01

    To present a new compartmental analysis model developed to simultaneously measure tissue perfusion and capillary permeability in a tumor using MRI and a macromolecular contrast medium. Rhadomyosarcomas were implanted subcutaneously in 20 rats and studied by 1.5-T MRI using a fast gradient echo sequence (2D fast SPGR TR/TE/{alpha} 13 ms/1.2 ms/60 ) after injection of a macromolecular contrast medium. The left ventricle and tumor signal intensities were converted into concentrations and modeled using compartmental analysis, yielding tumor perfusion F, distribution volume V{sub distribution}, volume transfer constant K{sup trans}, rate constant of influx k{sub pe}, and initial extraction (fraction) E. Tumor perfusion was F=43{+-}29 ml.min{sup -1}.100 g{sup -1}. The permeability study allowed the measurement of k{sub pe}=0.37{+-}0.12 min{sup -1} and K{sup trans}=0.01{+-}0.0031 min{sup -1}. The blood volume could be assimilated to the distribution volume (V{sub distribution}=2.9{+-}1.01%) since the capillary leakage was small. The simultaneous assessment of perfusion and permeability allowed quantification of the initial extraction (fraction) E=2.34{+-}1.05%. Quantification of both tumor perfusion and capillary leakage is feasible using MRI using a macromolecular blood pool agent. The method should improve tumor characterization. (orig.)

  7. Simultaneous Determination of Silybin A and Silybin B in Rat Plasma and Pharmacokinetic Study

    Institute of Scientific and Technical Information of China (English)

    CHU Yang; LI Wei; LI Zhi-wen; LI Xin-xin; MA Xiao-hui; ZHOU Shui-ping; ZHU Yong-hong

    2011-01-01

    Objective To investigate the bioavailability and pharmacokinetics of silybin A and silybin B in rats,respectively.Methods Following iv and ig administration of silybin to 20 Wistar rats,the plasma samples were collected at different time points up to 12 h.Sample pretreatment was involved in one-step protein precipitation with acetonitrile.Silybin A and silybin B were simultaneously determined by LC-MS/MS.Results After ig dosing silybin 28,56,and 112 mg/kg to rats,the t1/2β values were 5.48,5.08,and 5.73 h for silybin A,and 4.56,4.12,and 5.53 h for silybin B; The Cmax were 674.3,1349.4,and 2042.5 ng/mL for silybin A,and 671.0,1365.4,and 2066.2 ng/mL for silybin B; The Tmax were 0.20,0.23,and 0.20 h for silybin A,and 0.20,0.23,and 0.20 h for silybin B; The AUC were 454.4,845.9,and 1219.5 h·ng/mL for silybin A,and 432.0,817.1,and 1153.6 h·ng/mL for silybin B.The absolute bioavailabilities of silybin A and silybin B were 2.86% and 1.93%,respectively.Conclusion Silybin A and silybin B have very low bioavailability after ig administration,and there is no significant difference in the pharmacokinetic parameters between silybin A and silybin B,which indicates that the two diastereoisomers have similar pharmacokinetic behavior in rats.

  8. HPLC method for simultaneous determination of clobazam and N-desmethylclobazam in human serum, rat serum and rat brain homogenates.

    Science.gov (United States)

    Gaździk, W R; Podleśny, J; Filipek, M

    1989-03-01

    A method for simultaneous determination of clobazam (CBZ) and its active metabolite N-desmethylclobazam (DCBZ) in various biological samples by RP-HPLC with UV detection is described. The determination of both CBZ and DCBZ is performed without derivatization. The internal standard is diazepam. The method is rapid and simple with sensitivity limits of 10 ng/mL for both CBZ and DCBZ and is suitable for routine analysis as well as for animal studies.

  9. Simultaneous determination of three polyphenols in rat plasma after orally administering hawthorn leaves extract by the HPLC method.

    Science.gov (United States)

    Ying, Xixiang; Meng, Xiansheng; Wang, Siyuan; Wang, Dong; Li, Haibo; Wang, Bing; Du, Yang; Liu, Xun; Zhang, Wenjie; Kang, Tingguo

    2012-01-01

    A simple and sensitive HPLC method was developed to simultaneously determine three active compounds, vitexin-4″-O-glucoside (VG), vitexin-2″-O-rhamnoside (VR) and hyperoside (HP), in rat plasma after administering the hawthorn leaves extract (HLE). An HPLC assay with baicalin as the internal standard was carried out using a Phenomsil C₁₈ analytical column with UV detection at 332 nm. The mobile phase consisted of methanol-acetonitrile-tetrahydrofuran-1% glacial acetic acid (6 : 1.5 : 18.5 : 74, v/v/v/v). The calibration curves were linear over the range of 2.5-500, 0.2-25 and 0.25-12.5 µg mL⁻¹ for VG, VR and HP, respectively. The method was reproducible and reliable, with relative standard deviations of the intra- and inter-day precision between 1.2% and 13.2% for the analysis of the three analytes. The validated HPLC method herein described was successfully applied to the pharmacokinetic study of VG, VR and HP after oral administration of HLE to rats over the dose range of 2.5-10  mL kg⁻¹.

  10. Simultaneous quantification of two canthinone alkaloids of Picrasma quassioides in rat plasma by liquid chromatography-tandem mass spectrometry and its application to a rat pharmacokinetic study.

    Science.gov (United States)

    Shi, Yuanyuan; Hong, Chunyan; Xu, Jian; Yang, Xiaoling; Xie, Ning; Feng, Feng; Liu, Wenyuan

    2015-04-01

    Picrasma quassioides (D. Don) Benn. is used in traditional Chinese medicine for the treatment of inflammation. Characteristic components of the medicinal extract are canthinone alkaloids. In this study, a sensitive and rapid liquid chromatography with tandem mass spectrometry method has been developed for simultaneous quantification of two major canthinone alkaloids, 5-hydroxy-4-methoxycanthin-6-one and 4,5-dimethoxycanthin-6-one, in rat plasma after oral administration of P. quassioides extract (200 mg/kg). The chromatographic separation was performed on a C18 column using acetonitrile-aqueous 0.1% formic acid (90:10, v/v) as the mobile phase. Plasma samples were prepared for analysis using a simple liquid-liquid extraction with ethyl acetate. Analytes were detected using tandem mass spectrometry in positive multiple reaction monitoring mode. Method validation revealed excellent linearity over the range 1.25-900 ng/mL for 5-hydroxy-4-methoxycanthin-6-one and 0.5-800 ng/mL for 4,5-dimethoxycanthin-6-one with satisfactory intra- and inter-day precision, accuracy and recovery. Samples were stable under the conditions tested. The pharmacokinetic profiles of the analytes in rats showed that both canthinones were rapidly absorbed and that 4,5-dimethoxycanthin-6-one was eliminated faster than 5-hydroxy-4-methoxycanthin-6-one.

  11. Integrative Data Analysis: The Simultaneous Analysis of Multiple Data Sets

    Science.gov (United States)

    Curran, Patrick J.; Hussong, Andrea M.

    2009-01-01

    There are both quantitative and methodological techniques that foster the development and maintenance of a cumulative knowledge base within the psychological sciences. Most noteworthy of these techniques is meta-analysis, which allows for the synthesis of summary statistics drawn from multiple studies when the original data are not available.…

  12. SIMULTANEOUS MEASUREMENT OF EXTRACELLULAR MORPHINE AND SEROTONIN IN BRAIN-TISSUE AND CSF BY MICRODIALYSIS IN AWAKE RATS

    NARCIS (Netherlands)

    MATOS, FF; ROLLEMA, H; BASBAUM, AI

    1992-01-01

    In this report, we describe an HPLC with electrochemical detection assay for the simultaneous measurement of levels of morphine, serotonin, 5-hydroxyindole-3-acetic acid, and homovanillic acid in dialysates of various brain areas and CSF in the awake rat. Morphine could be detected in the dialysates

  13. Non-chemosensitive parafacial neurons simultaneously regulate active expiration and airway patency under hypercapnia in rats.

    Science.gov (United States)

    de Britto, Alan A; Moraes, Davi J A

    2017-03-15

    Hypercapnia or parafacial respiratory group (pFRG) disinhibition at normocapnia evokes active expiration in rats by recruitment of pFRG late-expiratory (late-E) neurons. We show that hypercapnia simultaneously evoked active expiration and exaggerated glottal dilatation by late-E synaptic excitation of abdominal, hypoglossal and laryngeal motoneurons. Simultaneous rhythmic expiratory activity in previously silent pFRG late-E neurons, which did not express the marker of ventral medullary CO2 -sensitive neurons (transcription factor Phox2b), was also evoked by hypercapnia. Hypercapnia-evoked active expiration, neural and neuronal late-E activities were eliminated by pFRG inhibition, but not after blockade of synaptic excitation. Hypercapnia produces disinhibition of non-chemosensitive pFRG late-E neurons to evoke active expiration and concomitant cranial motor respiratory responses controlling the oropharyngeal and upper airway patency. Hypercapnia produces active expiration in rats and the recruitment of late-expiratory (late-E) neurons located in the parafacial respiratory group (pFRG) of the ventral medullary brainstem. We tested the hypothesis that hypercapnia produces active expiration and concomitant cranial respiratory motor responses controlling the oropharyngeal and upper airway patency by disinhibition of pFRG late-E neurons, but not via synaptic excitation. Phrenic nerve, abdominal nerve (AbN), cranial respiratory motor nerves, subglottal pressure, and medullary and spinal neurons/motoneurons were recorded in in situ preparations of juvenile rats. Hypercapnia evoked AbN active expiration, exaggerated late-E discharges in cranial respiratory motor outflows, and glottal dilatation via late-E synaptic excitation of abdominal, hypoglossal and laryngeal motoneurons. Simultaneous rhythmic late-E activity in previously silent pFRG neurons, which did not express the marker of ventral medullary CO2 -sensitive neurons (transcription factor Phox2b), was also evoked by

  14. Truss topology optimization with simultaneous analysis and design

    Science.gov (United States)

    Sankaranarayanan, S.; Haftka, Raphael T.; Kapania, Rakesh K.

    1992-01-01

    Strategies for topology optimization of trusses for minimum weight subject to stress and displacement constraints by Simultaneous Analysis and Design (SAND) are considered. The ground structure approach is used. A penalty function formulation of SAND is compared with an augmented Lagrangian formulation. The efficiency of SAND in handling combinations of general constraints is tested. A strategy for obtaining an optimal topology by minimizing the compliance of the truss is compared with a direct weight minimization solution to satisfy stress and displacement constraints. It is shown that for some problems, starting from the ground structure and using SAND is better than starting from a minimum compliance topology design and optimizing only the cross sections for minimum weight under stress and displacement constraints. A member elimination strategy to save CPU time is discussed.

  15. Effects of simultaneous exposure to stress and nicotine on nicotine-induced locomotor activation in adolescent and adult rats

    Directory of Open Access Journals (Sweden)

    A. Zago

    2012-01-01

    Full Text Available Preclinical studies have shown that repeated stress experiences can result in an increase in the locomotor response to the subsequent administration of drugs of abuse, a phenomenon that has been termed behavioral cross-sensitization. Behavioral sensitization reflects neuroadaptive processes associated with drug addiction and drug-induced psychosis. Although cross-sensitization between stress- and drug-induced locomotor activity has been clearly demonstrated in adult rats, few studies have evaluated this phenomenon in adolescent rats. In the present study, we determined if the simultaneous exposure to stress and nicotine was capable of inducing behavioral sensitization to nicotine in adolescent and adult rats. To this end, adolescent (postnatal day (P 28-37 and adult (P60-67 rats received nicotine (0.4 mg/kg, sc or saline (0.9% NaCl, sc and were immediately subjected to restraint stress for 2 h once a day for 7 days. The control group for stress was undisturbed following nicotine or saline injections. Three days after the last exposure to stress and nicotine, rats were challenged with a single dose of nicotine (0.4 mg/kg, sc or saline and nicotine-induced locomotion was then recorded for 30 min. In adolescent rats, nicotine caused behavioral sensitization only in animals that were simultaneously exposed to stress, while in adult rats nicotine promoted sensitization independently of stress exposure. These findings demonstrate that adolescent rats are more vulnerable to the effects of stress on behavioral sensitization to nicotine than adult rats.

  16. Oleuropein improves glucose tolerance and lipid profile in rats with simultaneous renovascular hypertension and type 2 diabetes.

    Science.gov (United States)

    Khalili, Azadeh; Nekooeian, Ali Akbar; Khosravi, Mohammad Bagher

    2017-10-01

    Oleuropein mediates most of the beneficial effects of olive products. This study examined the role of oxidative stress in the effects of oleuropein on lipid profile and blood glucose in rats with simultaneous renovascular hypertension and type 2 diabetes. Eight groups (n = 7-9 each) of male Sprague-Dawley rats including a control, a type 2 diabetic, a renovascular hypertensive, a sham, a simultaneously hypertensive diabetic receiving vehicle, and 3 simultaneously hypertensive-diabetic receiving 20, 40, or 60 mg/kg/day oleuropein were used. Four weeks after treatment, blood glucose, lipid profile, and biomarkers of oxidative stress were measured, and glucose tolerance test (GTT) was performed. Simultaneously hypertensive diabetic rats had significantly higher blood pressure, blood glucose, and serum total cholesterol, low-density lipoprotein cholesterol (LDL-C), triglyceride and malondialdehyde. They also had lower serum high-density lipoprotein cholesterol, erythrocyte superoxide dismutase, and impaired glucose tolerance. Oleuropein significantly reduced blood pressure, blood glucose, and serum total cholesterol, LDL-C, triglyceride and malondoaldehyde. It also increased serum high-density lipoprotein cholesterol, erythrocyte superoxide dismutase, and improved glucose tolerance. The findings show that the model is associated with impaired glucose tolerance, and adverse lipid profile. They also show that oleuropein, partly by an antioxidant mechanism, improves glucose tolerance and changed lipid profile favorably.

  17. Simultaneous thermal analysis and thermodilatometry of hybrid fiber reinforced UHPC

    Science.gov (United States)

    Scheinherrová, Lenka; Fořt, Jan; Pavlík, Zbyšek; Černý, Robert

    2017-07-01

    Development of concrete technology and the availability of variety of materials such as silica fume, mineral microfillers and high-range water-reducing admixtures make possible to produce Ultra-High Performance Concrete (UHPC) with compressive strength higher than 160 MPa. However, UHPC is prone to spall under high temperatures what limits its use for special applications only, such as offshore and marine structures, industrial floors, security barriers etc. The spalling is caused by the thermal stresses due to the temperature gradient during heating, and by the splitting force owing to the release of water vapour. Hybrid fibre reinforcement based on combination of steel and polymer fibres is generally accepted by concrete community as a functional solution preventing spalling. In this way, Ultra-High Performance Fibre Reinforced Concrete (UHPFRC) is produced possessing high mechanical strength, durability and resistance to water and salt ingress. Since UHPFRC find use in construction industry in tunnel linings, precast tunnel segments, and high-rise buildings, its behaviour during the high-temperature exposure and its residual parameters are of the particular importance. On this account, Simultaneous Thermal Analysis (STA) and Thermodilatometry Analysis (TDA) were done in the paper to identify the structural and chemical changes in UHPFRC during its high-temperature load. Based on the experimental results, several physical and chemical processes that studied material underwent at high-temperatures were recognized. The obtained data revealed changes in the composition of the studied material and allowed identification of critical temperatures for material damage.

  18. Simultaneous Thermal Analysis of Remediated Nitrate Salt Surrogates

    Energy Technology Data Exchange (ETDEWEB)

    Wayne, David Matthew [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2016-05-13

    The actinide engineering and science group (MET-1) have completed simultaneous thermal analysis and offgas analysis by mass spectrometry (STA-MS) of remediated nitrate salt (RNS) surrogates formulated by the high explosives science and technology group (M-7). The 1.0 to 1.5g surrogate samples were first analyzed as received, then a new set was analyzed with 100-200mL 10M HNO3 +0.3 MHF added, and a third set was analyzed after 200 mL of a concentrated Pu-AM spike (in 10M HNO3 +0.3 MHF) was added. The acid and spike solutions were formulated by the actinide analytical chemistry group (C-AAC) using reagent-grade HNO3 and HF, which was also used to dissolve a small quantity of mixed, high-fired PuO2/ AmO2 oxide.

  19. UHPFRC at high temperatures - Simultaneous thermal analysis and thermodilatometry

    Science.gov (United States)

    Trník, Anton; Fořt, Jan; Pavlíková, Milena; Čáchová, Monika; Čítek, David; Kolísko, Jiří; Černý, Robert; Pavlík, Zbyšek

    2016-07-01

    Simultaneous Thermal Analysis (STA) and Thermodilatometry Analysis (TDA) are done to reveal the structural and chemical changes in UHPFRC during its high-temperature load. Based on the measured results, several physical and chemical processes that studied material underwent at high-temperatures are recognized. In the temperature interval from 25 to 300 °C, the liberation of physically bound water from pores and the dehydration reaction of C-S-H take place. Additionally, AFt and AFm phases dehydrate at 110 - 156 °C. Endothermic peat at 460 °C corresponds to the portlandite decomposition. At 575 °C, the α → β transformation of quartz is found. This reaction is accompanied by a sharp endothermic heat flow peak and a volume expansion, whereas no change of mass is measured. In the temperature interval 580-800 °C, the calcite and C-S-H gels decomposition is monitored. At the temperature above 800 °C, there is one significant exothermal peak corresponding to a crystallization of wollastonite. In summary, STA and TDA data pointed out the structural changes in studied UHPFRC and allowed identification of critical temperatures for its damage.

  20. A Validated HPLC-DAD Method for Simultaneous Determination of Etodolac and Pantoprazole in Rat Plasma

    Directory of Open Access Journals (Sweden)

    Ali S. Abdelhameed

    2014-01-01

    Full Text Available A simple, sensitive, and accurate HPLC-DAD method has been developed and validated for the simultaneous determination of pantoprazole and etodolac in rat plasma as a tool for therapeutic drug monitoring. Optimal chromatographic separation of the analytes was achieved on a Waters Symmetry C18 column using a mobile phase that consisted of phosphate buffer pH~4.0 as eluent A and acetonitrile as eluent B in a ratio of A : B, 55 : 45 v/v for 6 min, pumped isocratically at a flow rate of 0.8 mL min−1. The eluted analytes were monitored using photodiode array detector set to quantify samples at 254 nm. The method was linear with r2=0.9999 for PTZ and r2=0.9995 for ETD at a concentration range of 0.1–15 and 5–50 μgmL−1 for PTZ and ETD, respectively. The limits of detection were found to be 0.033 and 0.918 μgmL−1 for PTZ and ETD, respectively. The method was statistically validated for linearity, accuracy, precision, and selectivity following the International Conference for Harmonization (ICH guidelines. The reproducibility of the method was reliable with the intra- and interday precision (% RSD <7.76% for PTZ and <7.58 % for ETD.

  1. Effects of simultaneous combined exposure to CDMA and WCDMA electromagnetic fields on serum hormone levels in rats.

    Science.gov (United States)

    Jin, Yeung Bae; Choi, Hyung-Do; Kim, Byung Chan; Pack, Jeong-Ki; Kim, Nam; Lee, Yun-Sil

    2013-05-01

    Despite more than a decade of research on the endocrine system, there have been no published studies about the effects of concurrent exposure of radiofrequency electromagnetic fields (RF-EMF) on this system. The present study investigated the several parameters of the endocrine system including melatonin, thyroid stimulating hormone, stress hormone and sex hormone after code division multiple access (CDMA, 849 MHz) and wideband code division multiple access (WCDMA, 1.95 GHz) signals for simultaneous exposure in rats. Sprague-Dawley rats were exposed to RF-EMF signals for 45 min/day, 5 days/week for up to 8 weeks. The whole-body average specific absorption rate (SAR) of CDMA or WCDMA was 2.0 W/kg (total 4.0 W/kg). At 4 and 8 weeks after the experiment began, each experimental group's 40 rats (male 20, female 20) were autopsied. Exposure for 8 weeks to simultaneous CDMA and WCDMA RF did not affect serum levels in rats of melatonin, thyroid stimulating hormone (TSH), triiodothyronine (T3) and thyroxin (T4), adrenocorticotropic hormone (ACTH) and sex hormones (testosterone and estrogen) as assessed by the ELISA method.

  2. Rat sperm motility analysis: methodologic considerations

    Science.gov (United States)

    The objective of these studies was to optimize conditions for computer-assisted sperm analysis (CASA) of rat epididymal spermatozoa. Methodologic issues addressed include sample collection technique, sampling region within the epididymis, type of diluent medium used, and sample c...

  3. Simultaneous comprehensive multiplex autoantibody analysis for rapidly progressive glomerulonephritis.

    Science.gov (United States)

    Sowa, Mandy; Trezzi, Barbara; Hiemann, Rico; Schierack, Peter; Grossmann, Kai; Scholz, Juliane; Somma, Valentina; Sinico, Renato Alberto; Roggenbuck, Dirk; Radice, Antonella

    2016-11-01

    Rapidly progressive glomerulonephritis (RPGN) is mainly caused by anti-glomerular basement membrane (GBM) antibody-mediated glomerulonephritis, immune-complex or anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides and leads to rapid loss of renal function. Detection of ANCA and autoantibodies (autoAbs) to GBM and dsDNA enables early diagnosis and appropriate treatment of RPGN aiding in preventing end-stage renal disease.Determination of ANCA on neutrophils (ANCA) as well as autoAbs to myeloperoxidase (MPO-ANCA), proteinase 3 (PR3-ANCA), GBM, and dsDNA was performed by the novel multiplex CytoBead technology combining cell- and microbead-based autoAb analyses by automated indirect immunofluorescence (IIF). Forty patients with granulomatosis with polyangiitis (GPA), 48 with microscopic polyangiitis (MPA), 2 with eosinophilic GPA, 42 with systemic lupus erythematosus (SLE), 43 with Goodpasture syndrome (GPS), 57 with infectious diseases (INF), and 55 healthy subjects (HS) were analyzed and findings compared with classical single testing.The CytoBead assay revealed for GPA, MPA, GPS, and SLE the following diagnostic sensitivities and for HS and INF the corresponding specificities: PR3-ANCA, 85.0% and 100.0%; MPO-ANCA, 77.1% and 99.1%; anti-GBM autoAb, 88.4% and 96.4%; anti-dsDNA autoAb, 83.3% and 97.3%; ANCA, 91.1% and 99.1%, respectively. Agreement with classical enzyme-linked immunosorbent assay and IIF was very good for anti-GBM autoAb, MPO-ANCA, PR3-ANCA, and ANCA, respectively. Anti-dsDNA autoAb comparative analysis demonstrated fair agreement only and a significant difference (P = 0.0001).The CytoBead technology provides a unique multiplex reaction environment for simultaneous RPGN-specific autoAb testing. CytoBead RPGN assay is a promising alternative to time-consuming single parameter analysis and, thus, is well suited for emergency situations.

  4. Simultaneous comprehensive multiplex autoantibody analysis for rapidly progressive glomerulonephritis

    Science.gov (United States)

    Sowa, Mandy; Trezzi, Barbara; Hiemann, Rico; Schierack, Peter; Grossmann, Kai; Scholz, Juliane; Somma, Valentina; Sinico, Renato Alberto; Roggenbuck, Dirk; Radice, Antonella

    2016-01-01

    Abstract Rapidly progressive glomerulonephritis (RPGN) is mainly caused by anti-glomerular basement membrane (GBM) antibody-mediated glomerulonephritis, immune-complex or anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides and leads to rapid loss of renal function. Detection of ANCA and autoantibodies (autoAbs) to GBM and dsDNA enables early diagnosis and appropriate treatment of RPGN aiding in preventing end-stage renal disease. Determination of ANCA on neutrophils (ANCA) as well as autoAbs to myeloperoxidase (MPO-ANCA), proteinase 3 (PR3-ANCA), GBM, and dsDNA was performed by the novel multiplex CytoBead technology combining cell- and microbead-based autoAb analyses by automated indirect immunofluorescence (IIF). Forty patients with granulomatosis with polyangiitis (GPA), 48 with microscopic polyangiitis (MPA), 2 with eosinophilic GPA, 42 with systemic lupus erythematosus (SLE), 43 with Goodpasture syndrome (GPS), 57 with infectious diseases (INF), and 55 healthy subjects (HS) were analyzed and findings compared with classical single testing. The CytoBead assay revealed for GPA, MPA, GPS, and SLE the following diagnostic sensitivities and for HS and INF the corresponding specificities: PR3-ANCA, 85.0% and 100.0%; MPO-ANCA, 77.1% and 99.1%; anti-GBM autoAb, 88.4% and 96.4%; anti-dsDNA autoAb, 83.3% and 97.3%; ANCA, 91.1% and 99.1%, respectively. Agreement with classical enzyme-linked immunosorbent assay and IIF was very good for anti-GBM autoAb, MPO-ANCA, PR3-ANCA, and ANCA, respectively. Anti-dsDNA autoAb comparative analysis demonstrated fair agreement only and a significant difference (P = 0.0001). The CytoBead technology provides a unique multiplex reaction environment for simultaneous RPGN-specific autoAb testing. CytoBead RPGN assay is a promising alternative to time-consuming single parameter analysis and, thus, is well suited for emergency situations. PMID:27858870

  5. ANOVA-principal component analysis and ANOVA-simultaneous component analysis: a comparison.

    NARCIS (Netherlands)

    Zwanenburg, G.; Hoefsloot, H.C.J.; Westerhuis, J.A.; Jansen, J.J.; Smilde, A.K.

    2011-01-01

    ANOVA-simultaneous component analysis (ASCA) is a recently developed tool to analyze multivariate data. In this paper, we enhance the explorative capability of ASCA by introducing a projection of the observations on the principal component subspace to visualize the variation among the measurements.

  6. Immunohistochemistry Study on Androgen and Estrogen Receptors of Rat Seminal Vesicle Submitted to Simultaneous Alcohol-Nicotine Treatment

    Directory of Open Access Journals (Sweden)

    Mohsen Basiri

    2016-09-01

    Full Text Available Objective: Alcohol consumption is habitually accompanied by the use of other psychoactive substances, mostly tobacco. Nicotine and alcohol affect male accessory reproductive glands function. Most studies have been done on pathologic features of prostate, but there has been no systematic study on the seminal vesicle. Therefore, the aim of current study was to investigate the distribution of androgen receptor (AR and estrogen receptors-beta (ER-β immune reactivities following long-term treatment of alcohol, nicotine or a combination of both substances. Materials and Methods: In this experimental study, a total of 40 adult Wistar rats, nine weeks of age, were used. Animals were randomly divided into four groups, including: i. Control group receiving normal saline 0.09%, ii. Ethanol group receiving ethanol 20% (2 ml/kg, via gavage, iii. Nicotine group receiving nicotine (0.1 mg/kg, subcutaneous injection, and iv. Ethanol-nicotine group receiving simultaneous ethanol 20% (2 ml/kg and nicotine (0.1 mg/kg treatment. All treatment lasted for eight weeks. Prior to intracardiac perfusion, blood sample was collected from left ventricle. The seminal vesicles were isolated and processed for paraffin blocking. The sample tissues were then studied for distribution of AR and ER-β immunereactivities using immunohistochemical (IHC staining method. One way analysis of variance (ANOVA and Tukey’s test were performed for data analysis. A value of P<0.05 was considered significant. Results: Our results revealed that the lowest mean number of positive cells belonged to the animals of ethanol-nicotine group that was followed by the ethanol, nicotine, and control groups, respectively. However, there was no significant difference regarding serum testosterone level among experimental groups. Conclusion: It was concluded that combination of both ethanol and nicotine may be a crucial factor in the expression levels of AR and ER-β.

  7. Simultaneous-equations Analysis in Regional Science and Economic Geography

    DEFF Research Database (Denmark)

    Mitze, Timo; Stephan, Andreas

    This paper provides an overview over simultaneous equation models (SEM) in the context of analyses based on regional data. We describe various modelling approaches and highlight close link of SEMs to theory and also comment on the advantages and disadvantages of SEMs.We present selected empirical...

  8. Simultaneous quantification of picfeltarraenins IA and IB in rat plasma by UPLC-MS/MS: Application to a pharmacokinetic study.

    Science.gov (United States)

    He, Xin; Zhang, Yingjie; Gao, Hang; Li, Keyan; Zhang, Yazhuo; Sun, Limin; Tao, Guizhou

    2016-02-20

    A simple and rapid quantitative UPLC-MS/MS method for simultaneous determination of picfeltarraenins IA and IB in rat plasma was developed and validated in accordance with the US FDA Bioanalytical Guidance (2001). Analytes were extracted from rat plasma by using methanol and separated on Agilent ZORBAX SB-C18 (50mm×2.1mm, 1.8μm) column by using a mobile phase composed of methanol and water (70:30, v/v). Eluents were monitored by ESI tandem mass spectrometry detection with SRM mode using ion transitions m/z 785.4→639.5, m/z 815.5→669.5, and m/z 763.5→455.3 for picfeltarraenin IA, picfeltarraenin IB, and internal standard, respectively. The method was validated over the linear range of 11.5-1150ng/mL and 13.0-1300ng/mL. The developed analytical method was applied to support a pharmacokinetic study on simultaneous estimation of picfeltarraenins IA and IB in rats.

  9. Algorithms for the analysis of ensemble neural spiking activity using simultaneous-event multivariate point-process models.

    Science.gov (United States)

    Ba, Demba; Temereanca, Simona; Brown, Emery N

    2014-01-01

    Understanding how ensembles of neurons represent and transmit information in the patterns of their joint spiking activity is a fundamental question in computational neuroscience. At present, analyses of spiking activity from neuronal ensembles are limited because multivariate point process (MPP) models cannot represent simultaneous occurrences of spike events at an arbitrarily small time resolution. Solo recently reported a simultaneous-event multivariate point process (SEMPP) model to correct this key limitation. In this paper, we show how Solo's discrete-time formulation of the SEMPP model can be efficiently fit to ensemble neural spiking activity using a multinomial generalized linear model (mGLM). Unlike existing approximate procedures for fitting the discrete-time SEMPP model, the mGLM is an exact algorithm. The MPP time-rescaling theorem can be used to assess model goodness-of-fit. We also derive a new marked point-process (MkPP) representation of the SEMPP model that leads to new thinning and time-rescaling algorithms for simulating an SEMPP stochastic process. These algorithms are much simpler than multivariate extensions of algorithms for simulating a univariate point process, and could not be arrived at without the MkPP representation. We illustrate the versatility of the SEMPP model by analyzing neural spiking activity from pairs of simultaneously-recorded rat thalamic neurons stimulated by periodic whisker deflections, and by simulating SEMPP data. In the data analysis example, the SEMPP model demonstrates that whisker motion significantly modulates simultaneous spiking activity at the 1 ms time scale and that the stimulus effect is more than one order of magnitude greater for simultaneous activity compared with non-simultaneous activity. Together, the mGLM, the MPP time-rescaling theorem and the MkPP representation of the SEMPP model offer a theoretically sound, practical tool for measuring joint spiking propensity in a neuronal ensemble.

  10. A novel simultaneous measurement method to assess the influence of intracerebroventricular obestatin on colonic motility and secretion in conscious rats.

    Science.gov (United States)

    Chen, Chih-Yen; Doong, Ming-Luen; Li, Chung-Pin; Liaw, Wen-Jinn; Lee, Hsing-Feng; Chang, Full-Young; Lin, Han-Chieh; Lee, Shou-Dong

    2010-06-01

    Obestatin, a novel putative 23-amino acid peptide, is derived from mammalian preproghrelin gene via a bioinformatics approach. Although obestatin regulates thirst, sleep, memory, anxiety, activates cortical neurons in the brain and stimulate proliferation of retinal pigment epithelial cells, there is no study to explore its central impacts on the lower gut motility and secretion. We investigated the influence of intracerebroventricular (ICV) injection of obestatin on rat colonic motor and secretory functions. Colonic transit time, fecal pellet output and fecal content were assessed in freely fed, conscious rats, which were implanted with ICV and colonic catheters chronically. Human/rat corticotropin-releasing factor (h/rCRF) was applied as a stimulatory inducer of colonic motility and secretion. ICV injection of obestatin (0.1, 0.3, 1.0 nmol/rat) did not modify the colonic transit time, whereas ICV injection of h/rCRF (0.3 nmol/rat) significantly shortened colonic transit time. ICV obestatin in any dose we tested did not affect the fecal pellet output, frequency of watery diarrhea, total fecal weight, fecal dried solid weight, or fecal fluid weight in the first hour post-injection, either. In contrast, ICV injection of h/rCRF effectively stimulated fecal pellet output, as well as increased total fecal weight, fecal dried solid weight and fecal fluid weight during the first hour post-injection, compared to ICV saline controls. In conclusion, using our novel simultaneous measurement method, acutely central administration of obestatin exhibits no influence on colonic motility and secretion in conscious rats.

  11. Simultaneous determination of borneol and its metabolite in rat plasma by GC-MS and its application to pharmacokinetic study

    Institute of Scientific and Technical Information of China (English)

    Xiu-Man Sun; Qiong-Feng Liao; Yu-Ting Zhou; Xue-Jiao Deng; Zhi-Yong Xie

    2014-01-01

    A gas chromatography mass spectrometry (GC-MS) method has been developed and fully validated for the simultaneous determination of natural borneol (NB) and its metabolite, camphor, in rat plasma. Following a single liquid-liquid extraction, the analytes were separated using an HP-5MS capillary column (0.25 mm ? 30 m ? 0.25μm) and analyzed by MS in the selected ion monitoring mode. Selected ion monitor (m/z) of borneol, camphor and internal standard was 95, 95 and 128, respectively. Linearity, accuracy, precision and extraction recovery of the analytes were all satisfactory. The method was successfully applied to pharmacokinetic studies of NB after oral administration to Wistar rats.

  12. Simultaneous determination of borneol and its metabolite in rat plasma by GC–MS and its application to pharmacokinetic study

    Directory of Open Access Journals (Sweden)

    Xiu-Man Sun

    2014-10-01

    Full Text Available A gas chromatography mass spectrometry (GC–MS method has been developed and fully validated for the simultaneous determination of natural borneol (NB and its metabolite, camphor, in rat plasma. Following a single liquid–liquid extraction, the analytes were separated using an HP-5MS capillary column (0.25 mm×30 m×0.25 μm and analyzed by MS in the selected ion monitoring mode. Selected ion monitor (m/z of borneol, camphor and internal standard was 95, 95 and 128, respectively. Linearity, accuracy, precision and extraction recovery of the analytes were all satisfactory. The method was successfully applied to pharmacokinetic studies of NB after oral administration to Wistar rats.

  13. An HPLC-MS/MS method for simultaneous determination of decitabine and its valyl prodrug valdecitabine in rat plasma.

    Science.gov (United States)

    Zhang, Youxi; Sun, Jin; Gao, Yikun; Kong, Ying; Xu, Youjun; Jia, Weiru; Liao, Chuanrong; Zhang, Peng; Lian, He; Han, Xiaopeng; Li, Dongpo; Geng, Yajie; He, Zhonggui

    2013-02-15

    A simple and sensitive HPLC-MS/MS method was developed and validated for the simultaneous determination of decitabine and valdecitabine in rat plasma. The analytes were separated on a C(18) column (150mm×4.6mm, 3.5μm) and a triple-quadrupole mass spectrometer equipped with an electrospray ionization (ESI) source was applied for detection. A clean solid-phase extraction procedure with cation exchange cartridge was employed to extract the analytes from rat plasma with high recovery of decitabine (>82%). The calibration curves were linear over a concentration range of 10-10,000ng/mL for decitabine and 5-500ng/mL for valdecitabine. The lower limit of quantitation (LLOQ) of decitabine and valdecitabine was 10 and 5ng/mL, respectively. The intra-day and inter-day precisions were less than 15% and the relative error (RE) was all within ±15%. The validated method was successfully applied to a pharmacokinetics study in rats after either decitabine or valdecitabine orally administrated to the Sprague-Dawley rats.

  14. Quantitative Method for Simultaneous Analysis of Acetaminophen and 6 Metabolites.

    Science.gov (United States)

    Lammers, Laureen A; Achterbergh, Roos; Pistorius, Marcel C M; Romijn, Johannes A; Mathôt, Ron A A

    2017-04-01

    Hepatotoxicity after ingestion of high-dose acetaminophen [N-acetyl-para-aminophenol (APAP)] is caused by the metabolites of the drug. To gain more insight into factors influencing susceptibility to APAP hepatotoxicity, quantification of APAP and metabolites is important. A few methods have been developed to simultaneously quantify APAP and its most important metabolites. However, these methods require a comprehensive sample preparation and long run times. The aim of this study was to develop and validate a simplified, but sensitive method for the simultaneous quantification of acetaminophen, the main metabolites acetaminophen glucuronide and acetaminophen sulfate, and 4 Cytochrome P450-mediated metabolites by using liquid chromatography with mass spectrometric (LC-MS) detection. The method was developed and validated for the human plasma, and it entailed a single method for sample preparation, enabling quick processing of the samples followed by an LC-MS method with a chromatographic run time of 9 minutes. The method was validated for selectivity, linearity, accuracy, imprecision, dilution integrity, recovery, process efficiency, ionization efficiency, and carryover effect. The method showed good selectivity without matrix interferences. For all analytes, the mean process efficiency was >86%, and the mean ionization efficiency was >94%. Furthermore, the accuracy was between 90.3% and 112% for all analytes, and the within- and between-run imprecision were method presented here enables the simultaneous quantification of APAP and 6 of its metabolites. It is less time consuming than previously reported methods because it requires only a single and simple method for the sample preparation followed by an LC-MS method with a short run time. Therefore, this analytical method provides a useful method for both clinical and research purposes.

  15. Simple liquid chromatographic method for the rapid and simultaneous determination of propoxur and its major metabolite isopropoxy phenol in rat blood and urine using solid-phase extraction.

    Science.gov (United States)

    Suma, Ramagiri; Sarin, R K; Saiprakash, P K; Ramakrishna, Sistla

    2005-10-01

    This research paper describes the development and validation of an analytical method for the simultaneous determination of propoxur and isopropoxy phenol (IPP, a major metabolite) in both blood and urine of rat using reversed-phase high-performance liquid chromatography (HPLC) employing solid-phase extraction (SPE). Sample purification was performed using a weak cation-exchange cartridge (Isolute CBA). Separation was achieved by HPLC with UV detection at 270 nm. Recoveries of propoxur and IPP from blood and urine by SPE exceeded 85%. The validated calibration range for propoxur is from 0.5 to 100 microg/L and 2 to 100 microg/L for IPP in both rat blood and urine. The limit of quantitation for propoxur in blood and urine is 0.5 and 0.8 pg/L, respectively, and 2.0 and 4.2 microg/L, respectively, for IPP. Validation results on specificity, sensitivity, linearity, precision, accuracy, and stability are shown. The applicability of the method was demonstrated by the analysis of urine and blood from rats that were orally fed propoxur at minimum dose.

  16. Simultaneous Detection of Group A Rotavirus in Swine and Rat on a Pig Farm in Brazil

    Directory of Open Access Journals (Sweden)

    Paloma de Oliveira Tonietti

    2013-01-01

    Full Text Available This study investigated the occurrence of rotavirus in porcine and Rattus norvegicus, at the same time, on a pig farm in the city of Jaguariúna, São Paulo, Brazil. Swine ( and rat ( fecal samples were analyzed by nested RT-PCR assay. Rotavirus occurred in seven porcine and two rat samples. A total of three pig and one rat samples were further submitted to genetic sequencing. The partial NSP5 gene phylogeny showed that all strains were segregated in the genotype H1. These results point toward a cross-species transmission between rats and pigs on the surveyed farm and represent the first detection of rotavirus in Rattus norvegicus in Brazil.

  17. The effect of NMDA-R antagonism on simultaneously acquired local field potentials and tissue oxygen levels in the brains of freely-moving rats.

    Science.gov (United States)

    Kealy, John; Commins, Sean; Lowry, John P

    2017-01-11

    Non-competitive NMDA receptor antagonists are known to induce psychosis-like symptoms in rodents. Administration of such compounds cause behavioural effects such as memory impairment and hyperlocomotion. Additionally, drugs such as phencyclidine (PCP), ketamine and MK-801 all cause distinctive increases in striatal local field potential (LFP) in the high frequency oscillation (HFO) band in the power spectrum (140-180 Hz). Amperometric sensors provide a means to measure tissue oxygen (tO2; a BOLD-like signal) in the brains of freely-moving rats while simultaneously acquiring LFP using the same electrode. Carbon paste electrodes were implanted into the striatum and hippocampus of male Wistar rats. Rats were administered with saline, ketamine (10 mg/kg), MK-801 (0.1 mg/kg) and PCP (2.5 mg/kg) and recordings were made at 1 kHz using three different potentials (-650 mV to measure tO2; 0 mV and +700 mV as control conditions). NMDA receptor antagonism caused significant increases in tO2 in both the striatum and the hippocampus. Power spectrum analysis showed significant increases in HFO power in the striatum but not in the hippocampus. Conversely, there were significant decreases in delta and alpha power along with increases in theta and gamma power in the hippocampus that were absent in the striatum. This supports findings that LFP can be obtained from an amperometric sensor signal; allowing simultaneous acquisition of two translational biomarkers of neuronal activity (LFP and tO2).

  18. Simultaneous determination of pregnenolone sulphate, dehydroepiandrosterone and allopregnanolone in rat brain areas by liquid chromatography-electrospray tandem mass spectrometry.

    Science.gov (United States)

    Rustichelli, Cecilia; Pinetti, Diego; Lucchi, Chiara; Ravazzini, Federica; Puia, Giulia

    2013-07-01

    Neurosteroids (NSs) are well known modulators of neuronal activity and by binding to different neuronal receptors are responsible for a broad spectrum of biological and pathophysiological conditions. Here, a sensitive liquid chromatographic-electrospray ionization-tandem mass spectrometric method (LC-ESI-MS/MS) has been developed and validated for the simultaneous determination in rat brain areas of three NSs, i.e. pregnenolone sulphate (PS), dehydroepiandrosterone (DHEA) and allopregnanolone (AP). NSs were extracted with methanol-formic acid, purified by Hybrid-SPE cartridges and subjected to LC-ESI-MS/MS without any preliminary derivatization or deconjugation procedure. Quantitation was performed by multiple reaction monitoring mode with the internal standard method, using deuterium-labelled analogues of the analyzed NSs. The proposed method provided for the first time a direct quantitative determination of PS without hydrolysis; in particular, PS concentrations were found significantly (pmemory, than in cortical tissue of control rats, suggesting the important role of this NS in the process of memory formation. The developed method could be successfully applied to quantify simultaneously PS, DHEA and AP levels in brain tissue in order to study their changes during various neurodegenerative diseases and to investigate the role of PS in the brain. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Analgesic effect of simultaneous exposure to infrared laser radiation and μT magnetic field in rats

    Science.gov (United States)

    Cieslar, Grzegorz; Mrowiec, Janina; Kasperczyk, Slawomir; Sieron-Stoltny, Karolina; Sieron, Aleksander

    2008-03-01

    The aim of the experiment was to estimate the effect of repeated simultaneous exposures to infrared laser radiation and μT variable magnetic field used in magnetostimulation on pain perception in rats, as well as the involvement of endogenous opioid system in the mechanism of this effect. In experimental group clean-shaven scull of male Wistar rats placed individually in a specially designed plastic chamber were simultaneously exposed to infrared laser radiation (wavelength - 855 nm, mean power - 4,1 mW, energy density - 30 J/cm2) and variable magnetic field of saw-like shape of impulse, at a frequency of basic impulse 180-195 Hz and mean induction value of 120 μT generated by magneto-laser applicator of device for magnetostimulation Viofor JPS (Med & Life, Poland) 12 minutes daily for 2 periods of 5 consecutive days, with 2 days-lasting break between them, while control animals were sham-exposed. The pain perception was determined by means of "hot plate" test on the basis of calculated analgesic index. As a result of repeated exposures a significant increase in analgesic index persisting also till 14 th day after the end of a cycle of exposures was observed. This analgesic effect was inhibited by prior i.p. injection of opioid antagonist - Naloxone.

  20. Peer influences on drug self-administration: an econometric analysis in socially housed rats.

    Science.gov (United States)

    Peitz, Geoffrey W; Strickland, Justin C; Pitts, Elizabeth G; Foley, Mark; Tonidandel, Scott; Smith, Mark A

    2013-04-01

    Social-learning theories of substance use propose that members of peer groups influence the drug use of other members by selectively modeling, reinforcing, and punishing either abstinence-related or drug-related behaviors. The objective of the present study was to examine the social influences on cocaine self-administration in isolated and socially housed rats, under conditions where the socially housed rats were tested simultaneously with their partner in the same chamber. To this end, male rats were obtained at weaning and housed in isolated or pair-housed conditions for 6 weeks. Rats were then implanted with intravenous catheters and cocaine self-administration was examined in custom-built operant conditioning chambers that allowed two rats to be tested simultaneously. For some socially housed subjects, both rats had simultaneous access to cocaine; for others, only one rat of the pair had access to cocaine. An econometric analysis was applied to the data, and the reinforcing strength of cocaine was measured by examining consumption (i.e. quantity demanded) and elasticity of demand as a function of price, which was manipulated by varying the dose and ratio requirements on a fixed ratio schedule of reinforcement. Cocaine consumption decreased as a function of price in all groups. Elasticity of demand did not vary across groups, but consumption was significantly lower in socially housed rats paired with a rat without access to cocaine. These data suggest that the presence of an abstaining peer decreases the reinforcing strength of cocaine, thus supporting the development of social interventions in drug abuse prevention and treatment programs.

  1. Simultaneous Quantification of Baricitinib and Methotrexate in Rat Plasma by LC-MS/MS: Application to a Pharmacokinetic Study.

    Science.gov (United States)

    Veeraraghavan, Sridhar; Thappali, Satheeshmanikandan R S; Viswanadha, Srikant; Vakkalanka, Swaroop; Rangaswamy, Manivannan

    2016-01-01

    Efficacy assessments using a combination of baricitinib and methotrexate necessitate the development of an analytical method for the determination of both drugs in plasma with precision. A high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of baricitinib and methotrexate in rat plasma. Extraction of baricitinib, methotrexate, and tolbutamide (internal standard; IS) from 50 µL of rat plasma was carried out by protein precipitation with methanol. Chromatographic separation of the analytes was performed on the YMC pack ODS AM (150 mm × 4.6 mm, 5 µm) column under gradient conditions with methanol: 2.0 mM ammonium acetate buffer as the mobile phases at a flow rate of 1 mL/min. The precursor ion and product ion transition for both analytes and IS were monitored on a triple quadrupole mass spectrometer, operated with selective reaction monitoring in positive ionization mode. The method was validated over a concentration range of 0.5-250.00 ng/mL for baricitinib and methotrexate. Mean extraction recoveries for baricitinib, methotrexate, and IS of 86.8%, 89.4%, and 91.8% were consistent across low, medium, and high QC levels, respectively. Precision and accuracy at low, medium, and high quality control levels were less than 15% across the analytes. Benchtop, wet, freeze-thaw, and long-term stability were evaluated for both of the analytes. The analytical method was applied to support the pharmacokinetic study of simultaneous estimation of baricitinib and methotrexate in Wistar rats. Assay reproducibility was demonstrated by reanalysis of 18 incurred samples.

  2. Long-term Statistical Analysis of the Simultaneity of Forbush Decrease Events at Middle Latitudes

    Science.gov (United States)

    Lee, Seongsuk; Oh, Suyeon; Yi, Yu; Evenson, Paul; Jee, Geonhwa; Choi, Hwajin

    2015-03-01

    Forbush Decreases (FD) are transient, sudden reductions of cosmic ray (CR) intensity lasting a few days, to a week. Such events are observed globally using ground neutron monitors (NMs). Most studies of FD events indicate that an FD event is observed simultaneously at NM stations located all over the Earth. However, using statistical analysis, previous researchers verified that while FD events could occur simultaneously, in some cases, FD events could occur non-simultaneously. Previous studies confirmed the statistical reality of non-simultaneous FD events and the mechanism by which they occur, using data from high-latitude and middle-latitude NM stations. In this study, we used long-term data (1971-2006) from middle-latitude NM stations (Irkutsk, Climax, and Jungfraujoch) to enhance statistical reliability. According to the results from this analysis, the variation of cosmic ray intensity during the main phase, is larger (statistically significant) for simultaneous FD events, than for non-simultaneous ones. Moreover, the distribution of main-phase-onset time shows differences that are statistically significant. While the onset times for the simultaneous FDs are distributed evenly over 24- hour intervals (day and night), those of non-simultaneous FDs are mostly distributed over 12-hour intervals, in daytime. Thus, the existence of the two kinds of FD events, according to differences in their statistical properties, were verified based on data from middle-latitude NM stations.

  3. Simultaneous determination of selected eicosanoids by reversed-phase HPLC method using fluorescence detection and application to rat and human plasma, and rat heart and kidney samples.

    Science.gov (United States)

    Aghazadeh-Habashi, Ali; Asghar, Waheed; Jamali, Fakhreddin

    2015-06-10

    Eicosanoids are biologically active lipid-derived oxidative metabolites of arachidonic acid. We, herein, present an improved sensitive, selective and robust high performance liquid chromatography (HPLC)-fluorescence assay for simultaneous quantification of eicosanoids in human plasma and rat tissues. Aliquots of 200 μL of plasma or 30 mg of heart or kidney tissues were spiked with 16-hydroxydecanoic acid as internal standard, and extracted with anhydrous acetonitrile using solid phase cartridges. The eluted samples were dried, reconstituted in anhydrous acetonitrile and labeled with 2-(2,3-naphthalimino)ethyl-trifluoromethanesulphonate in the presence of saturated potassium fluoride solution in anhydrous acetonitrile and N,N-diiospropylethylamine as catalyst. The derivatized eicosanoids were extracted with anhydrous acetonitrile using solid phase cartridges. Chromatographic separation was achieved on a C18 reversed phase column using gradient mobile phase of 0.05% of formic acid:acetonitrile:water at 0.8 mL/min flow rate. The analytes were detected at excitation and emission wavelength of 260 and 396 nm, respectively. The assay was linear (r(2)≥ 0.98) in the concentration range of 0.01-2.5 μg/mL. The intra-day and inter-day coefficients variation was less than 19.8%. Using this assay, we were able to quantify arachidonic acid metabolites simultaneously in human and rat biological samples.

  4. Missing values in multi-level simultaneous component analysis

    NARCIS (Netherlands)

    Josse, Julie; Timmerman, Marieke E.; Kiers, Henk A. L.

    2013-01-01

    Component analysis of data with missing values is often performed with algorithms of iterative imputation. However, this approach is prone to overfitting problems. As an alternative, Josse et al. (2009) proposed a regularized algorithm in the framework of Principal Component Analysis (PCA). Here we

  5. Water Pipeline Network Analysis Using Simultaneous Loop Flow ...

    African Journals Online (AJOL)

    2013-03-01

    Mar 1, 2013 ... A water pipeline network analysis with a case study of Owerri in Imo ... relatively complex problem, particularly if the ...... [11] Mason, P.R., Patterson, B.A. & Loewenson, R. (1986) “Piped water supply and Intestinal parasitism.

  6. Simultaneous analysis and quality assurance for diffusion tensor imaging.

    Directory of Open Access Journals (Sweden)

    Carolyn B Lauzon

    Full Text Available Diffusion tensor imaging (DTI enables non-invasive, cyto-architectural mapping of in vivo tissue microarchitecture through voxel-wise mathematical modeling of multiple magnetic resonance imaging (MRI acquisitions, each differently sensitized to water diffusion. DTI computations are fundamentally estimation processes and are sensitive to noise and artifacts. Despite widespread adoption in the neuroimaging community, maintaining consistent DTI data quality remains challenging given the propensity for patient motion, artifacts associated with fast imaging techniques, and the possibility of hardware changes/failures. Furthermore, the quantity of data acquired per voxel, the non-linear estimation process, and numerous potential use cases complicate traditional visual data inspection approaches. Currently, quality inspection of DTI data has relied on visual inspection and individual processing in DTI analysis software programs (e.g. DTIPrep, DTI-studio. However, recent advances in applied statistical methods have yielded several different metrics to assess noise level, artifact propensity, quality of tensor fit, variance of estimated measures, and bias in estimated measures. To date, these metrics have been largely studied in isolation. Herein, we select complementary metrics for integration into an automatic DTI analysis and quality assurance pipeline. The pipeline completes in 24 hours, stores statistical outputs, and produces a graphical summary quality analysis (QA report. We assess the utility of this streamlined approach for empirical quality assessment on 608 DTI datasets from pediatric neuroimaging studies. The efficiency and accuracy of quality analysis using the proposed pipeline is compared with quality analysis based on visual inspection. The unified pipeline is found to save a statistically significant amount of time (over 70% while improving the consistency of QA between a DTI expert and a pool of research associates. Projection of QA

  7. Simultaneous analysis and quality assurance for diffusion tensor imaging.

    Science.gov (United States)

    Lauzon, Carolyn B; Asman, Andrew J; Esparza, Michael L; Burns, Scott S; Fan, Qiuyun; Gao, Yurui; Anderson, Adam W; Davis, Nicole; Cutting, Laurie E; Landman, Bennett A

    2013-01-01

    Diffusion tensor imaging (DTI) enables non-invasive, cyto-architectural mapping of in vivo tissue microarchitecture through voxel-wise mathematical modeling of multiple magnetic resonance imaging (MRI) acquisitions, each differently sensitized to water diffusion. DTI computations are fundamentally estimation processes and are sensitive to noise and artifacts. Despite widespread adoption in the neuroimaging community, maintaining consistent DTI data quality remains challenging given the propensity for patient motion, artifacts associated with fast imaging techniques, and the possibility of hardware changes/failures. Furthermore, the quantity of data acquired per voxel, the non-linear estimation process, and numerous potential use cases complicate traditional visual data inspection approaches. Currently, quality inspection of DTI data has relied on visual inspection and individual processing in DTI analysis software programs (e.g. DTIPrep, DTI-studio). However, recent advances in applied statistical methods have yielded several different metrics to assess noise level, artifact propensity, quality of tensor fit, variance of estimated measures, and bias in estimated measures. To date, these metrics have been largely studied in isolation. Herein, we select complementary metrics for integration into an automatic DTI analysis and quality assurance pipeline. The pipeline completes in 24 hours, stores statistical outputs, and produces a graphical summary quality analysis (QA) report. We assess the utility of this streamlined approach for empirical quality assessment on 608 DTI datasets from pediatric neuroimaging studies. The efficiency and accuracy of quality analysis using the proposed pipeline is compared with quality analysis based on visual inspection. The unified pipeline is found to save a statistically significant amount of time (over 70%) while improving the consistency of QA between a DTI expert and a pool of research associates. Projection of QA metrics to a low

  8. Analysis of Heme oxygenase isomers in rat

    Institute of Scientific and Technical Information of China (English)

    Zhen-Wei Xia; Wen-Jun Cui; Xue-Hong Zhang; Qing-Xiang Shen; Jian Wang; Yun-Zhu Li; Shen-Nian Chen; Shan-Chang Yu

    2002-01-01

    AIM: To purify and identify heme oxygenase (HO) isomers which exist in rat liver, spleen and brain treated with hematin and phenylhydrazine and in untreated rat liver and to investigate the characteristics of HO isomers, to isolate and confirm the rat HO-1 cDNA that actually encodes HO-1 by expressing cDNA in monkey kidney cells (COS-1 cells), to prepare the rat heme oxygenase-1 (HO-1) mutant and to detect inhibition of HO-1 mutated enzyme.METHODS: First, rat liver, spleen and brain microsomal fi-actions were purified by DEAE-Sephacel and hydroxylapatite. The characteristics including activity, immunity and inducibility of two isomers (HO-1 and HO-2), and their apparent molecular weight were measured by detecting enzymatic activities, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis, respectively. Second, plasmid pcDNA3HO1 containing native rat HO-1 cDNA and pcDNA3HO1D25 carrying mutated rat HO-1 cDNA (His25Ala) were constructed by site-directed mutagenesis. COS-1 cells transfected with pcDNA3HO1 and pcDNA3HO1D25 were collected and disrupted by sonication, the microsomes were prepared by ultracentrifugation. Third, the inhibition of rat HO-1 mutant was analyzed.RESULTS: Two isomers were purified and identified in treated rat liver, spleen, brain and untreated rat liver. HO-1 was the predominant form with a ratio of 2.0:1 and 3.2:1 of HO-1 and HO-2 in liver and spleen, respectively, but only the activity of HO-2 in the brain and untreated liver could be detected. The apparent molecular weights of HO-1 and HO-2 were about Mr 30 000 and Mr 36 000 under reducing conditions, respectively. The antiserum against liver HO-2 was employed in Western blotting analysis, the reactivity of HO-1 in the liver was not observed. The plasmid pcDNA3HO1 was highly expressed in endoplasmic reticulum of transfected COS-1 cells. The specific activity was ≈5-fold higher than that of the control. However, the enzyme activity of mutated HO-1 declined. While

  9. Simultaneous quantification of two steroidal glycosides after oral gavage of Marsdenia tenacissima extract in rats using a LC-MS/MS method.

    Science.gov (United States)

    Wang, Wei; Sun, Wei; Dong, Lei; Qu, Xiaotong; Shi, Jin; Li, Ke-Jun; Li, Bing; Ma, Da-Peng

    2015-04-01

    A specific, sensitive and accurate analytical LC-MS/MS assay was developed for the simultaneous determination of two steroidal glycosides, tenacissoside H and tenacissoside I, in rat plasma. An Agilent ZORBAX SB-C18 column was used with an isocratic mobile phase system composed of methanol-water-formic acid (70:30:0.1, v/v/v) at a flow rate of 0.3 mL/min. The analysis was performed on a positive ionization electrospray mass spectrometer via selected reaction monitoring mode scan. One-step protein precipitation with acetonitrile was chosen to extract the analytes from plasma. The lower limits of quantification were 0.9 ng/mL for tenacissoside H and tenacissoside I. The intra- and inter-day precisions were 2.03-11.56 and 3.76-11.62%, respectively, and the accuracies were method was applied to a pharmacokinetic study in rats after oral gavage of Marsdenia tenacissima extract.

  10. High-throughput liquid chromatography tandem mass spectrometry method for simultaneous determination of fampridine, paroxetine, and quinidine in rat plasma: Application to in vivo perfusion study

    Directory of Open Access Journals (Sweden)

    Suneetha Achanti

    2016-10-01

    Full Text Available A selective and high-throughput liquid chromatography–mass spectrometry method has been developed and validated for the simultaneous quantification of paroxetine, fampridine, and quinidine in rat plasma using imipramine as an internal standard. Following protein precipitation extraction, the analytes and internal standard were run on XBridge C18 column (150 mm × 4.6 mm, 5 μm using a gradient mobile phase consisting of 5mM ammonium formate in water (pH 9.0 and acetonitrile in a flow gradience program. The precursor and product ions of the drugs were monitored on a triple quadrupole instrument operated in the positive ionization mode. The method was validated over a concentration range of 0.1–100 ng/mL for all the three analytes, with relative recoveries ranging from 69% to 82%. The intra- and interbatch precision (percent coefficient of variation across four validation runs were less than 13.4%. The accuracy determined at four quality control (QC levels (lower limit of quantitation, low QC, medium QC, and high QC was within ±6.5% of coefficient of variation values. The method proved highly reproducible and sensitive, and was successfully applied in a pharmacokinetic study after single-dose oral administration to rats and also in perfusion study sample analysis.

  11. Simultaneous transplantation of fetal ventral mesencephalic tissue and encapsulated genetically modified cells releasing GDNF in a hemi-parkinsonian rat model of Parkinson's disease

    DEFF Research Database (Denmark)

    Perez-Bouza, Alberto; Di Santo, Stefano; Seiler, Stefanie

    2017-01-01

    Transplantation of fetal ventral mesencephalic (VM) neurons for Parkinson's disease (PD) is limited by poor survival and suboptimal integration of grafted tissue into the host brain. In a 6-OHDA rat model of PD we investigated the feasibility of simultaneous transplantation of rat fetal VM tissue...... and polymer-encapsulated C2C12 myoblasts genetically modified to produce glial cell line-derived neurotrophic factor or mocktransfected myoblasts on graft function. Amphetamine-induced rotations were assessed prior and 2, 4, 6 and 9 weeks post-transplantation. We found that rats grafted with VM transplants...

  12. Simultaneous quantification of caffeine and its three primary metabolites in rat plasma by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Choi, Eu Jin; Bae, Soo Hyeon; Park, Jung Bae; Kwon, Min Jo; Jang, Su Min; Zheng, Yu Fen; Lee, Young Sun; Lee, Su-Jun; Bae, Soo Kyung

    2013-12-01

    A rapid, sensitive, simple and accurate LC-MS/MS method for the simultaneous quantitation of caffeine, and its three primary metabolites, theobromine, paraxanthine, and theophylline, in rat plasma was developed and validated. Chromatographic separation was performed on an Agilent Poroshell 120 EC-C18 column using 1 μg/mL acetaminophen as an internal standard. Each sample was run at 0.5 mL/min for a total run time of 7 min/sample. Detection and quantification were performed using a mass spectrometer in selected reaction-monitoring mode with positive electrospray ionization. The lower limit of quantification was 5 ng/mL for all analytes with linear ranges up to 5000 ng/mL for caffeine and 1000 ng/mL for its metabolites. The coefficient of variation for assay precision was less than 12.6%, with an accuracy of 93.5-114%. The assay was successfully applied to determine plasma concentrations of caffeine, theobromine, paraxanthine, and theophylline in rat administered various energy drinks containing the same caffeine content. Various energy drinks exhibited considerable variability in the pharmacokinetic profiles of caffeine and its three primary metabolites, even containing the same caffeine. Different additives of energy drinks might contribute to these results. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Simultaneous determination of N(1)-acetyl sulfisoxazole and its metabolites, and relative bioavailability compare to sulfisoxazole in rats.

    Science.gov (United States)

    Kim, Eunyoung; Kang, Wonku

    2016-09-10

    N(1)-acetyl sulfisoxazole (N1AS), a dihydropteroate synthase inhibitor is known to be biotransformed primarily to sulfisoxazole, partly to N(4)-acetyl sulfisoxazole (N4AS), and likely also to diacetyl sulfisoxazole (DAS) and other compounds. Although its clinical use has been limited due to urolithiasis, some countries still use the drug in combination with trimethoprim in cattle. A liquid chromatographic method using ultraviolet detection was developed for the simultaneous determination of four substances for the first time. Four analytes and sulfamethoxazole (IS) were separated on a reversed-phase column with gradient elution of a mobile phase. Because DAS and N1AS in plasma were changed very quickly into N4AS and sulfisoxazole, respectively, and esterase inhibitors (sodium fluoride and eserine) could not prevent the transformation, sulfisoxazole and N4AS were monitored in rat plasma following a single oral administration of N1AS and sulfisoxazole in five rats. The relative bioavailability of N1AS to sulfisoxazole was about two, indicating that a half-dose of N1AS would be equivalent to a dose of sulfisoxazole to achieve the same systemic exposure to sulfisoxazole.

  14. A novel detachable head-mounted device for simultaneous EEG and photoacoustic monitoring of epilepsy in freely moving rats.

    Science.gov (United States)

    Wang, Bo; Zhou, Junli; Carney, Paul; Jiang, Huabei

    2015-02-01

    The study of neuro-hemodynamic changes in freely moving animals provides for a better understanding of brain dynamics in normal and disease states. While it has been shown that hemodynamic changes are closely related to seizures, methods for detection in freely moving animals are limited. In this work, we integrate photoacoustic sensor technology and electroencephalography into a small portable device that can be attached on the head of wake freely moving animals. We demonstrate chronic simultaneous monitoring of photoacoustic and electroencephalographic signals in an acute seizure model of epilepsy. Our results demonstrate that both the neural and vascular responses during seizures in freely moving rats have characteristics which are observed to be different and more diverse from that of anesthetized rats. This implies that the neurovascular coupling in seizure in free moving animals are more complicated, which calls for more detailed study in future. To the best of our knowledge, this is the first time for hemodynamic monitoring of seizure in free moving animals. This technology also promises for other hemodynamic related research study in freely moving small animals.

  15. Development and validation of UPLC-MS/MS method for simultaneous determination of gestodene and ethinyl estradiol in rat plasma.

    Science.gov (United States)

    Xiong, Zhili; Sun, Xiaohong; Huo, Taoguang; Li, Ning; Zheng, Yan; Sun, Yi

    2010-02-01

    A selective and sensitive ultra-performance liquid chromatography method with tandem mass spectrometric detection for simultaneous determination of gestodene (GES) and ethinyl estradiol (EE) in rat plasma was developed and validated. GES, EE and the internal standard, norgestrel, were extracted with ethyl acetate, derivatized (EE only) with dansyl chloride and then back-extracted into diethyl ether-hexane (2:1, v/v). The separation was performed on an ACQUITY UPLC BEH C(18) column with gradient elution using mobile phase consisting of acetonitrile and water (both containing 0.1% formic acid). The detection was carried out by means of electrospray ionization (ESI) mass spectrometry in positive ion mode with multiple-reaction monitoring. Calibration curves of GES and EE were linear (r(2) >or= 0.99) over the concentration ranges 1.59-159 and 0.196-78.4 ng/mL, respectively. The intra- and inter-day precisions were not more than 6.9 and 12.9% for GES and 10.6 and 9.0% for EE, and the accuracies were -2.5-8.0% for GES, and -7.2-0.19% for EE, respectively. The method herein described was superior to previous methods and was applicable to the pharmacokinetic study of GES and EE in rats.

  16. Simultaneous determination of creatine phosphate, creatine and 12 nucleotides in rat heart by LC-MS/MS.

    Science.gov (United States)

    Wang, Jun-mei; Chu, Yang; Li, Wei; Wang, Xiang-yang; Guo, Jia-hua; Yan, Lu-lu; Ma, Xiao-hui; Ma, Ying-li; Yin, Qi-hui; Liu, Chang-xiao

    2014-05-01

    A simple, rapid and sensitive LC-MS/MS method was developed and validated for simultaneous determination of creatine phosphate (CP), creatine (Cr) and 12 nucleotides in rat heart. The analytes, ATP, ADP, AMP, GTP, GDP, GMP, CTP, CDP, CMP, UTP, UDP, UMP, CP, Cr, were extracted from heart tissue with pre-cooled (0°C) methanol/water (1:1, v/v) and separated on a Hypersil Gold AQ C18 column (150mm×4.6mm, 3μm) using an isocratic elution with a mobile phase consisting of 2mmol/L ammonium acetate in water (pH 10.0, adjusted with ammonia). The detection was performed by negative ion electrospray ionization in selective reaction monitoring mode (SRM). In the assay, all the analytes showed good linearity over the investigated concentration range (r>0.99). The accuracy was between 80.7% and 120.6% and the precision expressed in RSD was less than 15.6%. This method was successfully applied to measure the concentrations of the 12 nucleotides, creatine phosphate and creatine in rat heart for the first time.

  17. In vivo comparison of norepinephrine and dopamine release in rat brain by simultaneous measurements with fast-scan cyclic voltammetry.

    Science.gov (United States)

    Park, Jinwoo; Takmakov, Pavel; Wightman, R Mark

    2011-12-01

    Brain norepinephrine and dopamine regulate a variety of critical behaviors such as stress, learning, memory, and drug addiction. In this study, we demonstrate differences in the regulation of in vivo neurotransmission for dopamine in the anterior nucleus accumbens (NAc) and norepinephrine in the ventral bed nucleus of the stria terminalis (vBNST) of the anesthetized rat. Release of the two catecholamines was measured simultaneously using fast-scan cyclic voltammetry at two different carbon-fiber microelectrodes, each implanted in the brain region of interest. Simultaneous dopamine and norepinephrine release was evoked by electrical stimulation of a region where the ventral noradrenergic bundle, the pathway of noradrenergic neurons, courses through the ventral tegmental area/substantia nigra, the origin of dopaminergic cell bodies. The release and uptake of norepinephrine in the vBNST were both significantly slower than for dopamine in the NAc. Pharmacological manipulations in the same animal demonstrated that the two catecholamines are differently regulated. The combination of a dopamine autoreceptor antagonist and amphetamine significantly increased basal extracellular dopamine whereas a norepinephrine autoreceptor antagonist and amphetamine did not change basal norepinephrine concentration. α-Methyl-p-tyrosine, a tyrosine hydroxylase inhibitor, decreased electrically evoked dopamine release faster than norepinephrine. The dual-microelectrode fast-scan cyclic voltammetry technique along with anatomical and pharmacological evidence confirms that dopamine in the NAc and norepinephrine in the vBNST can be monitored selectively and simultaneously in the same animal. The high temporal and spatial resolution of the technique enabled us to examine differences in the dynamics of extracellular norepinephrine and dopamine concurrently in two different limbic structures.

  18. A computerized system for the simultaneous monitoring of place conditioning and locomotor activity in rats.

    Science.gov (United States)

    Brockwell, N T; Ferguson, D S; Beninger, R J

    1996-02-01

    Place conditioning is one of the most popular behavioral methods for assessing the rewarding properties of various substances. Many substances that are rewarding also influence motor activity. This report describes a computerized system designed to simultaneously monitor both place conditioning and locomotor activity. The system consists of 4 independent conditioning boxes, each equipped with 6 pairs of photosensors connected to an Experiment Controller, an electronic board containing a microprocessor, a programable timer, and 16 K of RAM used to store both instructions and data. The effects of the stimulant (+)-amphetamine were assessed using this system and found to produce a place preference comparable to that obtained from a previously utilized mechanical timer system. The computerized system also demonstrated that amphetamine increased unconditioned activity. There are a number of advantages and broader applications of the new methodology.

  19. Simultaneous telemetric monitoring of brain glucose and lactate and motion in freely moving rats.

    Science.gov (United States)

    Rocchitta, Gaia; Secchi, Ottavio; Alvau, Maria Domenica; Farina, Donatella; Bazzu, Gianfranco; Calia, Giammario; Migheli, Rossana; Desole, Maria Speranza; O'Neill, Robert D; Serra, Pier A

    2013-11-05

    A new telemetry system for simultaneous detection of extracellular brain glucose and lactate and motion is presented. The device consists of dual-channel, single-supply miniature potentiostat-I/V converter, a microcontroller unit, a signal transmitter, and a miniaturized microvibration sensor. Although based on simple and inexpensive components, the biotelemetry device has been used for accurate transduction of the anodic oxidation currents generated on the surface of implanted glucose and lactate biosensors and animal microvibrations. The device was characterized and validated in vitro before in vivo experiments. The biosensors were implanted in the striatum of freely moving animals and the biotelemetric device was fixed to the animal's head. Physiological and pharmacological stimulations were given in order to induce striatal neural activation and to modify the motor behavior in awake, untethered animals.

  20. Microarray Analysis of the Developing Rat Mandible

    Institute of Scientific and Technical Information of China (English)

    Hideo KABURAGI; Naoyuki SUGANO; Maiko OSHIKAWA; Ryosuke KOSHI; Naoki SENDA; Kazuhiro KAWAMOTO; Koichi ITO

    2007-01-01

    To analyze the molecular events that occur in the developing mandible, we examined the expression of 8803 genes from samples taken at different time points during rat postnatal mandible development.Total RNA was extracted from the mandibles of 1-day-old, 1-week-old, and 2-week-old rats. Complementary RNA (cRNA) was synthesized from cDNA and biotinylated. Fragmented cRNA was hybridized to RGU34A GeneChip arrays. Among the 8803 genes tested, 4344 were detectable. We identified 148 genes with significantly increased expression, and 19 genes with significantly decreased expression. A comprehensive analysis appears to be an effective method of studying the complex process of development.

  1. Ca analysis: an Excel based program for the analysis of intracellular calcium transients including multiple, simultaneous regression analysis.

    Science.gov (United States)

    Greensmith, David J

    2014-01-01

    Here I present an Excel based program for the analysis of intracellular Ca transients recorded using fluorescent indicators. The program can perform all the necessary steps which convert recorded raw voltage changes into meaningful physiological information. The program performs two fundamental processes. (1) It can prepare the raw signal by several methods. (2) It can then be used to analyze the prepared data to provide information such as absolute intracellular Ca levels. Also, the rates of change of Ca can be measured using multiple, simultaneous regression analysis. I demonstrate that this program performs equally well as commercially available software, but has numerous advantages, namely creating a simplified, self-contained analysis workflow.

  2. Simultaneous analysis of strychnine and brucine and their major metabolites by liquid chromatography-electrospray ion trap mass spectrometry.

    Science.gov (United States)

    Chen, Xueguo; Lai, Yongquan; Cai, Zongwei

    2012-04-01

    A liquid chromatography-electrospray ionization-ion trap mass spectrometry (LC-ESI-ITMS) method was developed for the simultaneous analysis of strychnine, brucine and their major metabolites. Strychnine and brucine were individually incubated with rat liver S9 fraction. The incubation samples were pooled together and analyzed with LC-ESI-ITMS in positive ion and full-scan detection mode. The calibration curves of strychnine and brucine in rat liver showed good linearity in ranges of 0.020 to 8.0 µg/mL for strychnine and 0.020 to 8.5 µg/mL for brucine. The limits of detections were both 0.008 µg/mL and the recoveries were 88.3 and 83.2% for strychnine and brucine, respectively. Two metabolites were identified as strychnine N-oxide and brucine N-oxide by comparing the molecular mass, retention time, full-scan mass spectra, tandem MS and MS(3) spectra with those of strychnine and brucine. The developed method provided high sensitivity and selectivity for the determination of poisonous alkaloids and their major metabolites and can be applied in the determination of samples in forensic and clinically toxicological cases.

  3. Simultaneous determination of harpagoside and cinnamic acid in rat plasma by liquid chromatography electrospray ionization mass spectrometry and its application to pharmacokinetic studies.

    Science.gov (United States)

    Wang, Su-Jun; Ruan, Jin-Xiu; Zhao, Yan-Hong; Zhang, Zhen-Qing

    2008-01-01

    A simple and sensitive method was developed for the simultaneous quantification of harpagoside and cinnamic acid in rat plasma using high-performance liquid chromatography system coupled to a negative ion electrospray mass spectrometric analysis. The plasma sample preparation was a simple deproteinization by the addition of two volumes of acetonitrile. The analytes were separated on an Intersil C8-3 column (2.1 mm i.d.x250 mm, 5 microm) with acetonitrile-5 mm ammonium formate aqueous solution (60:40, v/v) as mobile phase at a flow-rate of 0.2 mL/min. Detection was performed on a quadrupole mass spectrometer equipped with electrospray ionization (ESI) source operated under selected ion monitoring (SIM) mode. [M+HCOO]- at m/z 539 for harpagoside, [M-H]- at m/z 147 for cinnamic acid and [M-H]- at m/z 137 for salylic acid (internal standard) were selected as detecting ions, respectively. The method was validated over the concentration range 7-250 ng/mL for harpagoside and 5-500 ng/mL for cinnamic acid. The lower limits of quantitation for harpagoside and cinnamic acid were 7 and 5 ng/mL, respectively. The intra- and inter-day precisions (RSD%) were within 9.5% and the assay accuracies (RE%) ranged from -5.3 to 3.0% for both analytes. Their average recoveries were greater than 86%. Both analytes were proved to be stable during all sample storage, preparation and analysis procedures. The method was successfully applied to the pharmacokinetic study of harpagoside and cinnamic acid following oral administration of Radix Scrophulariae extract to rats.

  4. Simultaneous determination of D-aspartic acid and D-glutamic acid in rat tissues and physiological fluids using a multi-loop two-dimensional HPLC procedure.

    Science.gov (United States)

    Han, Hai; Miyoshi, Yurika; Ueno, Kyoko; Okamura, Chieko; Tojo, Yosuke; Mita, Masashi; Lindner, Wolfgang; Zaitsu, Kiyoshi; Hamase, Kenji

    2011-11-01

    For a metabolomics study focusing on the analysis of aspartic and glutamic acid enantiomers, a fully automated two-dimensional HPLC system employing a microbore-ODS column and a narrowbore-enantioselective column was developed. By using this system, a detailed distribution of D-Asp and D-Glu besides L-Asp and L-Glu in mammals was elucidated. For the total analysis concept, the amino acids were first pre-column derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) to be sensitively and fluorometrically detected. For the non-stereoselective separation of the analytes in the first dimension a monolithic ODS column (750 mm × 0.53 mm i.d.) was adopted, and a self-packed narrowbore-Pirkle type enantioselective column (Sumichiral OA-2500S, 250 mm × 1.5 mm i.d.) was selected for the second dimension. In the rat plasma, RSD values for intra-day and inter-day precision were less than 6.8%, and the accuracy ranged between 96.1% and 105.8%. The values of LOQ of D-Asp and D-Glu were 5 fmol/injection (0.625 nmol/g tissue). The present method was successfully applied to the simultaneous determination of free aspartic acid and glutamic acid enantiomers in 7 brain areas, 11 peripheral tissues, plasma and urine of Wistar rats. Biologically significant D-Asp values were found in various tissue samples whereas for D-Glu the values were very low possibly indicating less significance.

  5. Simultaneous determination of gastrodin and puerarin in rat plasma by HPLC and the application to their interaction on pharmacokinetics.

    Science.gov (United States)

    Jiang, Li; Dai, Jundong; Huang, Zhenlin; Du, Qinghua; Lin, Jiahao; Wang, Yurong

    2013-02-01

    Gastrodin (Gas) and puerarin (Pur) are bioactive substances derived from traditional Chinese medicine Gastrodia elata and Radix Puerariae, respectively, which were often used together in Chinese clinical prescriptions. Their injections were used in combined way for treatment of some cardiocerebrovascular diseases in clinic, especially for vertigo due to vertebrobasilar ischemia. In this paper, interaction of gastrodin and puerarin in rat plasma pharmacokinetics via intragastic (i.g.)/intravenous (i.v.) administration was investigated. A reliable HPLC method was developed for simultaneous determination of Gas and Pur in rat plasma with a linear range of 0.101-101 μg/mL for Gas and 0.0500-5.98 μg/mL for Pur (r(2)>0.993). The LLOQ, LOD of Gas and Pur were determined to be 0.101, 0.0486 μg/mL, and 0.05, 0.0245 μg/mL, respectively. The intra-day and inter-day precision were all less than 12.0%, whilst the accuracy were all within 96.4±6.00%. The proposed method has been successfully applied to the pharmacokinetic study of the analytes in rats after i.g./i.v. administration of Gas and Pur alone or combined with each other (i.g.: 40 mg/kg Gas, 400 mg/kg Pur; i.v.: 20 mg/kg Gas, 20 mg/kg Pur). Blood samples were collected from retinal vein plexus of rats at predetermined time points and plasma containing the internal standard tyrosol (IS) were precipitated by methanol and chromatography was carried out on a C(18) column with a gradient mobile phase of ACN-H(2)O with 0.05% phosphoric acid as a modifier. The pharmacokinetic profiles of combined administration were found to be distinct from those of given alone. The C(max), T(max), T(1/2), MRT of Gas administrated alone or combined with Pur via i.g. were 21.7 μg/mL, 0.250 h, 2.81 h, 0.830 h and 18.4 μg/mL, 0.550 h, 0.970 h, 1.37 h, respectively, of Pur administrated alone or combined with Gas via i.g. were 0.490 μg/mL, 1.95 h, 1.33 h, 2.10 h and 2.01 μg/mL, 0.570 h, 4.00 h, 5.10 h, respectively. The relative oral

  6. Simultaneous quantitative analysis of metabolites using ion-pair liquid chromatography-electrospray ionization mass spectrometry

    NARCIS (Netherlands)

    Coulier, L.; Bas, R.; Jespersen, S.; Verheij, E.; Werf, M.J. van der; Hankemeier, T.

    2006-01-01

    We have developed an analytical method, consisting of ion-pair liquid chromatography coupled to electrospray ionization mass spectrometry (IP-LC-ESI-MS), for the simultaneous quantitative analysis of several key classes of polar metabolites, like nucleotides, coenzyme A esters, sugar nucleotides, an

  7. Simultaneous Determination of Arsenic, Manganese, and Selenium in Biological Materials by Neutron-Activation Analysis

    DEFF Research Database (Denmark)

    Heydorn, Kaj; Damsgaard, Else

    1973-01-01

    A new method was developed for the simultaneous determination of arsenic, manganese, and selenium in biological material by thermal-neutron activation analysis. The use of 81 mSe as indicator for selenium permitted a reduction of activation time to 1 hr for a 1 g sample, and the possibility of loss...

  8. Simultaneous Determination and Pharmacokinetics of Metolazone, Losartan and Losartan Carboxylic Acid in Rat Plasma by HPLC-ESI-MS-MS.

    Science.gov (United States)

    Dubey, Ramkumar; Ghosh, Manik; Sinha, Barij Nayan; Muthukrishnan, Venkateswari

    2015-10-01

    For the first time, we developed and validated a highly sensitive, selective and rapid HPLC-ESI-MS-MS method for simultaneous quantification of metolazone (MET), losartan (LOS) and its metabolite losartan carboxylic acid (LCA) in rat plasma. After solid-phase extraction, the analytes and internal standard (irbesartan) were extracted from 100 µL plasma sample on an Agilent Poroshell 120, EC-C18 (50 × 4.6 mm, i.d., 2.7 µm) column using 5 µL injection volume with a total run time of 3 min. Acidified methanol/water mixture was used as a mobile phase. The parent → product ion transitions for MET (m/z 366.0 → 258.9), LOS (m/z 423.2 → 207.0), LCA (m/z 437.0 → 235.1) and IS (m/z 429.2 → 207.0) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring and positive ion mode. The method was found to be linear in the range of 0.05-250 for MET, 2-3,000 for LOS and 4-3,500 ng/mL for LCA. The method was validated with respect to selectivity, linearity, accuracy, precision, recovery and stability according to accepted regulatory guidelines. The described method was successfully applied to preclinical pharmacokinetic studies of analytes after an oral administration of mixture of MET (1 mg/kg) and LOS (10 mg/kg) in rats.

  9. Simultaneous determination of imperatorin and its metabolite xanthotoxol in rat plasma and urine by LC-MS/MS and its application to pharmacokinetic studies.

    Science.gov (United States)

    Ngo, Lien; Tran, Phuong; Ham, Seong-Ho; Cho, Jung-Hee; Cho, Hea-Young; Lee, Yong-Bok

    2017-02-15

    An accurate, precise, selective, and sensitive liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of imperatorin (IMP) and its metabolite, xanthotoxol (XAN), in rat plasma and urine samples. The analytes, along with psoralen as an internal standard, were determined by multiple reaction monitoring (MRM) operated in the positive electrospray ionization (ESI) mode. Chromatographic separation was performed on an Acquity UPLC BEH C18 column (50mm×2.1mm, 1.7μm) with a mobile phase consisting of 0.1% formic acid solution and 0.1% formic acid in methanol at a flow rate of 0.3mL/min. The run time was 6min per sample and the injection volume was 5μL. The method had a lower limit of quantification (LLOQ) of 0.25ng/mL for IMP in plasma and urine, and 1ng/mL for XAN in urine. The linear calibration curves were fitted over the range of 0.25-1000ng/mL for IMP in plasma, 0.25-1000ng/mL for IMP in urine, and 1-1000ng/mL for XAN in urine, with correlation coefficients greater than 0.995. The inter- and intra-day accuracies (relative error, RE%) were between -8.5% and 3.5%, and the precisions (relative standard deviation, RSD%) were less than 10.0% for all quality control samples (QCs). The analytes were extracted from rat plasma and urine samples using a liquid-liquid extraction method with the extraction recovery in the range of 60.3-79.1%. A good stability of the analytes was observed in all the analysis procedures. The method was successfully validated and applied to determine the pharmacokinetics of IMP in rat plasma and, for the first time, the metabolite kinetics of IMP to XAN in rat urine after IMP administration.

  10. Simultaneous activation of gamma and theta network oscillations in rat hippocampal slice cultures.

    Science.gov (United States)

    Fischer, Yacov; Wittner, Lucia; Freund, Tamas F; Gähwiler, Beat H

    2002-03-15

    Hippocampal activity in vivo is characterized by concurrent oscillations at theta (4-15 Hz) and gamma (20-80 Hz) frequencies. Here we show that cholinergic receptor activation (methacholine 10-20 nm) in hippocampal slice cultures induces an oscillatory mode of activity, in which the intrinsic network oscillator (located in the CA3 area) expresses simultaneous theta and gamma network oscillations. Pyramidal cells display synaptic theta oscillations, characterized by cycles consisting of population EPSP-IPSP sequences that are dominated by population IPSPs. These rhythmic IPSPs most probably result from theta-modulated spiking activity of several interneurons. At the same time, the majority of interneurons consistently display synaptic gamma oscillations. These oscillatory cycles consist of fast depolarizing rhythmic events that are likely to reflect excitatory input from CA3 pyramidal cells. Interneurons comprising this functional group were identified morphologically. They include four known types of interneurons (basket, O-LM, bistratified and str. lucidum-specific cells) and one new type of CA3 interneuron (multi-subfield cell). The oscillatory activity of these interneurons is only weakly correlated between neighbouring cells, and in about half of these (44 %) is modulated by depolarizing theta rhythmicity. The overall characteristics of acetylcholine-induced oscillations in slice cultures closely resemble the rhythmicity observed in hippocampal field and single cell recordings in vivo. Both rhythmicities depend on intrinsic synaptic interactions, and are expressed by different cell types. The fact that these oscillations persist in a network lacking extra-hippocampal connections emphasizes the importance of intrinsic mechanisms in determining this form of hippocampal activity.

  11. Performance Analysis of Temperature and Strain Simultaneous Measurement System Based on Heterodyne Detection of Brillouin Scattering

    Institute of Scientific and Technical Information of China (English)

    Ji-Sheng Zhang; Yong-Qian Li; Shuo Zhang; Li-Juan Zhao

    2008-01-01

    Microwave heterodyne detection can be used to measure the temperature and strain distribution along a fiber with high accuracy in a Brillouin optical time domain reflectometry (BOTDR) system. This method involves simultaneous measurement of Brillouin scattering and Rayleigh scattering in fiber, and scanning of Briliouin spectrum to obtain the desired information. This paper presents a simultaneous measurement system of temperature and strain based on microwave detection and analyzed the system performances such as measurement accuracy, dynamic range, and spatial resolution theoretically. The analysis shows that the system can achieve a temperature resolution of 1℃ and a strain resolution of 100 με.

  12. Integrated analysis of transcriptomics and metabonomics profiles in aflatoxin B1-induced hepatotoxicity in rat.

    Science.gov (United States)

    Lu, Xiaoyan; Hu, Bin; Shao, Li; Tian, Yu; Jin, Tingting; Jin, Yachao; Ji, Shen; Fan, Xiaohui

    2013-05-01

    The aim of this work was to identify mechanisms and potential biomarkers for predicting the development and progression of aflatoxin B1 (AFB1)-induced acute hepatotoxicity. In this study, microarray analysis and metabolites profiles were used to identify shifts in gene expression and metabolite levels associated with the affected physiological processes of rats treated with AFB1. Histopathological examinations and serum biochemical analysis were simultaneously performed; the results indicated that hepatotoxicity occurred in higher dosage groups. However, gene expression analysis and metabolite profiles are more sensitive than general toxicity studies for detecting AFB1-induced acute hepatotoxicity as the patterns of low-dose AFB1-treated rats in these two technique platforms were more similar to the rats in higher dosage groups than to the control rats. Integrated analysis of the results from general toxicity studies, transcriptomics and metabonomics profiles suggested that p53 signaling pathway induced by oxidative damage was the crucial step in AFB1-induced acute hepatotoxicity, whereas gluconeogenesis and lipid metabolism disorder were found to be the major metabolic effects after acute AFB1 exposure. The genes and metabolites significantly affected in common in rat liver or serum of three doses AFB1 treatments served as potential biomarkers for detecting AFB1-induced acute hepatotoxicity.

  13. Effectiveness of enterosgel usage in the conditions of simultaneous destruction of rats by karbofos and carbon tetrachloride

    Directory of Open Access Journals (Sweden)

    L. А. Boyko

    2014-10-01

    Full Text Available The aim of our study was to examine the effectiveness of the enterosgel sorbent in the terms of simultaneous destruction of rats by karbofos and carbon tetrachloride. Experiments were carried out on white rats weighing 175–200 g, which were kept on a standard diet. Karbofos was administered intragastrically on daily basis in the aqueous solution at the rate of 20 mg/kg of the body weight, which is 1/10 of the LD50. Carbon tetrachloride was administered intraperitoneally, twice a day in the form of a 50% oil solution at a dose of 1.0 ml/kg of animal. Animals received enterosgel daily by intragastric way at the rate of 120 mg/kg of body weight. The activity of free radical processes in rats was assessed by thiobarbituric acid content – active products (TBA-AP in serum, liver and heart homogenates. Level of endogenous intoxication was determined by the content of average weight molecules (AWM in serum and liver homogenate. The degree of cytolysis of hepatocytes was assessed by the activity of alanine and aspartate aminotransferase (AlAT, AsAT, alkaline phosphatase (ALP in serum, liver and heart homogenates. The degree of damage of the nervous system was studied by the activity of acetylcholinesterase (AChE in serum and liver homogenate. Content of TBA-AP in serum, liver and myocardium increased throughout the period of study. Reduction of TBA-AP was observed in studied tissues after applying of enterosgel sorbent in the affected body. After the defeat by toxicants we showed significant increase of fraction of AM1 (chain aminoacids dominate and AM2 (aromatic aminoacids dominate in serum and liver of experimental rats throughout the experiment. Enterosgel showed a positive effect on this index, reducing the content of MSM molecules. Due to toxic effects of endogenous and exogenous toxins in cells degradation and changing the permeability of plasma membranes of hepatocytes was observed as evidenced by the increased activity of both AlAT and As

  14. Simultaneous quantification of GABAergic 3alpha,5alpha/3alpha,5beta neuroactive steroids in human and rat serum.

    Science.gov (United States)

    Porcu, Patrizia; O'Buckley, Todd K; Alward, Sarah E; Marx, Christine E; Shampine, Lawrence J; Girdler, Susan S; Morrow, A Leslie

    2009-01-01

    The 3alpha,5alpha- and 3alpha,5beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone enhance GABAergic neurotransmission and produce inhibitory neurobehavioral and anti-inflammatory effects. Despite substantial information on the progesterone derivative (3alpha,5alpha)-3-hydroxypregnan-20-one (3alpha,5alpha-THP, allopregnanolone), the physiological significance of the other endogenous GABAergic neuroactive steroids has remained elusive. Here, we describe the validation of a method using gas chromatography-mass spectrometry to simultaneously identify serum levels of the eight 3alpha,5alpha- and 3alpha,5beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone. The method shows specificity, sensitivity and enhanced throughput compared to other methods already available for neuroactive steroid quantification. Administration of pregnenolone to rats and progesterone to women produced selective effects on the 3alpha,5alpha- and 3alpha,5beta-reduced neuroactive steroids, indicating differential regulation of their biosynthetic pathways. Pregnenolone administration increased serum levels of 3alpha,5alpha-THP (+1488%, psteroid was detected only in 3/16 control subjects. Levels of 3alpha,5alpha-A, 3alpha,5beta-A and pregnenolone were not altered. This method can be used to investigate the physiological and pathological role of neuroactive steroids and to develop biomarkers and new therapeutics for neurological and psychiatric disorders.

  15. Simultaneous Determination and Pharmacokinetic Study of Six Components in Rat Plasma by HPLC-MS/MS after Oral Administration of Acanthopanax sessiliflorus Fruit Extract

    OpenAIRE

    Peng Du; Mingdao Lei; Yu Liu; Shilin Yang

    2016-01-01

    A specific and reliable HPLC-MS/MS method was developed and validated for the simultaneous determination of protocatechuic acid (PCA), scopolin, (−)-pinoresinol-4,4′-di-O-β-d-glucopyranoside (PDG), acanthoside D, acanthoside B and hyperin in rat plasma for the first time. The analytes were separated on a C18 column (50 × 2.1 mm, 1.8 µm) and a triple-quadrupole mass spectrometer equipped with an electrospray ionization (ESI) source was used for detection. The rat plasma sample was prepared usi...

  16. Joint independent component analysis for simultaneous EEG-fMRI: principle and simulation.

    Science.gov (United States)

    Moosmann, Matthias; Eichele, Tom; Nordby, Helge; Hugdahl, Kenneth; Calhoun, Vince D

    2008-03-01

    An optimized scheme for the fusion of electroencephalography and event related potentials with functional magnetic resonance imaging (BOLD-fMRI) data should simultaneously assess all available electrophysiologic and hemodynamic information in a common data space. In doing so, it should be possible to identify features of latent neural sources whose trial-to-trial dynamics are jointly reflected in both modalities. We present a joint independent component analysis (jICA) model for analysis of simultaneous single trial EEG-fMRI measurements from multiple subjects. We outline the general idea underlying the jICA approach and present results from simulated data under realistic noise conditions. Our results indicate that this approach is a feasible and physiologically plausible data-driven way to achieve spatiotemporal mapping of event related responses in the human brain.

  17. FEASIBILITY ANALYSIS OF SAFETY SIMULTANEOUS EXTRACTION OF COAL AND COALBED METHANE

    Institute of Scientific and Technical Information of China (English)

    Li Shugang

    2000-01-01

    Coalbed methane is of great value to extract and utilize in China, but the result of such research is not satisfied yet today.The paper analyzed the storage characteristics of coalbed methane, and then studied the behavior of coalbed methane using the key stratum theory of strata control.According to the features related to coalbed methane accumulations and delivery, the technique for safely simultaneous extraction of coal and coalbed methane is proposed, and benefit analysis is made too.

  18. Simultaneous Elastic Recoil Detection Analysis of H and Other Elements in Foils

    Institute of Scientific and Technical Information of China (English)

    LU Xiu-Qin; ZHOU Ping; GUO Ji-Yu; ZHANG Xin; ZHAO Kui; NI Mei-Nan; SUI Li; MEI Jun-Ping; LIU Jian-Cheng

    2005-01-01

    @@ Hydrogen and other elements in SixNyHz foils have been simultaneously measured by using a single E(gas)- E(PSD) telescope and heavy 127I ion beam in elastic recoil detection analysis (ERDA). Hydrogen is measuredin the non-coincidence spectrum of E(PSD), and other elements from the △E - E coincidence spectrum. Thecomposition and depth profiling of the foils are obtained from the simulated spectra.

  19. Performance Analysis of the Microsoft Kinect Sensor for 2D Simultaneous Localization and Mapping (SLAM) Techniques

    OpenAIRE

    Kamarulzaman Kamarudin; Syed Muhammad Mamduh; Ali Yeon Md. Shakaff; Ammar Zakaria

    2014-01-01

    This paper presents a performance analysis of two open-source, laser scanner-based Simultaneous Localization and Mapping (SLAM) techniques (i.e., Gmapping and Hector SLAM) using a Microsoft Kinect to replace the laser sensor. Furthermore, the paper proposes a new system integration approach whereby a Linux virtual machine is used to run the open source SLAM algorithms. The experiments were conducted in two different environments; a small room with no features and a typical office corridor wi...

  20. Simultaneous Spectrophotometric Determination of Four Components including Acetaminophen by Taget Factor Analysis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    UV Spectrophotometric Target Factor Analysis (TFA) was used for the simultaneous determination of four components (acetaminophen, guuaifenesin, caffeine, Chlorphenamine maleate) in cough syrup. The computer program of TFA is based on VC++ language. The difficulty of overlapping of absorption spectra of four compounds was overcome by this procedure. The experimental results show that the average recovery of each component is all in the range from 98.9% to 106.8% and each component obtains satisfactory results without any pre-separation.

  1. Cytocompatible polyion complex gel of poly(Pro-Hyp-Gly) for simultaneous rat bone marrow stromal cell encapsulation.

    Science.gov (United States)

    Nurlidar, Farah; Kobayashi, Mime; Terada, Kayo; Ando, Tsuyoshi; Tanihara, Masao

    2017-10-01

    Polyion complex (PIC) gel of poly(Pro-Hyp-Gly) was successfully fabricated by simply mixing polyanion and polycation derivatives of poly(Pro-Hyp-Gly), a collagen-like polypeptide. The polyanion, succinylated poly(Pro-Hyp-Gly), and the polycation, arginylated poly(Pro-Hyp-Gly), contain carboxy (pKa = 5.2) and guanidinium (pKa = 12.4) groups, respectively. Mixing the polyanion and the polycation at physiological pH (pH = 7.4) resulted in PIC gel. The hydrogel formation was optimum at an equimolar ratio of carboxy to guanidinium groups, suggesting that ionic interaction is the main determinant for the hydrogel formation. The hydrogel was successfully used for simultaneous rat bone marrow stromal cell encapsulation. The encapsulated cells survived and proliferated within the hydrogel. In addition, the cells exhibited different morphology in the hydrogel compared with cells cultured on a tissue culture dish as a two-dimensional (2D) control. At day one, a round morphology and homogeneous single cell distribution were observed in the hydrogel. In contrast, the cells spread and formed a fibroblast-like morphology on the 2D control. After three days, the cells in the hydrogel maintained their morphology and some of them formed multicellular aggregates, which is similar to cell morphology in an in vivo microenvironment. These results suggest that the PIC gel of poly(Pro-Hyp-Gly) can serve as a cytocompatible three-dimensional scaffold for stem cell encapsulation, supporting their viability, proliferation, and in vivo-like behavior.

  2. Simultaneous isolation of DNA, RNA, and proteins for genetic, epigenetic, transcriptomic, and proteomic analysis.

    Science.gov (United States)

    Radpour, Ramin; Sikora, Michal; Grussenmeyer, Thomas; Kohler, Corina; Barekati, Zeinab; Holzgreve, Wolfgang; Lefkovits, Ivan; Zhong, Xiao Yan

    2009-11-01

    Analysis of DNA, RNA, and proteins for downstream genetic, epigenetic, transcriptomic, and proteomic analysis holds an important place in the field of medical care and life science. This is often hampered by the limited availability of sample material. For this reason, there exists an increasing interest for simultaneous isolation of DNA, RNA and proteins from a single sample aliquot. Several kit-systems allowing such a procedure have been introduced to the market. We present an approach using the AllPrep method for simultaneous isolation of DNA, RNA and proteins from several human specimens, such as whole blood, buffy coat, serum, plasma and tissue samples. The quantification and qualification of the isolated molecular species were assessed by different downstream methods: NanoDrop for measuring concentration and purity of all molecular species; DNA and RNA LabChip for fractionation analysis of nucleic acids; quantitative PCR for quantification analysis of DNA and RNA; thymidine-specific cleavage mass array on MALDI-TOF silico-chip for epigenetic analysis; Protein LabChip and two-dimensional (2D) gel electrophoresis for proteomic analysis. With our modified method, we can simultaneously isolate DNA, RNA and/or proteins from one single sample aliquot. We could overcome to some method limitations like low quality or DNA fragmentation using reamplification strategy for performing high-throughput downstream assays. Fast and easy performance of the procedure makes this method interesting for all fields of downstream analysis, especially when using limited sample resources. The cost-effectiveness of the procedure when material is abundantly available has not been addressed. This methodological improvement enables to execute such experiments that were not performable with standard procedure, and ensures reproducible outcome.

  3. An LC-MS/MS method for the simultaneous determination of 15 antipsychotics and two metabolites in hair and its application to rat hair.

    Science.gov (United States)

    Sim, Juhyun; Kim, Eunmi; Yang, Wonkyung; Woo, Sanghee; In, Sangwhan

    2017-05-01

    In recent years, the inappropriate use of antipsychotics by young Korean men has become a social problem. As military service exemptions are given for mental illness, some men pose as mental health patients to avoid military service. In order to verify the authenticity of mental illnesses, we developed simultaneous analytical methods for the detection of 15 antipsychotics and 2 of their metabolites in hair using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The target drugs were modafinil, atomoxetine, aripiprazole, benztropine, buspirone, duloxetine, gabapentin, oxcarbazepine, topiramate, escitalopram, paliperidone, ziprasidone, lamotrigine, clonazepam, levetiracetam, and metabolites of oxcarbazepine and clonazepam. To remove possible contaminants on the hair surface, hair samples were washed twice with methanol and distilled water, and then were extracted with methanol overnight at 38°C. Desipramine-d3 was used as an internal standard. LC-MS/MS analysis was performed on an Agilent 1290 Infinity UHPLC coupled to an AB Sciex Qtrap(®) 5500 MS/MS. The total chromatographic run time was 14min. The following validation parameters were evaluated: selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect, and recovery. The LOD and LOQ values for all analytes, except modafinil, ranged from 0.2 to 10pg/mg hair and from 0.2 to 20pg/mg hair, respectively. Good linearity was achieved for most of the analytes in the range of 20-200pg/mg hair. The method showed acceptable precision and accuracy, which were less than 15%, as well as satisfactory matrix effects and recoveries. Furthermore, this method was also applied to the analysis of rat hair samples. The study in rats showed that the concentrations of atomoxetine and aripiprazole in pigmented hair were significantly higher than those in non-pigmented hair. However, no significant difference was observed in the concentration of topiramate between

  4. Distributed analysis of simultaneous EEG-fMRI time-series: modeling and interpretation issues.

    Science.gov (United States)

    Esposito, Fabrizio; Aragri, Adriana; Piccoli, Tommaso; Tedeschi, Gioacchino; Goebel, Rainer; Di Salle, Francesco

    2009-10-01

    Functional magnetic resonance imaging (fMRI) and electroencephalography (EEG) represent brain activity in terms of a reliable anatomical localization and a detailed temporal evolution of neural signals. Simultaneous EEG-fMRI recordings offer the possibility to greatly enrich the significance and the interpretation of the single modality results because the same neural processes are observed from the same brain at the same time. Nonetheless, the different physical nature of the measured signals by the two techniques renders the coupling not always straightforward, especially in cognitive experiments where spatially localized and distributed effects coexist and evolve temporally at different temporal scales. The purpose of this article is to illustrate the combination of simultaneously recorded EEG and fMRI signals exploiting the principles of EEG distributed source modeling. We define a common source space for fMRI and EEG signal projection and gather a conceptually unique framework for the spatial and temporal comparative analysis. We illustrate this framework in a graded-load working-memory simultaneous EEG-fMRI experiment based on the n-back task where sustained load-dependent changes in the blood-oxygenation-level-dependent (BOLD) signals during continuous item memorization co-occur with parametric changes in the EEG theta power induced at each single item. In line with previous studies, we demonstrate on two single-subject cases how the presented approach is capable of colocalizing in midline frontal regions two phenomena simultaneously observed at different temporal scales, such as the sustained negative changes in BOLD activity and the parametric EEG theta synchronization. We discuss the presented approach in relation to modeling and interpretation issues typically arising in simultaneous EEG-fMRI studies.

  5. Cross-interference correction and simultaneous multi-gas analysis based on infrared absorption

    Institute of Scientific and Technical Information of China (English)

    Sun You-Wen; Zeng Yi; Liu Wen-Qing; Xie Pin-Hua; Chan Ka-Lok; Li Xian-Xin; Wang Shi-Mei; Huang Shu-Hua

    2012-01-01

    In this paper,we present simultaneous multiple pollutant gases (CO2,CO,and NO) measurements by using the non-dispersive infrared (NDIR) technique.A cross-correlation correction method is proposed and used to correct the cross-interferences among the target gases.The calculation of calibration curves is based on least-square fittings with third-order polynomials,and the interference functions are approximated by linear curves.The pure absorbance of each gas is obtained by solving three simultaneous equations using the fitted interference functions.Through the interference correction,the signal created at each filter channel only depends on the absorption of the intended gas.Gas mixture samples with different concentrations of CO2,CO,and NO are pumped into the sample cell for analysis.The results show that the measurement error of each gas is less than 4.5%.

  6. A UPLC-MS Method for Simultaneous Determination of Geniposidic Acid, Two Lignans and Phenolics in Rat Plasma and its Application to Pharmacokinetic Studies of Eucommia ulmoides Extract in Rats.

    Science.gov (United States)

    Li, Yongjun; Gong, Zipeng; Cao, Xu; Wang, Yonglin; Wang, Aimin; Zheng, Lin; Huang, Yong; Lan, Yanyu

    2016-10-01

    The bark of Eucommia ulmoides is a well-known Chinese herbal medicine that is used to regulate blood pressure and reduce blood sugar and fats, as well as an antioxidant and antimicrobial agent. Here we describe the development of a sensitive ultrahigh performance liquid chromatography-tandem mass spectrum method for the simultaneous determination of five major active ingredients of E. ulmoides bark extract, namely, geniposidic acid (GA), protocatechuic acid (PCA), chlorogenic acid (CA), (+)-pinoresinol di-O-β-D-glucopyranoside (PDG) and (+)-pinoresinol 4'-O-β-D-glucopyranoside (PG), in rat plasma. The preliminary steps in the plasma analysis were the addition of an internal standard and acidification (0.1 % formic acid), followed by protein precipitation with methanol. Separation of the active ingredients was performed on an ACQUITY UPLC® BEH C18 column (2.1 × 50 mm; internal diameter 1.7 µm) at a flow rate of 0.35 mL/min, with acetonitrile/water containing 0.1 % formic acid as the mobile phase. Detection was performed on a triple quadrupole tandem mass spectrometer via electrospray ionization source with positive and negative ionization modes. All calibration curves showed good linearity (r ≥ 0.997) over the concentration range with the low limit of quantification between 4.45 and 54.9 ng/mL. Precision was evaluated by intra- and inter-day assays, and the percentages of the relative standard deviation were all within 15 %. Extraction efficiency and matrix effect were 84.3-102.4 % and 98.1-112.2 %, respectively. The validated method was successfully applied to the pharmacokinetic study in rats after oral administration of E. ulmoides extract. The results indicate that the pharmacokinetic properties of GA differ from those of PCA, CA, PDG and PG, respectively.

  7. Dual phylogenetic staining protocol for simultaneous analysis of yeast and bacteria in artworks

    Science.gov (United States)

    González-Pérez, Marina; Brinco, Catarina; Vieira, Ricardo; Rosado, Tânia; Mauran, Guilhem; Pereira, António; Candeias, António; Caldeira, Ana Teresa

    2017-02-01

    The detection and analysis of metabolically active microorganisms are useful to determine those directly involved in the biodeterioration of cultural heritage (CH). Fluorescence in situ hybridization with oligonucleotide probes targeted at rRNA (RNA-FISH) has demonstrated to be a powerful tool for signaling them. However, more efforts are required for the technique to become a vital tool for the analysis of CH's microbiological communities. Simultaneous analysis of microorganisms belonging to different kingdoms, by RNA-FISH in-suspension approach, could represent an important progress: it could open the door for the future use of the technique to analyze the microbial communities by flow cytometry, which has shown to be a potent tool in environmental microbiology. Thus, in this work, various already implemented in-suspension RNA-FISH protocols for ex situ analysis of yeast and bacteria were investigated and adapted for allowing the simultaneous detection of these types of microorganisms. A deep investigation of the factors that can affect the results was carried out, focusing particular attention on the selection of the fluorochromes used for labelling the probe set. The resultant protocol, involving the use of EUK516-6-FAM/EUB338-Cy3 probes combination, was validated using artificial consortia and gave positive preliminary results when applied in samples from a real case study: the Paleolithic archaeological site of Escoural Cave (Alentejo, Portugal). This approach represents the first dual-staining RNA-FISH in-suspension protocol developed and applied for the simultaneous investigation of CH biodeteriogenic agents belonging to different kingdoms.

  8. Analysis of Heme oxygenase isomers in rat

    Institute of Scientific and Technical Information of China (English)

    Yun-ZhuLi; Wen-JunCui; Xue-HongZhang; Qing-XiangShen; JianWang; She

    2002-01-01

    AIM:To purify and identify heme oxygenase(HO) isomers which exist in rat liver,spleen and brain treated with hematin and phenylhydrazine and in untrated rat liver and to investigate the characteristics of HO isomers,to isolate and confirm the rat HO-1 cDNA that actually encodes HO-1 by expressing cDNA in monkey Kidney cells(COS-1 cells),to prepare the rat heme oxygenase-1(HO-1)mutant and to detect inhibition of HO-1 mutated enzyme.

  9. Simultaneous detection of antibodies to five Actinobacillus pleuropneumoniae serovars using bead-based multiplex analysis

    DEFF Research Database (Denmark)

    Berger, Sanne Schou; Lauritsen, Klara Tølbøl; Boas, Ulrik

    2017-01-01

    We have developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested...... Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC...

  10. Simultaneous Impedance Analysis of Three Parallel Piezoelectric Quartz Crystals for Electrochemical Depletion Layer Effect Study

    Institute of Scientific and Technical Information of China (English)

    Hui Yan liu; Qing Ji XIE

    2004-01-01

    Simultaneous impedance analysis of three one-face sealed resonating piezoelectric quartz crystals (PQCs) in parallel is proposed through admittance measurements of the three PQCs on one impedance analyzer and then non-linear fitting according to the parallel combination of three Butterworth-Van Dyke circuits. Responses of each PQC obtained from the three-PQC mode agreed well with those measured separately in series sucrose aqueous solutions. This novel method has been used for the study of depletion-layer effect during ferri-/ferrocyanide electrochemical reactions.

  11. Determining the cation exchange capacity of montmorillonite by simultaneous thermal analysis method

    Science.gov (United States)

    Boeva, N. M.; Bocharnikova, Yu. I.; Belousov, P. E.; Zhigarev, V. V.

    2016-08-01

    A way of determining the cation exchange capacity of montmorillonite by simultaneous thermal analysis is developed using as an example the bentonites of the 10th Khutor deposit (Republic of Khakassia) and the Vodopadnyi area (Sakhalin Island). A correlation is established between the cation exchange capacity of smectite and its weight loss upon heating in the range of dehydration; the enthalpy of dehydration of montmorillonite; and the weight loss and the enthalpy of thermal dissociation of ethylene glycol contained in the interlayer space of the mineral's crystal structure. These data open up new possibilities for determining the cation exchange capacity of montmorillonite, the most important technological indicator of the natural clay nanomineral.

  12. Thermodynamic investigations of nitroxoline sublimation by simultaneous DSC-FTIR method and isothermal TG analysis.

    Science.gov (United States)

    Gao, Gau-Yi; Lin, Shan-Yang

    2010-01-01

    To investigate the physicochemical characteristics, thermodynamics, possible sublimation process and kinetics of nitroxoline, differential scanning calorimetry (DSC), isothermal thermogravimetry (TG), and Fourier transform infrared (FTIR) microspectroscopy equipped with a micro hot-stage of DSC microscopy assembly (simultaneous DSC-FTIR method) were used. The DSC result indicates that nitroxoline exhibited a sharp endothermic peak at 182 degrees C with enthalpy of 103.1 J/g due to the melting point of nitroxoline. A sublimation behavior of nitroxoline was found from 129 degrees C by gradual weight loss in TG curve. However, the nonisothermal DSC-FTIR method reveals that the temperature at 95 degrees C was the onset temperature of nitroxoline sublimation. A significant difference between DSC-FTIR method and TG analysis suggests that the simultaneous DSC-FTIR method was more sensitive than that of the TG analysis to detect the beginning temperature of nitroxoline sublimation. The sublimation kinetics of nitroxoline determined by isothermal TG analysis evidenced that the zero-order kinetics was followed over the sublimation time. The sublimation enthalpy correction was also carried out by a group additivity approach for the estimation of heat capacity. The enthalpy of nitroxoline sublimation estimated was 86.14 KJ/mol at 298.15 K.

  13. Simultaneous HPLC quantitative analysis of active compounds in leaves of Moringa oleifera Lam.

    Science.gov (United States)

    Vongsak, Boonyadist; Sithisarn, Pongtip; Gritsanapan, Wandee

    2014-08-01

    Moringa oleifera Lam. has been used as a traditional medicine for the treatment of numerous diseases. A simultaneous high-performance liquid chromatography (HPLC) analysis was developed and validated for the determination of the contents of crypto-chlorogenic acid, isoquercetin and astragalin, the primary antioxidative compounds, in M. oleifera leaves. HPLC analysis was successfully conducted by using a Hypersil BDS C18 column, eluted with a gradient of methanol-1% acetic acid with a flow rate of 1 mL/min, and detected at 334 nm. Parameters for the validation included linearity, precision, accuracy and limits of detection and quantitation. The developed HPLC method was precise, with relative standard deviation oleifera leaf extracts were 98.50, 98.47 and 98.59%, respectively. The average contents of these compounds in the dried ethanolic extracts of the leaves of M. oleifera collected from different regions of Thailand were 0.081, 0.120 and 0.153% (w/w), respectively. The developed HPLC method was appropriate and practical for the simultaneous analysis of crypto-chlorogenic acid, isoquercetin and astragalin in the leaf extract of M. oleifera. This work is valuable as guidance for the standardization of the leaf extracts and pharmaceutical products of M. oleifera. © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  14. A microfluidic cigarette smoke collecting platform for simultaneous sample extraction and multiplex analysis.

    Science.gov (United States)

    Hu, Shan-Wen; Xu, Bi-Yi; Qiao, Shu; Zhao, Ge; Xu, Jing-Juan; Chen, Hong-Yuan; Xie, Fu-Wei

    2016-04-01

    In this work, we report a novel microfluidic gas collecting platform aiming at simultaneous sample extraction and multiplex mass spectrometry (MS) analysis. An alveolar-mimicking elastic polydimethylsiloxane (PDMS) structures was designed to move dynamically driven by external pressure. The movement was well tuned both by its amplitude and rhythm following the natural process of human respiration. By integrating the alveolar units into arrays and assembling them to gas channels, a cyclic contraction/expansion system for gas inhale and exhale was successfully constructed. Upon equipping this system with a droplet array on the alveolar array surface, we were able to get information of inhaled smoke in a new strategy. Here, with cigarette smoke as an example, analysis of accumulation for target molecules during passive smoking is taken. Relationships between the breathing times, distances away from smokers and inhaled content of nicotine are clarified. Further, by applying different types of extraction solvent droplets on different locations of the droplet array, simultaneous extraction of nicotine, formaldehyde and caproic acid in sidestream smoke (SS) are realized. Since the extract droplets are spatially separated, they can be directly analyzed by MS which is fast and can rid us of all complex sample separation and purification steps. Combining all these merits, this small, cheap and portable platform might find wide application in inhaled air pollutant analysis both in and outdoors. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Simultaneous Determination of Gold and Platinum by Double Artificial Neural Network Analysis with Flow-injection Chemiluminescence

    Institute of Scientific and Technical Information of China (English)

    Ming Yang LIU; Hai Tao ZHANG; Jun Feng LI; Shu Gui CHEN; Hong Yan WANG

    2006-01-01

    A highly sensitive double artificial neural network (DANN) analysis with flow-injection chemiluminescence (FI-CL) has been developed to simultaneously determine the trace amounts of the gold and platinum in simulated mixed samples, without the boring process.

  16. Simultaneous measurement of kidney function by dynamic contrast enhanced MRI and FITC-sinistrin clearance in rats at 3 tesla: initial results.

    Directory of Open Access Journals (Sweden)

    Frank G Zöllner

    Full Text Available Glomerular filtration rate (GFR is an essential parameter of kidney function which can be measured by dynamic contrast enhanced magnetic resonance imaging (MRI-GFR and transcutaneous approaches based on fluorescent tracer molecules (optical-GFR. In an initial study comparing both techniques in separate measurements on the same animal, the correlation of the obtained GFR was poor. The goal of this study was to investigate if a simultaneous measurement was feasible and if thereby, the discrepancies in MRI-GFR and optical-GFR could be reduced. For the experiments healthy and unilateral nephrectomised (UNX Sprague Dawley (SD rats were used. The miniaturized fluorescent sensor was fixed on the depilated back of an anesthetized rat. A bolus of 5 mg/100 g b.w. of FITC-sinistrin was intravenously injected. For dynamic contrast enhanced perfusion imaging (DCE-MRI a 3D time-resolved angiography with stochastic trajectories (TWIST sequence was used. By means of a one compartment model the excretion half-life (t1/2 of FITC-sinistrin was calculated and converted into GFR. GFR from DCE-MRI was calculated by fitting pixel-wise a two compartment renal filtration model. Mean cortical GFR and GFR by FITC-sinistrin were compared by Bland-Altman plots and pair-wise t-test. Results show that a simultaneous GFR measurement using both techniques is feasible. Mean optical-GFR was 4.34 ± 2.22 ml/min (healthy SD rats and 2.34 ± 0.90 ml/min (UNX rats whereas MRI-GFR was 2.10 ± 0.64 ml/min (SD rats and 1.17 ± 0.38 ml/min (UNX rats. Differences between healthy and UNX rats were significant (p<0.05 and almost equal percentage difference (46.1% and 44.3% in mean GFR were assessed with both techniques. Overall mean optical-GFR values were approximately twice as high compared to MRI-GFR values. However, compared to a previous study, our results showed a higher agreement. In conclusion, the possibility to use the transcutaneous method in MRI may have a huge impact in

  17. Simultaneous quantification of 5 main components of Psoralea corylifolia L. in rats' plasma by utilizing ultra high pressure liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Gao, Qianqian; Xu, Zisheng; Zhao, Genhua; Wang, Heng; Weng, Zebin; Pei, Ke; Wu, Li; Cai, Baochang; Chen, Zhipeng; Li, Weidong

    2016-02-01

    Psoralea corylifolia L. has long been used in traditional Chinese medicine for treating and preventing many diseases. A group of flavonoid components are regarded as the active principals within the seeds. In this research, a rapid, accurate and sensitive ultra high pressure liquid chromatography tandem mass spectrometry (UHPLC/MS/MS) method has been established for simultaneous quantification of its 5 main components, namely, neobavaisoflavone, bavachin, isobavachalcone, bavachinin and corylifol A in rats' plasma after the rats were orally administrated with Buguzhi extract. Negative ion electrospray mode was applied in the detection process. Multiple reactions monitoring (MRM) mode was utilized for simultaneous quantitative analyzing of neobavaisoflavone (m/z 321.1→m/z 265.1), bavachin (m/z 323.1→m/z 119.0), isobavachalcone (m/z 323.2→m/z 119.0), bavachinin (m/z 337.2→m/z 119.0), corylifol A (m/z 389.2→m/z 277.0) and liquiritigenin (Internal Standard, m/z 255.1→m/z 119.0). Chromatographic separation of the above mentioned components was conducted on a Waters BEH-C18 column (100 mm×2.1mm, 1.7μm) with gradient elution system at flow rate of 0.3mL/min. The mobile phase was composed of acetonitrile and 0.1% formic acid solution. The lower limit of quantification (LLOQ) for each of the above analytes was 0.5ng/mL. Each of the analytes exhibited good linearity within the concentration range of 0.5-100ng/mL. The method was fully validated for its selectivity, accuracy, precision, stability, matrix effect and extraction recovery. The validated method has been successfully applied for simultaneous determination of the 5 flavonoids in rat plasma for the first time.

  18. Simultaneous determination of ascaridole, p-cymene and α-terpinene in rat plasma after oral administration of Chenopodium ambrosioides L. by GC-MS.

    Science.gov (United States)

    Hu, Xiaoqian; Chu, Yang; Ma, Gang; Li, Wei; Wang, Xiangyang; Mo, Hongmei; Yin, Qihui; Guo, Jiahua; Ma, Xiaohui; Zhou, Shuiping

    2015-11-01

    A sensitive and reliable GC-MS method was developed and validated for the simultaneous determination of ascaridole, p-cymene and α-terpinene in rat plasma using naphthalene as internal standard. The plasma samples were extracted with ethyl acetate. Chromatographic separation was carried out on a HP-5MS capillary analytical column (30 m × 0.25 mm, 0.25 µm) and detection was performed on a quadrupole mass spectrometer detector operated under selected ion monitoring mode. The method showed excellent linearity over the investigated concentration range (r > 0.99) with the limit of quantitation down to 50, 10 and 5 ng/mL for ascaridole, p-cymene and α-terpinene, respectively. The intra-day and inter-day precisions (RSD) were Chenopodium ambrosioides L. following oral administration to rats. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Simultaneous effects on parvalbumin-positive interneuron and dopaminergic system development in a transgenic rat model for sporadic schizophrenia

    Science.gov (United States)

    Hamburg, Hannah; Trossbach, Svenja V.; Bader, Verian; Chwiesko, Caroline; Kipar, Anja; Sauvage, Magdalena; Crum, William R.; Vernon, Anthony C.; Bidmon, Hans J.; Korth, Carsten

    2016-01-01

    To date, unequivocal neuroanatomical features have been demonstrated neither for sporadic nor for familial schizophrenia. Here, we investigated the neuroanatomical changes in a transgenic rat model for a subset of sporadic chronic mental illness (CMI), which modestly overexpresses human full-length, non-mutant Disrupted-in-Schizophrenia 1 (DISC1), and for which aberrant dopamine homeostasis consistent with some schizophrenia phenotypes has previously been reported. Neuroanatomical analysis revealed a reduced density of dopaminergic neurons in the substantia nigra and reduced dopaminergic fibres in the striatum. Parvalbumin-positive interneuron occurrence in the somatosensory cortex was shifted from layers II/III to V/VI, and the number of calbindin-positive interneurons was slightly decreased. Reduced corpus callosum thickness confirmed trend-level observations from in vivo MRI and voxel-wise tensor based morphometry. These neuroanatomical changes help explain functional phenotypes of this animal model, some of which resemble changes observed in human schizophrenia post mortem brain tissues. Our findings also demonstrate how a single molecular factor, DISC1 overexpression or misassembly, can account for a variety of seemingly unrelated morphological phenotypes and thus provides a possible unifying explanation for similar findings observed in sporadic schizophrenia patients. Our anatomical investigation of a defined model for sporadic mental illness enables a clearer definition of neuroanatomical changes associated with subsets of human sporadic schizophrenia. PMID:27721451

  20. Improved simultaneous determination method of beta-carotene and retinol with saponification in human serum and rat liver.

    Science.gov (United States)

    Hosotani, Keisuke; Kitagawa, Masahiro

    2003-07-01

    Among the many simultaneous determination methods for carotenoid and retinoid, there are only a few reports including the saponification process. However, the yields of beta-carotene and retinol were higher when using this process. In this study, the analytical conditions, including saponification, were investigated. The extraction solvent was n-hexane and the sample solvent was HPLC mobile phase in the beta-carotene and retinol analysis. BHT as an antioxidant was added at concentrations of 0.125 and 0.025%, respectively, to ethanol and n-hexane phase in the extraction process for serum. The recovery rates were 99.7, 93.7 and 98.3% for beta-carotene, retinol and retinyl palmitate in serum, respectively, and 107.1, 92.8 and 98.8% for beta-carotene, retinol and retinyl palmitate in liver, respectively. The within-day coefficients of variation (C.V.) were 6.0% for serum and 4.7% for liver in the case of beta-carotene, 7.1% for serum, and 5.1% for liver in the case of retinol. The between-day coefficients of variation were 2.7% for serum and 2.7% for liver in the case of beta-carotene, and for retinol, 6.4% for serum and 2.7% for liver.

  1. a Psycholinguistic Model for Simultaneous Translation, and Proficiency Assessment by Automated Acoustic Analysis of Discourse.

    Science.gov (United States)

    Yaghi, Hussein M.

    Two separate but related issues are addressed: how simultaneous translation (ST) works on a cognitive level and how such translation can be objectively assessed. Both of these issues are discussed in the light of qualitative and quantitative analyses of a large corpus of recordings of ST and shadowing. The proposed ST model utilises knowledge derived from a discourse analysis of the data, many accepted facts in the psychology tradition, and evidence from controlled experiments that are carried out here. This model has three advantages: (i) it is based on analyses of extended spontaneous speech rather than word-, syllable-, or clause -bound stimuli; (ii) it draws equally on linguistic and psychological knowledge; and (iii) it adopts a non-traditional view of language called 'the linguistic construction of reality'. The discourse-based knowledge is also used to develop three computerised systems for the assessment of simultaneous translation: one is a semi-automated system that treats the content of the translation; and two are fully automated, one of which is based on the time structure of the acoustic signals whilst the other is based on their cross-correlation. For each system, several parameters of performance are identified, and they are correlated with assessments rendered by the traditional, subjective, qualitative method. Using signal processing techniques, the acoustic analysis of discourse leads to the conclusion that quality in simultaneous translation can be assessed quantitatively with varying degrees of automation. It identifies as measures of performance (i) three content-based standards; (ii) four time management parameters that reflect the influence of the source on the target language time structure; and (iii) two types of acoustical signal coherence. Proficiency in ST is shown to be directly related to coherence and speech rate but inversely related to omission and delay. High proficiency is associated with a high degree of simultaneity and

  2. Simultaneous immunoblotting analysis with activity gel electrophoresis and 2-D gel electrophoresis.

    Science.gov (United States)

    Lee, Der-Yen; Chang, Geen-Dong

    2015-01-01

    Diffusion blotting method can couple immunoblotting analysis with another biochemical technique in a single polyacrylamide gel, however, with lower transfer efficiency as compared to the conventional electroblotting method. Thus, with diffusion blotting, protein blots can be obtained from an SDS polyacrylamide gel for zymography assay, from a native polyacrylamide gel for electrophoretic mobility shift assay (EMSA) or from a 2-D polyacrylamide gel for large-scale screening and identification of a protein marker. Thereafter, a particular signal in zymography, electrophoretic mobility shift assay, and 2-dimensional gel can be confirmed or identified by simultaneous immunoblotting analysis with a corresponding antiserum. These advantages make diffusion blotting desirable when partial loss of transfer efficiency can be tolerated or be compensated by a more sensitive immunodetection reaction using enhanced chemiluminescence detection.

  3. Simultaneous Greedy Analysis Pursuit for compressive sensing of multi-channel ECG signals.

    Science.gov (United States)

    Avonds, Yurrit; Liu, Yipeng; Van Huffel, Sabine

    2014-01-01

    This paper addresses compressive sensing for multi-channel ECG. Compared to the traditional sparse signal recovery approach which decomposes the signal into the product of a dictionary and a sparse vector, the recently developed cosparse approach exploits sparsity of the product of an analysis matrix and the original signal. We apply the cosparse Greedy Analysis Pursuit (GAP) algorithm for compressive sensing of ECG signals. Moreover, to reduce processing time, classical signal-channel GAP is generalized to the multi-channel GAP algorithm, which simultaneously reconstructs multiple signals with similar support. Numerical experiments show that the proposed method outperforms the classical sparse multi-channel greedy algorithms in terms of accuracy and the single-channel cosparse approach in terms of processing speed.

  4. Simultaneous determination and pharmacokinetics of five alkaloids in rat plasma by ultra high performance liquid chromatography with tandem mass spectrometry after the oral administration of Corydalis bungeana Turcz extract.

    Science.gov (United States)

    Liu, Lu; Yu, Xueli; Wang, Zhibin; Sun, Jiahui; Pan, Rong; Yang, Chunjuan; Wu, Lijun

    2016-01-01

    Pharmacokinetic studies were conducted on rats for protopine, corynoline, 7'-(3',4'-dihydroxyphenyl)-N-[(4-methoxyphenyl)-ethyl]propenamide, acetylcorynoline, and 8-oxocorynoline, five main active components from Corydalis bungeana Turcz (C. bungeana Turcz). An ultra high performance liquid chromatography with tandem mass spectrometry method has been developed and validated for the simultaneous determination of the pharmacokinetic characteristics of these components in rat plasma. Chromatographic separation was achieved on an Agilent SB-C18 column (1.8 μm, 150 × 2.1 mm) using a gradient elution program with a mobile phase consisting of acetonitrile and water containing 0.1% acetic acid at a flow rate of 0.3 mL/min. The analytes were detected in the multiple reaction monitoring mode with positive electrospray ionization. Lower limits of quantification were >0.680 ng/mL and matrix effects ranged from 91.26 to 100.38%. The mean extraction recoveries of quality control samples were less than 79.32%, and the precision and accuracy were within the acceptable limits. All analytes were proven to be stable during sample storage and analysis procedures. The validated method was successfully applied to the pharmacokinetic study of five alkaloid components after oral administration of C. bungeana Turcz extract to rats. The obtained results may be helpful to reveal the mechanism of action and to guide the clinical application of C. bungeana Turcz.

  5. Simultaneous determination of metoprolol, propranolol and phenol red in samples from rat in situ intestinal perfusion studies

    Directory of Open Access Journals (Sweden)

    Parvin Zakeri-Milani

    2006-05-01

    Full Text Available Single-pass intestinal perfusion technique (SPIP is the most used classic technique employed in the study of intestinal absorption of compounds in which a non-absorbable marker such as phenol red is used to correct the water flux. A simple and rapid reversed-phase high performance liquid chromatographic method with UV detection at 227 nm was developed for simultaneous quantitation of propranolol and metoprolol along with phenol red for in-situ permeability studies. The mobile phase was a mixture of 55% methanol, 45% of 0.05 M KH2PO4 aqueous solution (adjusted to pH 6 and 0.2 % (v/v triethylamine. Analysis was run at a flow rate of 1 ml/min with a 9 min run time. The calibration curves were linear for all three compounds (r > 0.999 across the concentration range of 7.5-125 μg/ml with a limit of detection of 4.24, 2.18 and 8.57 ng/ml and limit of quantification of 14, 7.2 and 28.3 ng/ml for metoprolol, propranolol and phenol red respectively. The coefficient of variation for intra-assay and inter-assay precision was less than 8% and the accuracy was between 93.6-107%. Using the SPIP technique and the suggested HPLC method for sample analysis, the mean values of 0.49 e-4 (±0.19 cm/sec and 0.32 e-4 (± 0.09 cm/sec were obtained for propranolol and metoprolol intestinal permeability coefficients respectively.

  6. Simultaneous RNA-seq analysis of a mixed transcriptome of rice and blast fungus interaction.

    Directory of Open Access Journals (Sweden)

    Yoshihiro Kawahara

    mixed transcriptome analysis is useful for the simultaneous elucidation of the tactics of host plant defense and pathogen attack.

  7. Further Enrichment and Analysis of Rat CFU-s

    Science.gov (United States)

    1990-01-01

    argon laser was tuned to the UV and was used to excite the DAPI-DNA complex. By gating on the APC- Ox720 and PhyB-x22- cells, a new FITC-W3/13...Thy-I and light scatter properties of CFU-s from cortisone pre-treated rats. The cortisone treatment diminished marrow lymphocytes and separated them...from cortisone - resistant CFU-s. Starting with marrow from normal rats, Goldschneider et al. Further Enrichment and Analysis of Rat CFU-s 190 A

  8. Simultaneous analysis method for GHB, ketamine, norketamine, phenobarbital, thiopental, zolpidem, zopiclone and phenytoin in urine, using C18 poroshell column.

    Science.gov (United States)

    Anilanmert, Beril; Çavuş, Fatma; Narin, Ibrahim; Cengiz, Salih; Sertler, Şefika; Özdemir, Ali Acar; Açikkol, Münevver

    2016-06-01

    Date-rape drugs have the potential to be used in drug-facilitated sexual assault, organ theft and property theft. Since they are colorless, tasteless and odorless, victims can drink without noticing, when added to the beverages. These drugs must be detected in time, before they are cleared up from the biofluids. A simultaneous extraction and determination method in urine for GHB, ketamine, norketamine, phenobarbital, thiopental, zolpidem, zopiclone and phenytoin (an anticonvulsant and antiepileptic drug) with LC-MS/MS was developed for the first time with analytically acceptable recoveries and validated. A 4 steps liquid-liquid extraction was applied, using only 1.000mL urine. A new age commercial C18 poroshell column with high column efficiency was used for LC-MS/MS analysis with a fast isocratic elution as 5.5min. A new MS transition were introduced for barbital. 222.7>179.8 with the effect of acetonitrile. Recoveries (%) were between 80.98-99.27 for all analytes, except for GHB which was 71.46. LOD and LOQ values were found in the ranges of 0.59-49.50 and 9.20-80.80ngmL(-1) for all the analytes (except for GHB:3.44 and 6.00μgmL(-1)). HorRat values calculated (between 0.25-1.21), revealed that the inter-day and interanalist precisions (RSD%≤14.54%) acceptable. The simultaneous extraction and determination of these 8 analytes in urine is challenging because of the difficulty arising from the different chemical properties of some. Since the procedure can extract drugs from a wide range of polarity and pKa, it increases the window of detection. Group representatives from barbiturates, z-drugs, ketamine, phenytoin and polar acidic drugs (GHB) have been successfully analyzed in this study with low detection limits. The method is important from the point of determining the combined or single use of these drugs in crimes and finding out the reasons of deaths related to these drugs.

  9. Proteomic analysis of hippocampus in the rat

    Institute of Scientific and Technical Information of China (English)

    ZHANG Bo; WANG Ren-zhi; LIAN Zhi-gang; YAO Yong

    2004-01-01

    Objective To analyze the protein expression in the rat hippocampus by the proteomic approach.Methods Proteins from hippocampal tissue homogenates of the rat were separated by two-dimensional gel electrophoresis(2-DE),and stained with colloidal Coomassie blue to produce a high-resolution map of the rat hippocampus proteome.Selected proteins from this map were digested with trypsin,and the resulting tryptic peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).The mass spectrometric data were used to identify the proteins through searches of the NCBI protein sequence database.Results 37 prominent proteins with various functional characteristics were identified.The identified brain protein classes covered metabolism enzymes,cytoskeleton proteins,heat shock proteins,antioxidant proteins,signalling proteins,proteasome-related proteins,neuron-specific proteins and glial-associated proteins.Furthermore,3 hypothetical proteins,unknown proteins so far only proposed from their nucleic acid structure,were identified.Conclusion This study provides the first unbiased characterization of proteins of the rat hippocampus and will be used for future studies of differential protein expression in rat models of neurological disorders.

  10. Simultaneous analysis of saturated and unsaturated fatty acids present in pequi fruits by capillary electrophoresis

    Directory of Open Access Journals (Sweden)

    Patrícia M. de Castro Barra

    2013-01-01

    Full Text Available In the current study, an alternative method has been proposed for simultaneous analysis of palmitic, stearic, oleic, linoleic, and linolenic acids by capillary zone electrophoresis (CZE using indirect detection. The background electrolyte (BGE used for the analysis of these fatty acids (FAs consisted of 15.0 mmol L−1 NaH2PO4/Na2HPO4 at pH 6.86, 4.0 mmol L−1 SDBS, 8.3 mmol L−1 Brij 35, 45% v/v acetonitrile (can, and 2.1% n-octanol. The FAs quantification of FAs was performed using a response factor approach, which provided a high analytical throughput for the real sample. The CZE method, which was applied successfully for the analysis of pequi pulp, has advantages such as short analysis time, absence of lipid fraction extraction and derivatization steps, and no significant difference in the 95% confidence intervals for FA quantification results, compared to the gas chromatography official method (AOCS Ce 1h-05.

  11. HPTLC method for simultaneous analysis of stevioside and rebaudioside-A in Stevia rebaudiana.

    Science.gov (United States)

    Londhe, Smita V; Nanaware, Sadhana M

    2013-01-01

    An HPTLC method was developed and validated for simultaneous determination of stevioside (STE) and rebaudioside-A (REB-A) in Stevia rebaudiana leaves. The HPTLC separation was performed on precoated silica gel 60F254 HPTLC plates with the mobile phase ethyl acetate-ethanol-acetone-water (15 + 3 + 6 + 6, v/v/v/v). The densitometric quantification of steviol glycosides was carried out at Amax 580 nm in the reflection-absorption mode after spraying with anisaldehyde-sulfuric acid reagent. Rf values were 0.34 and 0.28 for STE and REB-A, respectively. The content of STE and REB-A in leaf extract was found to be 6.94 and 6.35%, respectively. The method was validated in terms of precision, accuracy, specificity, and robustness. The method can be useful for routine analysis of the sweeteners.

  12. Simultaneous Analysis of Multiple Promoters: An Application of the PC-GW Binary Vector Series.

    Science.gov (United States)

    Dalal, Jyoti

    2016-01-01

    With the advances in the field of synthetic biology, there is an increasing demand for multi-gene cloning technologies. Molecular cloning to generate multi-gene constructs can be performed by restriction digestion, or by recombination-based cloning strategies such as Gateway(®). This chapter details cloning, transformation, and selection procedures involved in generation of multi-gene expressing transgenic plants. Methods are described for cloning five distinct promoter-reporter fusion constructs into the PC-GW-BAR vector (from the PC-GW vector series) using Gateway(®) technology and meganuclease sites. Further, transformation and selection methods are described for the biofuel crop Camelina sativa from the Brassicaceae family. These methods would be constructive toward generating multi-gene expressing plants for simultaneous expression analysis of five promoters in a short time period.

  13. Digital TV-echelle spectrograph for simultaneous multielemental analysis using microcomputer control

    Energy Technology Data Exchange (ETDEWEB)

    Davidson, J.B.; Case, A.L.

    1980-12-01

    A digital TV-echelle spectrograph with microcomputer control was developed for simultaneous multielemental analysis. The optical system is a commercially available unit originally equipped for film and photomultiplier (single element) readout. The film port was adapted for the intensifier camera. The camera output is digitized and stored in a microcomputer-controlled, 512 x 512 x 12 bit memory and image processor. Multiple spectra over the range of 200 to 800 nm are recorded in a single exposure. Spectra lasting from nanoseconds to seconds are digitized and stored in 0.033 s and displayed on a TV monitor. An inexpensive microcomputer controls the exposure, reads and displays the intensity of predetermined spectral lines, and calculates wavelengths of unknown lines. The digital addresses of unknown lines are determined by superimposing a cursor on the TV display. The microcomputer also writes into memory wavelength fiducial marks for alignment of the TV camera.

  14. Continuous Fourier transform method and apparatus. [for the analysis of simultaneous analog signal components

    Science.gov (United States)

    Munoz, R. M. (Inventor)

    1974-01-01

    An input analog signal to be frequency analyzed is separated into N number of simultaneous analog signal components each identical to the original but delayed relative to the original by a successively larger time delay. The separated and delayed analog components are combined together in a suitable number of adders and attenuators in accordance with at least one component product of the continuous Fourier transform and analog signal matrices to separate the analog input signal into at least one of its continuous analog frequency components of bandwidth 1/N times the bandwidth of the original input signal. The original analog input signal can be reconstituted by combining the separate analog frequency components in accordance with the component products of the continuous Fourier transform and analog frequency component matrices. The continuous Fourier transformation is useful for spectrum analysis, filtering, transfer function synthesis, and communications.

  15. Simultaneous determination of some common food dyes in commercial products by digital image analysis

    Directory of Open Access Journals (Sweden)

    Mohammad-Hossein Sorouraddin

    2015-09-01

    Full Text Available A simple and relatively fast image-analysis method using digital images, obtained with a flatbed scanner, has been described. The method was used for the simultaneous determination of four common food dyes, namely, carmoisine, brilliant blue, sunset yellow, and quinoline yellow, in binary mixtures in commercial products without a need for any prior separation steps. The results obtained were validated against a standard high-performance liquid chromatography method and a good agreement was obtained. The parameters affecting the experimental results were optimized. Under the optimal conditions, the method provided acceptable linear ranges (20–250 mg/L with correlation coefficients higher than 0.998, suitable precision (relative standard deviation ≤ 4.5%, and limits of detection between 4.82 and 8.05 mg/L.

  16. Method for simultaneous analysis of eight analogues of vitamin D using liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Shah Iltaf

    2012-10-01

    Full Text Available Abstract Background Despite considerable global investigation over several decades, the roles of vitamin D in health and disease development remains convoluted. One recognised issue is the difficulty of accurately measuring the active forms of vitamin D. Advances made include some new methods addressing the potential interference by excluding epimers and isobars. However, there is no evidence that epimers are without function. Therefore, the aim of this study was to develop and validate, for the first time, a new assay to simultaneously measure levels of six forms of vitamin D along with two epimers. The assay was applied to multilevel certified reference material (CRM and 25 pooled human sera samples, obtained from the Vitamin D External Quality Assessment Scheme (DEQAS, to demonstrate its efficiency. Results The assay is capable of simultaneously measuring eight vitamin D analogues over the calibration ranges and LODs (in nmol/L of: 1α25(OH2D2 [0.015-1; 0.01], 1α25(OH2D3 [0.1-100; 0.01], 25OHD3 [0.5-100, 0.025], 3-epi-25OHD3 [0.1-100, 0.05], 25OHD2 [0.5-100, 0.025], 3-epi-25OHD2 [0.1-100, 0.05], vitamin D3 [0.5-100, 0.05] and vitamin D2 [0.5-100, 0.05], using stanozolol-d3 as internal standard. Certified reference material and external quality control samples (DEQAS were analysed to meet the standards outlined by National Institute of Standards and Technology (NIST. Validation steps included recovery and both precision and accuracy under inter- and intra-day variation limit of detection, and analysis of each analyte over a linear range. All validation parameters were in line with acceptable Food and Drug Administration (FDA guidelines. All eight analogues were quantified with the 25OHD levels being commensurate with DEQAS data. Conclusions This report details the application of a new LC-MS/MS based assay for the efficient analysis of eight analogues of vitamin D over a range of samples, which is a significant advance over the existing

  17. Human saliva cortisone and cortisol simultaneous analysis using reverse phase HPLC technique.

    Science.gov (United States)

    De Palo, Elio F; Antonelli, Giorgia; Benetazzo, Arianna; Prearo, Maddalena; Gatti, Rosalba

    2009-07-01

    Hyper-hypo tension (like Cushing's syndrome, apparent mineralocorticoid excess syndrome and Addison's disease) diagnostic laboratory requires cortisol (F) analysis. The simultaneous analysis of human saliva F and cortisone (E), the inactive F metabolite, by solid phase extraction and RP-HPLC was studied. Saliva/standard samples were C18-SPE extracted, dried and resuspended. E and F were analysed by isocratic RP-HPLC (acetonitrile/water 27/73%) and UV detection. In the morning and in the evening Salivette stimulated saliva specimens were collected from healthy volunteers. The E and F calibration curve ranges were 11.0-110.0 and 5.5-55.0 nmol/l respectively. The LOD was 0.2 and 0.1 nmol/l for E and F respectively. The intra and inter assay CVs were respectively 2.7-6.6 and 5.6-7.0% for E and 5.8-7.0 and 11.7-13.1% for F. The E and F spiked saliva sample recovery was 99% and 88% respectively. Saliva specimen stability was validated. E and F saliva levels in healthy volunteers were significantly (p<0.001) higher at 8 a.m. compared with 11 p.m. (26.4+/-8.9 vs. 4.3+/-2.9 nmol/l for E; 11.1+/-4.0 vs. 2.5+/-1.5 nmol/l for F, respectively). This method is suitable for periodic analyses in a clinical biochemistry laboratory for endocrinology investigation purposes, simultaneously analysing E and F levels in a saliva specimen.

  18. Gastroprotection and effect of the simultaneous administration of Cuachalalate (Amphipterygium adstringens) on the pharmacokinetics and anti-inflammatory activity of diclofenac in rats.

    Science.gov (United States)

    Navarrete, Andrés; Oliva, Iván; Sánchez-Mendoza, María Elena; Arrieta, Jesús; Cruz-Antonio, Leticia; Castañeda-Hernández, Gilberto

    2005-12-01

    This work aimed to study the effect of Cuachalalate methanol extract (CME) on the anti-inflammatory activity and pharmacokinetics of diclofenac sodium, a frequently prescribed non-steroidal anti-inflammatory drug (NSAID). The gastroprotective effect of CME on the gastric injury induced by diclofenac was studied in rats. CME showed a gastroprotective effect of 15.7% at 1 mg kg(-1) and 72.5% at dose of 300 mg kg(-1). Omeprazole, used as anti-ulcer reference drug, showed gastroprotective effects of 50-89.7% at doses tested (1-30 mg kg(-1)). The value of the 50% effective dose for the anti-inflammatory effect of diclofenac sodium (ED50 = 1.14 +/- 0.23 mg kg(-1)) using carrageenan-induced rat paw oedema model, was not modified by the concomitant administration of 30 or 100 mg kg(-1) of CME. The effect of CME (30, 100 and 300 mg kg(-1), p.o.) on the pharmacokinetics of diclofenac sodium was studied. It was observed that the simultaneous administration of diclofenac sodium and 300 mg kg(-1) of CME decreased significantly the values of Cmax (7.08 +/- 1.42 microg mL(-1)) and AUC (12.67 +/- 2.97 microg h mL(-1)), but not the value of tmax (0.13 (0.1-0.25)h) obtained with the administration of diclofenac alone. The simultaneous administration of 30 or 100 mg kg(-1) of CME did not modify the pharmacokinetic parameters of diclofenac. The experimental findings in rats suggest that CME at doses lower than 100 mg kg(-1) protects the gastric mucosa from the damage induced by diclofenac sodium without altering either the anti-inflammatory activity or the pharmacokinetics of this NSAID.

  19. Simultaneous Determination and Pharmacokinetic Study of Six Components in Rat Plasma by HPLC-MS/MS after Oral Administration of Acanthopanax sessiliflorus Fruit Extract.

    Science.gov (United States)

    Du, Peng; Lei, Mingdao; Liu, Yu; Yang, Shilin

    2016-12-28

    A specific and reliable HPLC-MS/MS method was developed and validated for the simultaneous determination of protocatechuic acid (PCA), scopolin, (-)-pinoresinol-4,4'-di-O-β-d-glucopyranoside (PDG), acanthoside D, acanthoside B and hyperin in rat plasma for the first time. The analytes were separated on a C18 column (50 × 2.1 mm, 1.8 µm) and a triple-quadrupole mass spectrometer equipped with an electrospray ionization (ESI) source was used for detection. The rat plasma sample was prepared using the protein precipitation procedure. The calibration curves were linear over a concentration range of 1.2-1200.0 ng/mL for PCA, 0.96-960.0 ng/mL for scopolin, 1.12-1120.0 ng/mL for PDG, 1.32-1320.0 ng/mL for acanthoside D, 0.99-990.0 ng/mL for acanthoside B and 1.01-1010.0 ng/mL for hyperin. The intra-day and inter-day precision was less than 11.4% and the relative error (RE) was all within ±15%. The validated method was successfully applied to assess the pharmacokinetics characteristics after the extracts of Acanthopanax sessiliflorus fruits were orally administered to the Sprague-Dawley rat.

  20. A sensitive and selective UPLC-MS/MS method for simultaneous determination of 10 alkaloids from Rhizoma Menispermi in rat plasma and its application to a pharmacokinetic study.

    Science.gov (United States)

    Wei, Jinxia; Fang, Linlin; Liang, Xinlei; Su, Dan; Guo, Xingjie

    2015-11-01

    A sensitive and selective liquid chromatography-tandem mass spectrometry method has been developed and validated for simultaneous quantitation of 10 alkaloids (dauricine, daurisoline, N-desmethyldauricine, dauricicoline, dauriporphinoline, bianfugecine, dauricoside, stepholidine, acutumine and acutumidine) from Rhizoma Menispermi in rat plasma. After addition of internal standard (verapamil), plasma samples were pretreated by a single-step protein precipitation with acetonitrile. Chromatographic separation was performed on a Waters BEH C18 column with gradient elution using a mobile phase composed of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.3 mL/min. The analytes were detected without interference in the multiple reaction monitoring (MRM) mode with positive electrospray ionization. The validated method exhibited good linearity over a wide concentration range (r≥0.9914), and the lower limits of quantification were 0.01-5.0 ng/mL for all the analytes. The intra-day and inter-day precisions (RSD) at three different levels were both less than 13.4% and the accuracies (RE) ranged from -12.8% to 13.5%. The mean extraction recoveries of analytes and IS from rat plasma were all more than 77%. The validated method was successfully applied to a comparative pharmacokinetic study of 10 alkaloids in rat plasma after oral administration of Rhizoma Menispermi extract.

  1. Simultaneous determination of catechin, epicatechin and epicatechin gallate in rat plasma by LC-ESI-MS/MS for pharmacokinetic studies after oral administration of Cynomorium songaricum extract.

    Science.gov (United States)

    Zhang, Qiu-Hong; Wang, Wen-Biao; Li, Jin; Chang, Yan-Xu; Wang, Yue-Fei; Zhang, Jishu; Zhang, Bo-Li; Gao, Xiu-Mei

    2012-01-01

    A rapid and valid method was developed for simultaneous determination catechin, epicatechin and epicatechin gallate in rat plasmas using scopoletin (103 ng mL(-1)) as an internal standard (IS). The separation was performed on Eclipse plus C18 column (100 mm × 4.6 mm, 1.8 μm) at a flow rate of 0.3 mL min(-1), and acetonitrile-0.1% formic acid was used as mobile phase. The recoveries of three analytes and IS were more than 78.9%. The lower limits of quantitation (LLOQ) in rat plasma were 2.14, 2.38 and 2.08 ng mL(-1) respectively for catechin, epicatechin and epicatechin gallate. Intra-day and inter-day precisions were within 12%. The accuracies were more than 85%. After single oral administration of 15.25 g kg(-1) Cynomorium songaricum extract, C(max) of catechin, epicatechin and epicatechin gallate in rat plasma were respectively 86.69±38.65, 32.57±15.00 and 36.93±12.62 ng mL(-1) while T(max) values were respectively 0.15±0.09, 0.20±0.10 and 0.20±0.13 h. The results demonstrated that the present LC-MS/MS method was sensitive enough for pharmacokinetic study of catichins following oral administration of C. songaricum extract.

  2. Simultaneous Determination and Pharmacokinetic Study of Six Components in Rat Plasma by HPLC-MS/MS after Oral Administration of Acanthopanax sessiliflorus Fruit Extract

    Directory of Open Access Journals (Sweden)

    Peng Du

    2016-12-01

    Full Text Available A specific and reliable HPLC-MS/MS method was developed and validated for the simultaneous determination of protocatechuic acid (PCA, scopolin, (−-pinoresinol-4,4′-di-O-β-d-glucopyranoside (PDG, acanthoside D, acanthoside B and hyperin in rat plasma for the first time. The analytes were separated on a C18 column (50 × 2.1 mm, 1.8 µm and a triple-quadrupole mass spectrometer equipped with an electrospray ionization (ESI source was used for detection. The rat plasma sample was prepared using the protein precipitation procedure. The calibration curves were linear over a concentration range of 1.2–1200.0 ng/mL for PCA, 0.96–960.0 ng/mL for scopolin, 1.12–1120.0 ng/mL for PDG, 1.32–1320.0 ng/mL for acanthoside D, 0.99–990.0 ng/mL for acanthoside B and 1.01–1010.0 ng/mL for hyperin. The intra-day and inter-day precision was less than 11.4% and the relative error (RE was all within ±15%. The validated method was successfully applied to assess the pharmacokinetics characteristics after the extracts of Acanthopanax sessiliflorus fruits were orally administered to the Sprague-Dawley rat.

  3. Simultaneous Quantification of Glibenclamide, Simvastatin, and Quercetin by Using LC-UV Method and Its Application to Pharmacokinetic Study in Rats

    Directory of Open Access Journals (Sweden)

    Vijay Duppala

    2013-01-01

    Full Text Available A sensitive, precise, and simple LC method for the simultaneous quantification of glibenclamide, simvastatin, and quercetin in rat plasma has been developed and validated. The chromatographic separation was achieved on a cyano column (250 mm × 4.6 mm, 5 µm maintained at room temperature, using isocratic elution with methanol : acetonitrile : 10 mM potassium dihydrogen orthophosphate, pH adjusted to 4.5 with o-phosphoric acid (8 : 32 : 60, v/v and detected using UV-VIS detector. Plasma samples were deproteinated with 0.1% perchloric acid and acetonitrile for extraction of the glibenclamide, simvastatin, and quercetin which resulted in their high recoveries. LC calibration curves based on the extracts from the rat plasma were linear in the range of 50–1000 ng mL−1 for all the three drugs. The limit of quantification was 50 ng mL−1. The described method was successfully applied to study the pharmacokinetics of glibenclamide, simvastatin, and quercetin following oral administration, in combination to Sprague-Dawley rats.

  4. Simultaneous determination of anemoside B4, phellodendrine, berberine, palmatine, obakunone, esculin, esculetin in rat plasma by UPLC-ESI-MS/MS and its application to a comparative pharmacokinetic study in normal and ulcerative colitis rats.

    Science.gov (United States)

    Yang, Lianrong; Meng, Xin; Yu, Xiaojin; Kuang, Haixue

    2017-02-05

    A sensitive and rapid ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was developed for the simultaneous analysis of anemoside B4, phellodendrine, berberine, palmatine, obakunone, esculin, esculetin, toosendanin (IS1 of anemoside B4), tetrahydropalmatine (IS2 of phellodendrine, berberine, palmatine and obakunone) and scopoletin (IS3 of esculin and esculetin) and to compare the pharmacokinetics of these active ingredients in normal and ulcerative colitis rats. After methanol deproteinization, solvents were evaporated at 40°C under a gentle stream of nitrogen. Chromatography was performed using a C18 column with a gradient elution of 0.1% aqueous formic acid and acetonitrile at 0.4ml/min. Detection and measurement were performed on a 4000 QTRAP UPLC-MS/MS system from AB Sciex in the multiple reaction monitoring (MRM) mode. Phellodendrine, berberine, palmatine, obakunone, esculin, esculetin, tetrahydropalmatine (IS2) and scopoletin (IS3) were monitored under positive ionization conditions. Anemoside B4, and toosendanin (IS1) were monitored under negative ionization conditions. The optimized mass transition ion-pairs (m/z) were 1221.1/750.7 for anemoside B4, 343.2/193.2 for phellodendrine, 337.1/321.0 for berberine, 353.0/336.9 for palmatine, 455.1/161.1 for obakunone, 341.2/179.2 for esculin, 179.1/123.0 for esculetin, 573.4/531.4 for toosendanin (IS1), 356.2/192.2 for tetrahydropalmatine (IS2) and 193.0/133.1 for scopoletin (IS3). Copyright © 2016 Elsevier B.V. All rights reserved.

  5. A single extraction and HPLC procedure for simultaneous analysis of phytosterols, tocopherols and lutein in soybeans.

    Science.gov (United States)

    Slavin, Margaret; Yu, Liangli Lucy

    2012-12-15

    A saponification/extraction procedure and high performance liquid chromatography (HPLC) analysis method were developed and validated for simultaneous analysis of phytosterols, tocopherols and lutein (a carotenoid) in soybeans. Separation was achieved on a phenyl column with a ternary, isocratic solvent system of acetonitrile, methanol and water (48:22.5:29.5, v/v/v). Evaporative light scattering detection (ELSD) was used to quantify β-sitosterol, stigmasterol, campesterol, and α-, δ- and γ-tocopherols, while lutein was quantified with visible light absorption at 450 nm. Peak identification was verified by retention times and spikes with external standards. Standard curves were constructed (R(2)>0.99) to allow for sample quantification. Recovery of the saponification and extraction was demonstrated via analysis of spiked samples. Also, the accuracy of results of four soybeans using the described saponification and HPLC analytical method was validated against existing methods. This method offers a more efficient alternative to individual methods for quantifying lutein, tocopherols and sterols in soybeans.

  6. Analysis of Age Dependent Effects of Heat Stress on EEG Frequency Components in Rats

    Institute of Scientific and Technical Information of China (English)

    RAKESH KUMAR SINHA

    2009-01-01

    Objective To demonstrate changes in different frequencies of cerebral electrical activity or electroencephalogram (EEG) following exposure to high environmental heat in three different age groups of freely moving rats. Methods Rats were divided into three groups (i) acute heat stress - subjected to a single exposure for four hours at 38 ℃; (ii) chronic heat stress -exposed for 21 days daily for one hour at 38 ℃, and (iii) handling control groups. The digital polygraphic sleep-EEG recordings were performed just after the heat exposure from acute stressed rats and on 22nd day from chronic stressed rats by simultaneous recording of cortical EEG EOG (electrooculogram), and EMG (electromyogram). Further, power spectrum analyses were performed to analyze the effects of heat stress. Results The frequency analysis of EEG signals following exposure to high environmental heat revealed that in all three age groups of rats, changes in higher frequency components (β2) were significant in all sleep-wake states following both acute and chronic heat stress conditions. After exposure to acute heat, significant changes in EEG frequencies with respect to their control groups were observed, which were reversed partly or fully in four hours of EEG recording. On the other hand, due to repetitive chronic exposure to hot environment, adaptive and long-term changes in EEG frequency patterns were observed. Conclusion The present study has exhibited that the cortical EEG is sensitive to environmental heat and alterations in EEG frequencies in different sleep-wake states due to heat stress can be differentiated efficiently by EEG power spectrum analysis.

  7. A methd for simultaneous measurement of intragastric, intraduodenal, and intracolonic pressures in the bilaterally vagotomized spinal rat

    NARCIS (Netherlands)

    Y.N. Zhu (Y.); K.M. Dhasmana (K.); W. Erdmann (Wilhelm)

    1994-01-01

    textabstractA method has been developed where quantitative evaluation of intragastric (IGP), intraduodenal (IDP), and intracolonic (ICP) pressures (cm H2O) can be obtained in the bilaterally vagotomized spinal rat. The preparation is very sensitive to 5-hydroxytryptamine (5-HT) and 5-HT-agonists, ca

  8. A methd for simultaneous measurement of intragastric, intraduodenal, and intracolonic pressures in the bilaterally vagotomized spinal rat

    NARCIS (Netherlands)

    Y.N. Zhu (Y.); K.M. Dhasmana (K.); W. Erdmann (Wilhelm)

    1994-01-01

    textabstractA method has been developed where quantitative evaluation of intragastric (IGP), intraduodenal (IDP), and intracolonic (ICP) pressures (cm H2O) can be obtained in the bilaterally vagotomized spinal rat. The preparation is very sensitive to 5-hydroxytryptamine (5-HT) and 5-HT-agonists,

  9. Can fusion, elastic and inelastic scattering of heavy ions be understood, without a simultaneous analysis of them?

    Energy Technology Data Exchange (ETDEWEB)

    Anjos, R.M.; Muri, C.; Moraes, S.B.; Cabezas, R.; Gomes, P.R.S.; Tenreiro, C.; Liguori Neto, R.; Maciel, A.M.M.; Santos, G.M. [Instituto de Fisica, Universidade Federal Fluminense, Av. Litoranea s/n, Gragoata, Niteroi, RJ, 24210-340 (Brazil)

    1997-10-01

    We present examples of situations where the absence of a simultaneous analysis of the scattering and reaction mechanisms leads to an incomplete or even false understanding of these processes. The optical model analysis of the elastic scattering gives rise to different values of reaction cross sections, while the simple analysis of the fusion excitation functions may lead to ambiguous or wrong conclusions. Data for the {sup 14}N+{sup 59}Co system obtained by our group is used in the analysis. (author)

  10. Simultaneous determination of seven alkaloids in rat plasma by UFLC-MS/MS and its application to a pharmacokinetic study after oral administration of Cerebralcare Granule.

    Science.gov (United States)

    Xiaowen, Li; Ling, Tong; Yunfei, Li; Guoxiang, Sun; Dailin, Yang; Herry, Sun

    2016-04-01

    An ultra fast liquid chromatography-tandem mass sepectrometry (UFLC-MS/MS) method was developed for simultaneous determination of seven active alkaloid components (tetrahydropalmatine, corydaline, α-allocryptopine, tetrahydroberberine, tetrahydrocoptisine, tetrahydrocolumbamine and dehydrocorydaline) in rat plasma after oral administration of Cerebralcare Granule. Plasma samples were pretreated by protein precipitation with acetronitrile containing the internal standard diazepam. Chromatographic separation was achieved on a Phenomenex Kinetex C18 column (100×2.1mm, 2.6μm) with gradient elution using mobile phase consisting of acetonitrile -0.1% formic acid in water at a flow rate of 0.3mL/min. The detection was performed on an electrospray ionization triple quadrupole tandem mass spectrometer using multiple reaction monitoring (MRM) with positive ionization mode. The established method was fully validated and proved to be sensitive and specific with lower limits of quantification (LLOQs) all less than 0.0265ng/mL in rat plasma. Good linearities of seven alkaloids were obtained in respective concentration ranges (r>0.9923). The intra- and inter-day precisions were below of 15% for all the seven alkaloids in terms of relative standard deviation (RSD), and the accuracies were ranged from -2.7% to 8.3% in terms of relative error (RE). Extraction recovery, matrix effect and stability were within the required limits in rat plasma. The validated method was successfully applied to investigate the pharmacokinetics of the seven alkaloids in rat plasma after oral administration of Cerebralcare Granule (CG).

  11. Development of a simple and sensitive HPLC-UV method for the simultaneous determination of cannabidiol and Δ(9)-tetrahydrocannabinol in rat plasma.

    Science.gov (United States)

    Zgair, Atheer; Wong, Jonathan C M; Sabri, Akmal; Fischer, Peter M; Barrett, David A; Constantinescu, Cris S; Gershkovich, Pavel

    2015-10-10

    There has been increased interest in the medical use of cannabinoids in recent years, particularly in the predominant natural cannabinoids, cannabidiol (CBD) and Δ(9)-tetrahydrocannabinol (THC). The aim of the current study was to develop a sensitive and reliable method for the quantification of CBD and THC in rat plasma. A combination of protein precipitation using cold acetonitrile and liquid-liquid extraction using n-hexane was utilised to extract CBD and THC from rat plasma. Samples were then evaporated and reconstituted in acetonitrile and 30 μL was injected into an HPLC system. Separation was achieved using an ACE C18-PFP 150 mm × 4.6 mm, 3 μm column at 55 °C with isocratic elution using a mobile phase consisting of acetonitrile-water (62:38, v/v) at 1 mL/min for 20 min. Both cannabinoids, as well as the internal standard (4,4-dichlorodiphenyltrichloroethane, DDT) were detected at 220 nm. Our new method showed linearity in the range of 10-10,000 ng/mL and a lower limit of quantification (LLOQ) of 10 ng/mL for both cannabinoids, which is comparable to previously reported LC-MS/MS methods. Inter- and intra-day precision and accuracy were below 15% RSD and RE, respectively. To demonstrate the suitability of the method for in vivo studies in rats, the assay was applied to a preliminary pharmacokinetic study following IV bolus administration of 5 mg/kg CBD or THC. In conclusion, a simple, sensitive, and cost-efficient HPLC-UV method for the simultaneous determination of CBD and THC has been successfully developed, validated and applied to a pharmacokinetic study in rats.

  12. Reevaluation of Stratospheric Ozone Trends From SAGE II Data Using a Simultaneous Temporal and Spatial Analysis

    Science.gov (United States)

    Damadeo, R. P.; Zawodny, J. M.; Thomason, L. W.

    2014-01-01

    This paper details a new method of regression for sparsely sampled data sets for use with time-series analysis, in particular the Stratospheric Aerosol and Gas Experiment (SAGE) II ozone data set. Non-uniform spatial, temporal, and diurnal sampling present in the data set result in biased values for the long-term trend if not accounted for. This new method is performed close to the native resolution of measurements and is a simultaneous temporal and spatial analysis that accounts for potential diurnal ozone variation. Results show biases, introduced by the way data is prepared for use with traditional methods, can be as high as 10%. Derived long-term changes show declines in ozone similar to other studies but very different trends in the presumed recovery period, with differences up to 2% per decade. The regression model allows for a variable turnaround time and reveals a hemispheric asymmetry in derived trends in the middle to upper stratosphere. Similar methodology is also applied to SAGE II aerosol optical depth data to create a new volcanic proxy that covers the SAGE II mission period. Ultimately this technique may be extensible towards the inclusion of multiple data sets without the need for homogenization.

  13. Simultaneous analysis of multiple neurotransmitters by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry.

    Science.gov (United States)

    Tufi, Sara; Lamoree, Marja; de Boer, Jacob; Leonards, Pim

    2015-05-22

    Neurotransmitters are endogenous metabolites that allow the signal transmission across neuronal synapses. Their biological role is crucial for many physiological functions and their levels can be changed by several diseases. Because of their high polarity, hydrophilic interaction liquid chromatography (HILIC) is a promising tool for neurotransmitter analysis. Due to the large number of HILIC stationary phases available, an evaluation of the column performances and retention behaviors has been performed on five different commercial HILIC packing materials (silica, amino, amide and two zwitterionic stationary phases). Several parameters like the linear correlation between retention and the distribution coefficient (logD), the separation factor k and the column resolution Rs have been investigated and the column performances have been visualized with a heat map and hierarchical clustering analysis. An optimized and validated HILIC-MS/MS method based on the ZIC-cHILIC column is proposed for the simultaneous detection and quantification of twenty compounds consisting of neurotransmitters, precursors and metabolites: 3-methoxytyramine (3-MT), 5-hydroxyindoleacetic acid (5-HIAA), 5-hydroxy-L-tripthophan, acetylcholine, choline, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, epinephrine, γ-aminobutyric acid (GABA), glutamate, glutamine, histamine, histidine, L-tryptophan, L-tyrosine, norepinephrine, normetanephrine, phenylalanine, serotonin and tyramine. The method was applied to neuronal metabolite profiling of the central nervous system of the freshwater snail Lymnaea stagnalis. This method is suitable to explore neuronal metabolism and its alteration in different biological matrices.

  14. Simultaneous analysis of consumer variables, acceptability and sensory characteristics of dry-cured ham.

    Science.gov (United States)

    Sugimoto, Masahiro; Obiya, Shinichi; Kaneko, Miku; Enomoto, Ayame; Honma, Mayu; Wakayama, Masataka; Tomita, Masaru

    2016-11-01

    We conducted a consumer acceptability analysis of dry-cured ham based on sensory evaluation. Consumer acceptability data are rendered heterogeneous by the diverse backgrounds and assessment abilities of the participants, requiring versatile analytical methods for their interpretation. Totally, 9 sensory attributes of 12 kinds of dry-cured ham samples collected from Japan (n=9), Italy (n=1), Spain (n=1), and Germany (n=1) were tasted by 117 Japanese consumers who showed acceptable evaluation abilities during blind sampling. Common techniques, such as hierarchical clustering, principal component analysis, and external preference mapping, were simultaneously utilized to analyze each characteristics scored in modified hedonic scale. These analyses revealed the relationships between the features and preferences of the assessors. For example, consumers aged 20-30 with smoking and drinking habits preferred sweetness and saltiness, and gave high ratings to Spanish Jómon serrano and Italian prosciutto. Our approach could assist ham marketers to identify potential purchasers and the preferred characteristics of their products.

  15. Automated simultaneous monitoring of nitrate and nitrite in surface water by sequential injection analysis.

    Science.gov (United States)

    Legnerová, Zlatuse; Solich, Petr; Sklenárová, Hana; Satínský, Dalibor; Karlícek, Rolf

    2002-06-01

    A fully automated procedure based on Sequential Injection Analysis (SIA) methodology for simultaneous monitoring of nitrate and nitrite in surface water samples is described. Nitrite was determined directly using the Griess diazo-coupling reaction and the formed azo dye was measured at 540 nm in the flow cell of the fibre-optic spectrophotometer. Nitrate zone was passed through a reducing mini-column containing copperised-cadmium. After the reduction of nitrate into nitrite the sample was aspirated by flow reversal to the holding coil, treated with the reagent and finally passed through the flow cell. The calibration curve was linear over the range 0.05-1.00 mg N l(-1) of nitrite and 0.50-50.00 mg N l(-1) of nitrate; correlation coefficients were 0.9993 and 0.9988 for nitrite and nitrate, respectively. Detection limits were 0.015 and 0.10 mg N l(-1) for nitrite and nitrate, respectively. The relative standard deviation (RSD) values (n = 3) were 1.10% and 1.32% for nitrite and nitrate, respectively. The total time of one measuring cycle was 250 s, thus the sample throughput was about 14 h(-1). Nitrate and nitrite were determined in the real samples of surface water, and the results have been compared with those obtained by two other flow methods; flow injection analysis based on the same reactions and isotachophoretic determination used in a routine environmental control laboratory.

  16. Simultaneous analysis of noble gases, sulfur hexafluoride, and other dissolved gases in water.

    Science.gov (United States)

    Brennwald, Matthias S; Hofer, Markus; Kipfer, Rolf

    2013-08-06

    We developed an analytical method for the simultaneous measurement of dissolved He, Ne, Ar, Kr, Xe, SF6, N2, and O2 concentrations in a single water sample. The gases are extracted from the water using a head space technique and are transferred into a vacuum system for purification and separation into different fractions using a series of cold traps. Helium is analyzed using a quadrupole mass spectrometer (QMS). The remaining gas species are analyzed using a gas chromatograph equipped with a mass spectrometer (GC-MS) for analysis of Ne, Ar, Kr, Xe, N2, and O2 and an electron capture detector (GC-ECD) for SF6 analysis. Standard errors of the gas concentrations are approximately 8% for He and 2-5% for the remaining gas species. The method can be extended to also measure concentrations of chlorofluorocarbons (CFCs). Tests of the method in Lake Lucerne (Switzerland) showed that dissolved gas concentrations agree with measurements from other methods and concentrations of air saturated water. In a small artificial pond, we observed systematic gas supersaturations, which seem to be linked to adsorption of solar irradiation in the pond and to water circulation through a gravel bed.

  17. Cordyceps sinensis protects against liver and heart injuries in a rat model of chronic kidney disease: a metabolomic analysis

    Science.gov (United States)

    Liu, Xia; Zhong, Fang; Tang, Xu-long; Lian, Fu-lin; Zhou, Qiao; Guo, Shan-mai; Liu, Jia-fu; Sun, Peng; Hao, Xu; Lu, Ying; Wang, Wei-ming; Chen, Nan; Zhang, Nai-xia

    2014-01-01

    Aim: To test the hypothesis that the traditional Chinese medicine Cordyceps sinensis could improve the metabolic function of extrarenal organs to achieve its anti-chronic kidney disease (CKD) effects. Methods: Male SD rats were divided into CKD rats (with 5/6-nephrectomy), CKD rats treated with Cordyceps sinensis (4 mg•kg-1•d-1, po), and sham-operated rats. After an 8-week treatment, metabolites were extracted from the hearts and livers of the rats, and then subjected to 1H-NMR-based metabolomic analysis. Results: Oxidative stress, energy metabolism, amino acid and protein metabolism and choline metabolism were considered as links between CKD and extrarenal organ dysfunction. Within the experimental period of 8 weeks, the metabolic disorders in the liver were more pronounced than in the heart, suggesting that CKD-related extrarenal organ dysfunctions occurred sequentially rather than simultaneously. Oral administration of Cordyceps sinensis exerted statistically significant rescue effects on the liver and heart by reversely regulating levels of those metabolites that are typically perturbed in CKD. Conclusion: Oral administration of Cordyceps sinensis significantly attenuates the liver and heart injuries in CKD rats. The 1H NMR-based metabolomic approach has provided a systematic view for understanding of CKD and the drug treatment, which can also be used to elucidate the mechanisms of action of other traditional Chinese medicines. PMID:24632844

  18. Simultaneous and independent optical impairments monitoring using singular spectrum analysis of asynchronously sampled signal amplitudes

    Science.gov (United States)

    Guesmi, Latifa; Menif, Mourad

    2015-09-01

    Optical performance monitoring (OPM) becomes an inviting topic in high speed optical communication networks. In this paper, a novel technique of OPM based on a new elaborated computation approach of singular spectrum analysis (SSA) for time series prediction is presented. Indeed, various optical impairments among chromatic dispersion (CD), polarization mode dispersion (PMD) and amplified spontaneous emission (ASE) noise are a major factors limiting quality of transmission data in the systems with data rates lager than 40 Gbit/s. This technique proposed an independent and simultaneous multi-impairments monitoring, where we used SSA of time series analysis and forecasting. It has proven their usefulness in the temporal analysis of short and noisy time series in several fields, that it is based on the singular value decomposition (SVD). Also, advanced optical modulation formats (100 Gbit/s non-return-to zero dual-polarization quadrature phase shift keying (NRZ-DP-QPSK) and 160 Gbit/s DP-16 quadrature amplitude modulation (DP-16QAM)) offering high spectral efficiencies have been successfully employed by analyzing their asynchronously sampled amplitude. The simulated results proved that our method is efficient on CD, first-order PMD, Q-factor and OSNR monitoring, which enabled large monitoring ranges, the CD in the range of 170-1700 ps/nm.Km and 170-1110 ps/nm.Km for 100 Gbit/s NRZ-DP-QPSK and 160 Gbit/s DP-16QAM respectively, and also the DGD up to 20 ps is monitored. We could accurately monitor the OSNR in the range of 10-40 dB with monitoring error remains less than 1 dB in the presence of large accumulated CD.

  19. Separate urinary bladder and prostate neurons in the central nervous system of the rat: simultaneous labeling with two immunohistochemically distinguishable pseudorabies viruses

    Directory of Open Access Journals (Sweden)

    Nadelhaft Irving

    2002-07-01

    Full Text Available Abstract Background This work examines the central nervous system distribution of virus-labeled neurons from the rat urinary bladder and the prostate simultaneously within the same tissue sections. Two immunohistochemically distinct pseudorabies virus strains were simultaneously injected into male Sprague Dawley rats (~280 gm. One virus was injected into the bladder and the other into the prostate. After incubation intervals of 2.25, 2.5, 2.75, 3 and 4 days, sections from the spinal cord and brain were processed immunohistochemically to detect cells, within a single section, which were labeled separately by each virus or were labeled by both viruses. Results Each strain of virus labeled a separate population of neurons and some neurons were labeled by both strains. The majority of neurons labeled by virus from the urinary bladder were found in the L6-S1 spinal cord segments within the dorsal gray commissure, the intermediolateral area and the superficial dorsal horn. Neurons labeled by virus from the prostate were mainly found in the L1-L2 spinal cord segments in the dorsal gray commissure and the intermediolateral areas. Double-labeled interneurons in L1-L2 were mainly located in the intermediolateral area. In L6-S1 they were divided between the dorsal gray commissure and the intermediolateral area. Conclusions Spinal neurons innervating the bladder are clearly separate and different from those innervating the prostate. This difference also persists in the brain. In disagreement with previous reports, no direct anatomical evidence of parasympathetic innervation of the prostate was observed.

  20. Simultaneous determination of four phenolic acids and seven alkaloids in rat plasma after oral administration of traditional Chinese medicinal preparation Jinqi Jiangtang Tablet by LC-ESI-MS/MS.

    Science.gov (United States)

    Chang, Yan-xu; Ge, Ai-hua; Yu, Xie-an; Jiao, Xiu-cheng; Li, Jin; He, Jun; Tian, Ji; Liu, Wei; Azietaku, John Teye; Zhang, Bo-li; Gao, Xiu-mei

    2016-01-05

    A rapid, sensitive and selective high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of four phenolic acids (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and ferulic acid) and seven alkaloids (berberine, epiberberine, coptisine, magnoflorine, berberubine, palmatine and jatrorrhizine) in rat plasma. After mixing with the internal standards tetrahydropalmatine (IS1) and rosmarinic acid (IS2), plasma samples were pretreated by protein precipitation using acetonitrile. The HPLC analysis was performed on an Agilent Eclipse plus C18 (4.6 mm×100 mm, 1.8 μm) column with mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile at a flow rate of 0.3 mL min(-1). The detection was accomplished for the analytes and internal standards using positive electrospray ionization for the alkaloids and negative electrospray ionization for the phenolic acids in multiple-reaction monitoring mode. The method showed a good linearity over a wide concentration range (r(2)>0.99). The lower limit of quantification of seven alkaloids was lower than 2 ng mL(-1) and that of four phenolic acids was less than 20 ng mL(-1). The developed method was applied to the pharmacokinetic study of 11 components after oral administration of traditional Chinese medicinal preparation Jinqi Jiangtang Tablet in rats.

  1. Simultaneous analysis of synthetic cannabinoids in the materials seized during drug trafficking using GC-MS.

    Science.gov (United States)

    Choi, Hyeyoung; Heo, Sewoong; Choe, Sanggil; Yang, Wonkyung; Park, Yuran; Kim, Eunmi; Chung, Heesun; Lee, Jaesin

    2013-05-01

    A rapid and simple gas chromatography-mass spectrometry (GC-MS) method was developed and validated to identify and quantify synthetic cannabinoids in the materials seized during drug trafficking. Accuracy and reproducibility of the method were improved by using deuterated JWH-018 and JWH-073 as internal standards. Validation results of the GC-MS method showed that it was suitable for simultaneous qualitative and quantitative analyses of synthetic cannabinoids, and we analyzed synthetic cannabinoids in seized materials using the validated GC-MS method. As a result of the analysis, ten species of synthetic cannabinoids were identified in dried leaves (n = 40), bulk powders (n = 6), and tablets (n = 14) seized in Korea during 2009-2012, as a single ingredient or as a mixture with other active co-ingredients. JWH-018 and JWH-073 were the most frequently identified compounds in the seized materials. Synthetic cannabinoids in the dried leaves showed broad concentration ranges, which may cause unexpected toxicity to abusers. The bulk powders were considered as raw materials used to prepare legal highs, and they contained single ingredient of JWH-073, JWH-019, or JWH-250 with the purity over 70 %. In contrast, JWH-018 and JWH-073 contents in the tablets were 7.1-13.8 and 3.0-10.2 mg/g, respectively. Relatively low contents in the tablets suggest that the synthetic cannabinoids may have been added to the tablets as supplements to other active co-ingredients.

  2. Gender, space, and the location changes of jobs and people: a spatial simultaneous equations analysis.

    Science.gov (United States)

    Hoogstra, Gerke J

    2012-01-01

    This article summarizes a spatial econometric analysis of local population and employment growth in the Netherlands, with specific reference to impacts of gender and space. The simultaneous equations model used distinguishes between population- and gender-specific employment groups, and includes autoregressive and cross-regressive spatial lags to detect relations both within and among these groups. Spatial weights matrices reflecting different bands of travel times are used to calculate the spatial lags and to gauge the spatial nature of these relations. The empirical results show that although population–employment interaction is more localized for women's employment, no gender difference exists in the direction of interaction. Employment growth for both men and women is more influenced by population growth than vice versa. The interaction within employment groups is even more important than population growth. Women's, and especially men's, local employment growth mostly benefits from the same employment growth in neighboring locations. Finally, interaction between these groups is practically absent, although men's employment growth may have a negative impact on women's employment growth within small geographic areas. In summary, the results confirm the crucial roles of gender and space, and offer important insights into possible relations within and among subgroups of jobs and people.

  3. Simultaneous quantitative analysis of main components in linderae reflexae radix with one single marker.

    Science.gov (United States)

    Wang, Li-Li; Zhang, Yun-Bin; Sun, Xiao-Ya; Chen, Sui-Qing

    2016-05-08

    Establish a quantitative analysis of multi-components by the single marker (QAMS) method for quality evaluation and validate its feasibilities by the simultaneous quantitative assay of four main components in Linderae Reflexae Radix. Four main components of pinostrobin, pinosylvin, pinocembrin, and 3,5-dihydroxy-2-(1-p-mentheneyl)-trans-stilbene were selected as analytes to evaluate the quality by RP-HPLC coupled with a UV-detector. The method was evaluated by a comparison of the quantitative results between the external standard method and QAMS with a different HPLC system. The results showed that no significant differences were found in the quantitative results of the four contents of Linderae Reflexae Radix determined by the external standard method and QAMS (RSD <3%). The contents of four analytes (pinosylvin, pinocembrin, pinostrobin, and Reflexanbene I) in Linderae Reflexae Radix were determined by the single marker of pinosylvin. This fingerprint was the spectra determined by Shimadzu LC-20AT and Waters e2695 HPLC that were equipped with three different columns.

  4. Simultaneous Gaussian and exponential inversion for improved analysis of shales by NMR relaxometry

    Science.gov (United States)

    Washburn, Kathryn E.; Anderssen, Endre; Vogt, Sarah J.; Seymour, Joseph D.; Birdwell, Justin E.; Kirkland, Catherine M.; Codd, Sarah L.

    2014-01-01

    Nuclear magnetic resonance (NMR) relaxometry is commonly used to provide lithology-independent porosity and pore-size estimates for petroleum resource evaluation based on fluid-phase signals. However in shales, substantial hydrogen content is associated with solid and fluid signals and both may be detected. Depending on the motional regime, the signal from the solids may be best described using either exponential or Gaussian decay functions. When the inverse Laplace transform, the standard method for analysis of NMR relaxometry results, is applied to data containing Gaussian decays, this can lead to physically unrealistic responses such as signal or porosity overcall and relaxation times that are too short to be determined using the applied instrument settings. We apply a new simultaneous Gaussian-Exponential (SGE) inversion method to simulated data and measured results obtained on a variety of oil shale samples. The SGE inversion produces more physically realistic results than the inverse Laplace transform and displays more consistent relaxation behavior at high magnetic field strengths. Residuals for the SGE inversion are consistently lower than for the inverse Laplace method and signal overcall at short T2 times is mitigated. Beyond geological samples, the method can also be applied in other fields where the sample relaxation consists of both Gaussian and exponential decays, for example in material, medical and food sciences.

  5. Successive determination of urinary bilirubin and creatinine employing simultaneous injection effective mixing flow analysis.

    Science.gov (United States)

    Ponhong, Kraingkrai; Teshima, Norio; Grudpan, Kate; Vichapong, Jitlada; Motomizu, Shoji; Sakai, Tadao

    2015-02-01

    A novel four-channel simultaneous injection effective mixing flow analysis (SIEMA) system has been assembled for successive determination of bilirubin and creatinine in urinary samples. The chemical variables and physical parameters in the flow system were optimized for the enhancement of successive analytical performances. The interferences from urine matrices on the determination of bilirubin and creatinine were eliminated to dilute urine samples. The calibration graphs with the optimum conditions were achieved to be in 0.024-5.0 mg L(-1) for bilirubin and 2-100 mg L(-1) for creatinine. The relative standard deviations (RSDs) at 3 mg L(-1) of bilirubin and at 50 mg L(-1) of creatinine for 11 runs were 1.5 and 1.0%, respectively. The limits of detections (3σ of blank) for bilirubin and creatinine were 7 µg L(-1) and 0.6 mg L(-1), respectively. The sample throughput for stepwise detection was 22 h(-1). The proposed method was applied to the successive determination of bilirubin and creatinine in urine samples.

  6. Simultaneous preparation of RNA and nuclei for Northern blot and flow cytometric analysis

    Energy Technology Data Exchange (ETDEWEB)

    Tesfaigzi, J.; Jaramillo, R. [Inhalation Toxicology Research Inst., Albuquerque, NM (United States)

    1996-11-01

    Several methods have been developed to quantify RNA synthesis during the progression of the cell cycle. The rate of RNA synthesis can be detected during different stages of the cell cycle by staining cells with agents that intercalate with nucleic acids. For example, following staining of mammalian cells with acridin orange, the green and red fluorescence that correlates with DNA and RNA content, respectively, can be analyzed by flow cytometry. Increase in RNA content during the progression of cells through the cell cycle can be measured after staining with acridin orange. RNA synthesis resulting from the stimulation of quiescent cells with various growth factors has also been demonstrated by labeling cells with bromo-uridine and using the anti-bromo-deoxyuridine antibody. These methods allow measurement of the overall RNA content in cells; however, they do not allow the measurement of the levels of specific mRNAs throughout the cell cycle. Current methods to quantify specific mRNAs generally require the preparation of a large number of cells (5--10 {times} 10{sup 6} cells) to carry out flow cytometric analyses and to isolate RNA for Northern blot analysis or solution hybridization. In this report, the authors describe a method of simultaneously preparing RNA and nuclei for Northern blot and flow cytometric analyses, respectively. The minimum number of nuclei required to obtain flow cytometric data and the effect of conserving nuclei in methanol for several days are also presented.

  7. Performance Analysis of the Microsoft Kinect Sensor for 2D Simultaneous Localization and Mapping (SLAM Techniques

    Directory of Open Access Journals (Sweden)

    Kamarulzaman Kamarudin

    2014-12-01

    Full Text Available This paper presents a performance analysis of two open-source, laser scanner-based Simultaneous Localization and Mapping (SLAM techniques (i.e., Gmapping and Hector SLAM using a Microsoft Kinect to replace the laser sensor. Furthermore, the paper proposes a new system integration approach whereby a Linux virtual machine is used to run the open source SLAM algorithms. The experiments were conducted in two different environments; a small room with no features and a typical office corridor with desks and chairs. Using the data logged from real-time experiments, each SLAM technique was simulated and tested with different parameter settings. The results show that the system is able to achieve real time SLAM operation. The system implementation offers a simple and reliable way to compare the performance of Windows-based SLAM algorithm with the algorithms typically implemented in a Robot Operating System (ROS. The results also indicate that certain modifications to the default laser scanner-based parameters are able to improve the map accuracy. However, the limited field of view and range of Kinect’s depth sensor often causes the map to be inaccurate, especially in featureless areas, therefore the Kinect sensor is not a direct replacement for a laser scanner, but rather offers a feasible alternative for 2D SLAM tasks.

  8. Elemental composition method for computation and analysis of simultaneous chemical and phase equilibrium

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    An elemental composition method for computation and analysis of simultaneous chemical and phase equilibrium (CPE) of non-ideal mixtures is proposed. The concept of element is defined, the relationship between component composition and elemental composition is derived, and the concept of elemental potential and its physical meaning are further cleared from the view point of thermodynamics. The relationship between chemical potential and elemental potential is derived in the thermodynamic principles, and the computation equations for CPE problem are obtained based on elemental potential. A simple form of necessary and sufficient condition in terms of elemental composition for reactive azeotropes is derived, which takes the same functional form as the condition for azeotropes in non-reactive systems. The element in this note may be atoms, molecules or group radicals. The presented method is applicable to CPE problem of non-ideal mixtures, and the computation can be simplified by the dimension reducing method. The presented method was supplied to compute and analyze CPE problem of several examples and it is found that it is a robust and efficient method.

  9. Simultaneous Analysis of Secondary Structure and Light Scattering from Circular Dichroism Titrations: Application to Vectofusin-1

    Science.gov (United States)

    Vermeer, Louic S.; Marquette, Arnaud; Schoup, Michel; Fenard, David; Galy, Anne; Bechinger, Burkhard

    2016-12-01

    Circular Dichroism data are often decomposed into their constituent spectra to quantify the secondary structure of peptides or proteins but the estimation of the secondary structure content fails when light scattering leads to spectral distortion. If peptide-induced liposome self-association occurs, subtracting control curves cannot correct for this. We show that if the cause of the light scattering is independent from the peptide structural changes, the CD spectra can be corrected using principal component analysis (PCA). The light scattering itself is analysed and found to be in good agreement with backscattering experiments. This method therefore allows to simultaneously follow structural changes related to peptide-liposome binding as well as peptide induced liposome self-association. We apply this method to study the structural changes and liposome binding of vectofusin-1, a transduction enhancing peptide used in lentivirus based gene therapy. Vectofusin-1 binds to POPC/POPS liposomes, causing a reversal of the negative liposome charge at high peptide concentrations. When the peptide charges exactly neutralise the lipid charges on both leaflets reversible liposome self-association occurs. These results are in good agreement with biological observations and provide further insight into the conditions required for efficent transduction enhancement.

  10. Performance analysis of the Microsoft Kinect sensor for 2D Simultaneous Localization and Mapping (SLAM) techniques.

    Science.gov (United States)

    Kamarudin, Kamarulzaman; Mamduh, Syed Muhammad; Shakaff, Ali Yeon Md; Zakaria, Ammar

    2014-12-05

    This paper presents a performance analysis of two open-source, laser scanner-based Simultaneous Localization and Mapping (SLAM) techniques (i.e., Gmapping and Hector SLAM) using a Microsoft Kinect to replace the laser sensor. Furthermore, the paper proposes a new system integration approach whereby a Linux virtual machine is used to run the open source SLAM algorithms. The experiments were conducted in two different environments; a small room with no features and a typical office corridor with desks and chairs. Using the data logged from real-time experiments, each SLAM technique was simulated and tested with different parameter settings. The results show that the system is able to achieve real time SLAM operation. The system implementation offers a simple and reliable way to compare the performance of Windows-based SLAM algorithm with the algorithms typically implemented in a Robot Operating System (ROS). The results also indicate that certain modifications to the default laser scanner-based parameters are able to improve the map accuracy. However, the limited field of view and range of Kinect's depth sensor often causes the map to be inaccurate, especially in featureless areas, therefore the Kinect sensor is not a direct replacement for a laser scanner, but rather offers a feasible alternative for 2D SLAM tasks.

  11. Simultaneous Analysis of Secondary Structure and Light Scattering from Circular Dichroism Titrations: Application to Vectofusin-1

    Science.gov (United States)

    Vermeer, Louic S.; Marquette, Arnaud; Schoup, Michel; Fenard, David; Galy, Anne; Bechinger, Burkhard

    2016-01-01

    Circular Dichroism data are often decomposed into their constituent spectra to quantify the secondary structure of peptides or proteins but the estimation of the secondary structure content fails when light scattering leads to spectral distortion. If peptide-induced liposome self-association occurs, subtracting control curves cannot correct for this. We show that if the cause of the light scattering is independent from the peptide structural changes, the CD spectra can be corrected using principal component analysis (PCA). The light scattering itself is analysed and found to be in good agreement with backscattering experiments. This method therefore allows to simultaneously follow structural changes related to peptide-liposome binding as well as peptide induced liposome self-association. We apply this method to study the structural changes and liposome binding of vectofusin-1, a transduction enhancing peptide used in lentivirus based gene therapy. Vectofusin-1 binds to POPC/POPS liposomes, causing a reversal of the negative liposome charge at high peptide concentrations. When the peptide charges exactly neutralise the lipid charges on both leaflets reversible liposome self-association occurs. These results are in good agreement with biological observations and provide further insight into the conditions required for efficent transduction enhancement. PMID:28004740

  12. A new mapping acquisition and processing system for simultaneous PIXE-RBS analysis with external beam

    Science.gov (United States)

    Pichon, L.; Beck, L.; Walter, Ph.; Moignard, B.; Guillou, T.

    2010-06-01

    The combination of ion beam analysis techniques is particularly fruitful for the study of cultural heritage objects. For several years, the AGLAE facility of the Louvre laboratory has been implementing these techniques with an external beam. The recent set-up permits to carry out PIXE, PIGE and RBS simultaneously on the same analyzed spot with a particle beam of approximately 20 μm diameter. A new mapping system has been developed in order to provide elemental concentration maps from the PIXE and RBS spectra. This system combines the Genie2000 spectroscopy software with a homemade software that creates maps by handling acquisition with the object position. Each pixel of each PIXE and RBS maps contains the spectrum normalised by the dose. After analysing each pixel of the PIXE maps (low and high energy X-ray spectra) with the Gupixwin peak-fitting software, quantitative elemental concentrations are obtained for the major and trace elements. This paper presents the quantitative elemental maps extracted from the PIXE spectra and the development of RBS data processing for light element distribution and thin layer characterization. Examples on rock painting and lustrous ceramics will be presented.

  13. In situ simultaneous analysis of polyacetylenes, carotenoids and polysaccharides in carrot roots.

    Science.gov (United States)

    Baranska, Malgorzata; Schulz, Hartwig; Baranski, Rafal; Nothnagel, Thomas; Christensen, Lars P

    2005-08-24

    This paper presents an approach to simultaneously analyze polyacetylenes, carotenoids, and polysaccharides in carrot (Daucus carota L.) roots by means of Raman spectroscopy. The components were measured in situ in the plant tissue without any preliminary sample preparation. The analysis is based on the intensive and characteristic key bands observed in the Raman spectrum of carrot root. The molecular structures of the main carrot polyacetylenes, falcarinol and falcarindiol, are similar, but their Raman spectra exhibit specific differences demonstrated by the shift of their -C[triple bond]C- mode from 2258 to 2252 cm(-)(1), respectively. Carotenoids can be identified by -C=C- stretching vibrations (about 1520 and 1155 cm(-)(1)) of the conjugated system of their polyene chain, whereas the characteristic Raman band at 478 cm(-)(1) indicates the skeletal vibration mode of starch molecule. The other polysaccharide, pectin, can be identified by the characteristic band at 854 cm(-)(1), which is due to the -C-O-C- skeletal mode of alpha-anomer carbohydrates. The Raman mapping technique applied here has revealed detailed information regarding the relative distribution of polyacetylenes, carotenoids, starch, and pectin in the investigated plant tissues. The distribution of these components varies among various carrot cultivars, and especially a significant difference can be seen between cultivated carrot and the wild relative D. carota ssp. maritimus.

  14. Simultaneous Measurement and Analysis of Noise Levels in Flexible and Rigid Pavements

    Directory of Open Access Journals (Sweden)

    Hakan Yıldırım

    2014-06-01

    Full Text Available Although concrete roads have been used worldwide for years, the same improvement could not exist in Turkey and therefore, bituminous (hot mix asphalt roads were preferred instead.In this paper, the vastly built HMA (Hot Mix Asphalt roads and rarely preferred concrete roads were compared based on their level of noise. For this purpose, the concrete road at the length of 2 kilometers between Afyonkarahisar and Emirdag; also the transition point to the HMA road (which continues after the concrete road was observed.Both concrete and HMA road ends of this transition point was equipped with a noise measurement device and a camera was installed separately for providing minimum level of noise interference. Consequently, change in the noise levels depending on the building material of roads was recorded simultaneously. These factors was analysed and various models related to the sort of coating was provided. At this stage, the distance between set up points was kept as short as possible in order to prevent different results in the density of traffic and also in the flow rate.Result of various measurements and analysis provided the noise levels of concrete roads being 4 dB(A less than HMA roads in the comparison based on the same level of vehicle composition and traffic flow. Encouraging the construction of concrete roads in our country is emphasized accordingly.

  15. Simultaneous immunoassay analysis of plasma IL-6 and TNF-α on a microchip.

    Directory of Open Access Journals (Sweden)

    Kaori Abe

    Full Text Available Sandwich enzyme-linked immunosorbant assay (ELISA using a 96-well plate is frequently employed for clinical diagnosis, but is time-and sample-consuming. To overcome these drawbacks, we performed a sandwich ELISA on a microchip. The microchip was made of cyclic olefin copolymer with 4 straight microchannels. For the construction of the sandwich ELISA for interleukin-6 (IL-6 or tumor necrosis factor-α (TNF-α, we used a piezoelectric inkjet printing system for the deposition and fixation of the 1st anti-IL-6 antibody or 1st anti-TNF-α antibody on the surface of the each microchannel. After the infusion of 2 µl of sample to the microchannel and a 20 min incubation, 2 µl of biotinylated 2nd antibody for either antigen was infused and a 10 min incubation. Then 2 µl of avidin-horseradish peroxidase was infused; and after a 5 min incubation, the substrate for peroxidase was infused, and the luminescence intensity was measured. Calibration curves were obtained between the concentration and luminescence intensity over the range of 0 to 32 pg/ml (IL-6: R(2 = 0.9994, TNF-α: R(2 = 0.9977, and the detection limit for each protein was 0.28 pg/ml and 0.46 pg/ml, respectively. Blood IL-6 and TNF-α concentrations of 5 subjects estimated from the microchip data were compared with results obtained by the conventional method, good correlations were observed between the methods according to linear regression analysis (IL-6: R(2 = 0.9954, TNF-α: R(2 = 0.9928. The reproducibility of the presented assay for the determination of the blood IL-6 and TNF-α concentration was comparable to that obtained with the 96-well plate. Simultaneous detection of blood IL-6 and TNF-α was possible by the deposition and fixation of each 1st antibody on the surface of a separate microchannel. This assay enabled us to determine simultaneously blood IL-6 and TNF-α with accuracy, satisfactory sensitivity, time saving ability, and low consumption of sample and

  16. Data supporting the rat brain sample preparation and validation assays for simultaneous determination of 8 neurotransmitters and their metabolites using liquid chromatography–tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Aneta Wojnicz

    2016-06-01

    Full Text Available The data presented in this article supports the rat brain sample preparation procedure previous to its injection into the liquid chromatography–tandem mass spectrometry (LC–MS/MS system to monitor levels of adrenaline, noradrenaline, glutamic acid, γ-aminobutyric acid, dopamine, 5-hydroxytryptamine, 5-hydroxyindole acetic acid, and 3-methoxy-4-hydroxyphenylglycol. In addition, we describe the method validation assays (such as calibration curve, lower limit of quantification, precision and accuracy intra- and inter-day, selectivity, extraction recovery and matrix effect, stability, and carry-over effect according to the United States Food and Drug Administration and European Medicine Agency to measure in one step different neurotransmitters and their metabolites. The data supplied in this article is related to the research study entitled: “Simultaneous determination of 8 neurotransmitters and their metabolite levels in rat brain using liquid chromatography in tandem with mass spectrometry: application to the murine Nrf2 model of depression” (Wojnicz et al. 2016 [1].

  17. Simultaneous determination of two epimeric furofuran lignans (sesamin and asarinin) of Asarum heterotropoides extract in rat plasma by LC/MS/MS: application to pharmacokinetic study.

    Science.gov (United States)

    Ma, Yingyan; Xu, Kai; Wang, Shumin; Han, Yaling

    2014-09-01

    A rapid, sensitive and selective liquid chromatography-tandem mass spectrometry was developed to determine two epimeric furofuran lignans (sesamin and asarinin) simultaneously from Asarum heterotropoides extract in rat plasma. Simple protein precipitation with acetonitrile was performed to extract analytes by using alantolactone as an internal standard. Chromatographic separation was achieved using a DIKMA Diamonsil C18 analytical column (4.6 mm × 150 mm, i.d., 5 µm) by isocratically eluting with a mobile phase consisting of methanol/5 mM ammonium acetate/formic acid (75:25:0.1, v/v/v) at a flow rate of 0.8 mL/min. Tandem mass spectrometric detection with an electrospray ionization interface was performed by multiple reaction monitoring in positive ionization mode. This method was validated according to specificity, sensitivity, linearity, intra- and inter-day precision (sesamin and 5.00-3 000 ng/mL for asarinin. This method has been successfully applied in a pharmacokinetic study of A. heterotropoides extract containing sesamin and asarinin after this extract was orally administrated in rats.

  18. Simultaneous quantification of three alkaloids of Coptidis Rhizoma in rat urine by high-performance liquid chromatography: application to pharmacokinetic study.

    Science.gov (United States)

    Tan, Bo; Ma, Yueming; Shi, Rong; Wang, Tianming

    2007-12-01

    A high-performance liquid chromatographic method with ultraviolet detection was established and validated for quantification of three alkaloids (coptisine, palmatine and berberine) in rat urine. Following a single-step liquid-liquid extraction, the analytes were separated on a reversed-phase C(18) column with water-formic acid-triethylamine-methanol as the mobile phase at a flow rate of 1 ml/min. The linear ranges of the calibration curves were 1.6-160 ng/ml for all three alkaloids. The lower limit of quantification was 1.6 ng/ml for all three alkaloids. The within-batch accuracy was 90.4-108.3% for coptisine, 88.6-107.8% for berberine and 88.4-110.1% for palmatine. The between-batch accuracy was 99.3-100.3% for coptisine, 94.3-100.6% for berberine and 93.7-100.0% for palmatine. The within-batch and between-batch precisions were alkaloids of Coptidis Rhizoma in urine simultaneously and thus to investigate the pharmacokinetics of the alkaloids from Xiexin Decoction in rats.

  19. Simultaneous determination of four Sudan dyes in rat blood by UFLC-MS/MS and its application to a pharmacokinetic study in rats☆

    Institute of Scientific and Technical Information of China (English)

    Hao Zhu; Yijun Chen; Changshun Huanga; Yangyang Han; Yiwei Zhang; Shucan Xie; Xiaohong Chen; Micong Jin

    2015-01-01

    A rapid and sensitive method based on ultrafast liquid chromatography-tandem mass spectrometry was developed and validated for simultaneous determination of Sudan Ⅰ, Sudan Ⅱ, Sudan Ⅲ, and Sudan Ⅳ levels in rat whole blood. Cleanert C18 mixed-mode polymeric sorbent was used for effective solid-phase extraction cleanup. Separation was carried out on a reversed-phase C18 column (100 mm × 2.1 mm, 1.8 μm) using 0.1% (v/v) formic acid in water/0.1% (v/v) formic acid in acetonitrile as the mobile phase in gradient elution. Quantification was performed by an electrospray ionization source in the positive multiple reaction monitoring mode using D5-Sudan I as the internal standard. Calibration curves showed good linearity between 0.2 and 20.0 μg/L, with correlation coefficients higher than 0.9990. The average recovery rates were between 93.05% and 114.98%. The intra- and inter-day relative standard deviations were within 6.2%. The lower limit of quantification was 0.2 μg/L. All the analytes were found to be stable in aseries of stability studies. The proposed method was successfully applied to a pharmacokinetic study of four Sudan dyes after oral administration to rats.

  20. Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler's murine encephalomyelitis virus and rat theilovirus.

    Science.gov (United States)

    Yuan, Wen; Wang, Jing; Xu, Fengjiao; Huang, Bihong; Lian, Yuexiao; Rao, Dan; Yin, Xueqin; Wu, Miaoli; Zhu, Yujun; Zhang, Yu; Huang, Ren; Guo, Pengju

    2016-10-01

    Theiler's murine encephalomyelitis virus (TMEV) and rat theilovirus (RTV), the member of the genus Cardiovirus, are widespread in laboratory mice and rats, and are potential contaminants of biological materials. Cardioviruses infection may cause serious complications in biomedical research. To improve the efficiency of routine screening for Cardioviruses infection, a duplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for simultaneous detection and differentiation of TMEV and RTV. The duplex assay was specific for reference strains of TMEV and RTV, and no cross-reaction was found with seven other rodent viruses. The limits of detection of both TMEV and RTV were 4×10(1) copies RNA/reaction. Reproducibility was estimated using standard dilutions, with coefficients of variation PCR and conventional RT-PCR. For 439 clinical samples,95 samples were positive for TMEV and 72 samples were positive for RTV using duplex real-time RT-PCR approach, whereas only 77 samples were positive for TMEV and 66 samples were positive for RTV when conventional RT-PCR was applied. Mixed infections were found in 20 samples when analyzed by conventional RT-PCR whereas 30 samples were found to be mixed infection when duplex real-time RT-PCR was applied. This duplex assay provides a useful tool for routine health monitoring and screening of contaminated biological materials of these two viruses.

  1. Simultaneous Determination of Seven Phenolic Acids in Rat Plasma Using UHPLC-ESI-MS/MS after Oral Administration of Echinacea purpurea Extract

    Directory of Open Access Journals (Sweden)

    Yan Du

    2017-09-01

    Full Text Available A rapid and sensitive Ultra High Performance Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (UHPLC-ESI-MS/MS method was developed and validated to simultaneously determine the concentration of seven phenolic acids (syringic acid, ferulic acid, caffeic acid, vanillic acid, p-coumaric acid, 3,4-dihydroxybenzoic acid and 4-hydroxybenzoic acid in rat plasma after oral administration of Echinacea purpurea extract. After mixing with the internal standard (IS, butylparaben, plasma samples were prepared by liquid–liquid extraction with ethyl acetate. The separation was performed using the Agilent Eclipse Plus C18 column (1.8 μm, 2.1 mm × 50 mm with a gradient system consisting of solution A (0.1% acetic acid in water and solution B (methanol at a flow rate of 0.3 mL/min. The detection was accomplished by a multiple reaction monitoring (MRM mode with electrospray ionization (ESI. The method was validated in terms of linearity, precision, accuracy, extraction recovery, matrix effect and stability. This method was successfully applied to study the pharmacokinetic properties of the seven compounds after oral administration of Echinacea purpurea extract in rats.

  2. Simultaneous Determination of Seven Phenolic Acids in Rat Plasma Using UHPLC-ESI-MS/MS after Oral Administration of Echinacea purpurea Extract.

    Science.gov (United States)

    Du, Yan; Wang, Zhibin; Wang, Libo; Gao, Mingjie; Wang, Liqian; Gan, Chunli; Yang, Chunjuan

    2017-09-07

    A rapid and sensitive Ultra High Performance Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (UHPLC-ESI-MS/MS) method was developed and validated to simultaneously determine the concentration of seven phenolic acids (syringic acid, ferulic acid, caffeic acid, vanillic acid, p-coumaric acid, 3,4-dihydroxybenzoic acid and 4-hydroxybenzoic acid) in rat plasma after oral administration of Echinacea purpurea extract. After mixing with the internal standard (IS), butylparaben, plasma samples were prepared by liquid-liquid extraction with ethyl acetate. The separation was performed using the Agilent Eclipse Plus C18 column (1.8 μm, 2.1 mm × 50 mm) with a gradient system consisting of solution A (0.1% acetic acid in water) and solution B (methanol) at a flow rate of 0.3 mL/min. The detection was accomplished by a multiple reaction monitoring (MRM) mode with electrospray ionization (ESI). The method was validated in terms of linearity, precision, accuracy, extraction recovery, matrix effect and stability. This method was successfully applied to study the pharmacokinetic properties of the seven compounds after oral administration of Echinacea purpurea extract in rats.

  3. Data supporting the rat brain sample preparation and validation assays for simultaneous determination of 8 neurotransmitters and their metabolites using liquid chromatography–tandem mass spectrometry

    Science.gov (United States)

    Wojnicz, Aneta; Ortiz, José Avendaño; Casas, Ana I.; Freitas, Andiara E.; López, Manuela G.; Ruiz-Nuño, Ana

    2016-01-01

    The data presented in this article supports the rat brain sample preparation procedure previous to its injection into the liquid chromatography–tandem mass spectrometry (LC–MS/MS) system to monitor levels of adrenaline, noradrenaline, glutamic acid, γ-aminobutyric acid, dopamine, 5-hydroxytryptamine, 5-hydroxyindole acetic acid, and 3-methoxy-4-hydroxyphenylglycol. In addition, we describe the method validation assays (such as calibration curve, lower limit of quantification, precision and accuracy intra- and inter-day, selectivity, extraction recovery and matrix effect, stability, and carry-over effect) according to the United States Food and Drug Administration and European Medicine Agency to measure in one step different neurotransmitters and their metabolites. The data supplied in this article is related to the research study entitled: “Simultaneous determination of 8 neurotransmitters and their metabolite levels in rat brain using liquid chromatography in tandem with mass spectrometry: application to the murine Nrf2 model of depression” (Wojnicz et al. 2016) [1]. PMID:27054183

  4. Simultaneous Determination of Typhaneoside and Isorhamnetin-3-O-Neohesperidoside in Rats After Oral Administration of Pollen Typhae Extract by UPLC-MS/MS.

    Science.gov (United States)

    Cao, Sali; Ni, Boran; Feng, Lijun; Yin, Xingbin; Dou, Haoran; Fu, Jing; Lin, Longfei; Ni, Jian

    2015-07-01

    For the first time, a selective and rapid ultra-performance liquid chromatography method with tandem mass spectrometric (UPLC-MS/MS) detection for simultaneous determination of typhaneoside and isorhamnetin-3-O-neohesperidoside in rat plasma was developed and validated, which was applied to the pharmacokinetic study of Pollen Typhae extract. The separation was carried out on an ACQUITY UPLC(TM) BEH C18 column with gradient elution using mobile phase including acetonitrile and water (containing 0.1% formic acid). The flow rate was 0.4 mL/min. The detection was conducted by means of electrospray ionization mass spectrometry in negative ion mode with multiple reaction monitoring. The assays were linear over the concentration range of 0.5-100 ng/mL, and the lower limit of quantification was 0.5 ng/mL for typhaneoside and isorhamnetin-3-O-neohesperidoside. The method was validated in terms of intra- and interday precision (<9.37%), accuracy (within ±10.91%), linearity, specificity and stability, and has been successfully applied to a pharmacokinetic study of Pollen Typhae extract in rats after oral administration.

  5. Simultaneous determination of naringin, hesperidin, neohesperidin, naringenin and hesperetin of Fractus aurantii extract in rat plasma by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Tong, Ling; Zhou, Dandan; Gao, Jun; Zhu, Yonghong; Sun, He; Bi, Kaishun

    2012-01-25

    A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of naringin, hesperidin, neohesperidin, naringenin and hesperetin in rat plasma, using liquiritin as the internal standard. Plasma samples extracted with a solid-phase extraction procedure were separated on a Zorbax SB-C18 analytical column (2.1 mm × 150 mm, 5 μm) and detected by electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curves were linear over the range of 3.0-600 ng/ml for naringin, 0.5-100 ng/ml for hesperidin, 3.5-700 ng/ml for neohesperidin, 5.0-1000 ng/ml for naringenin and hesperetin, respectively. The lower limits of quantification were 0.5 ng/ml for naringin, hesperidin, naringenin and hesperetin, and 0.35 ng/ml for neohesperidin. Intra- and inter-day precision (RSD%) was less than 15% and accuracy (RE%) ranged from -3.3% to 4.8%. The validated method was successfully applied to investigate the pharmacokinetics of the major flavanones of Fructus aurantii extract after oral administration to rats.

  6. Dipole source analysis for readiness potential and field using simultaneously measured EEG and MEG signals.

    Science.gov (United States)

    Mideksa, K G; Hellriegel, H; Hoogenboom, N; Krause, H; Schnitzler, A; Deuschl, G; Raethjen, J; Heute, U; Muthuraman, M

    2013-01-01

    Various source localization techniques have indicated the generators of each identifiable component of movement-related cortical potentials, since the discovery of the surface negative potential prior to self-paced movement by Kornhuber and Decke. Readiness potentials and fields preceding self-paced finger movements were recorded simultaneously using multichannel electroencephalography (EEG) and magnetoencephalography (MEG) from five healthy subjects. The cortical areas involved in this paradigm are the supplementary motor area (SMA) (bilateral), pre-SMA (bilateral), and contralateral motor area of the moving finger. This hypothesis is tested in this paper using the dipole source analysis independently for only EEG, only MEG, and both combined. To localize the sources, the forward problem is first solved by using the boundary-element method for realistic head models and by using a locally-fitted-sphere approach for spherical head models consisting of a set of connected volumes, typically representing the scalp, skull, and brain. In the source reconstruction it is to be expected that EEG predominantly localizes radially oriented sources while MEG localizes tangential sources at the desired region of the cortex. The effect of MEG on EEG is also observed when analyzing both combined data. When comparing the two head models, the spherical and the realistic head models showed similar results. The significant points for this study are comparing the source analysis between the two modalities (EEG and MEG) so as to assure that EEG is sensitive to mostly radially orientated sources while MEG is only sensitive to only tangential sources, and comparing the spherical and individual head models.

  7. Precise simultaneous quantification of methadone and cocaine in rat serum and brain tissue samples following their successive i.p. administration.

    Science.gov (United States)

    Nakhla, David S; Hussein, Lobna A; Magdy, N; Abdallah, Inas A; Hassan, Hazem E

    2017-03-24

    A sensitive high-performance liquid chromatography (HPLC) assay with dual UV detection has been developed and validated for the simultaneous quantification of methadone and cocaine in rat serum and brain tissue samples. Liquid-liquid extraction using hexanes was applied for samples extraction with Levo-Tetrahydropalmatine (L-THP) as the internal standard. Chromatographic separation of the analytes was achieved on a reversed-phase Waters Symmetry(®) C18 column (150mm×4.6mm, 5μm). A gradient elution was employed with a mobile phase consisting of 5mM potassium phosphate containing 0.1% triethylamine (pH=6.5) (A) and acetonitrile (B) with a flow rate of 1mL/min. UV detection was employed at 215nm and 235nm for the determination of methadone and cocaine, respectively. The calibration curves were linear over the range of 0.05-10μg/mL for both methadone and cocaine. The assay was validated according to FDA guidelines for bioanalytical method validation and results were satisfactory and met FDA criteria. Inter-day accuracy values of serum and brain samples ranged from 96.97 to 105.59% while intra-day accuracy values ranged from 91.49 to 111.92%. Stability assays showed that both methadone and cocaine were stable during sample storage, preparation, and analytical procedures. The method was successfully used to analyze biological samples obtained from a drug- drug interaction pharmacokinetics (PK) study conducted in rats to investigate the effect of methadone on cocaine PK. Our method not only can be used for bioanalysis of samples obtained from rats but also can potentially be applied to human biological serum samples to monitor compliance to methadone maintenance therapy (MMT) and to detect possible cocaine-methadone co-abuse.

  8. Enantioselective separation and simultaneous determination of tolperisone and eperisone in rat plasma by LC-MS/MS.

    Science.gov (United States)

    Nageswara Rao, R; Satyanarayana Raju, S

    2013-10-01

    Tolperisone and eperisone used as muscle relaxants possess one chiral center each and exist as two optical isomers for each drug. Therefore, enantioselective assays to measure each enantiomer in biological matrices are of great importance. In the present study a simple and complete reverse-phase liquid chromatography tandem mass spectrometric method for separation and enantioselective determination of tolperisone and eperisone in rat plasma was developed. The analytes were extracted from rat plasma by a simple protein precipitation method with acetonitrile as the extraction solvent. The enantioselective separation of analytes was achieved on a Cellulose Tris (4-chloro-3-methylphenylcarbamate) chiral column with a mobile phase of acetonitrile: 10 mM ammonium acetate in an isocratic mode of elution and mass spectrometric detection. The calibration curve for each enantiomer was found to be linear over 0.2 to 20 ng/mL for each enantiomer. The proposed method exhibited good intra- and interday precision (% CV) ranged between 0.95-6.05% and 1.11-8.21%, respectively. The intra- and interday accuracy for the proposed assay method ranged between 94.0-100.5% and 92.7-102.1%, respectively. The proposed method was validated as per regulatory guidelines.

  9. Simultaneous Analysis of Nitrogen, Carbon and Sulfur Stable Isotopes and Concentrations in Organics and Soils

    Science.gov (United States)

    Mambelli, S.; Brooks, P. D.; Sutka, R.; Hughes, S.; Finstad, K. M.; Pakes, M. J.; Dawson, T. E.

    2014-12-01

    To date, analysis of diet, food web complexities, biogeochemical cycles, and ecosystem functioning have largely focused on using variation in carbon (C) and nitrogen (N) stable isotope ratios. This is because a great deal is understood about what leads to this variation and because the dual stable isotope analysis of these two elements using continuous flow isotope ratio mass spectrometry (IRMS) is now commonplace. However, the aforementioned studies may all greatly benefit from the additional information one can get from also having sulfur (S) stable isotopes ratio data. Until very recently the analysis of δ34S has traditionally required an additional and often more difficult analytical procedure. Here, we report on the development of a new method that simultaneously analyzes the elemental and isotopic composition of N, C and S in a single sample. The new commercially available instrument includes a modified NCS elemental analyzer in line with an IRMS outfitted with 100 volt AD converters for wide dynamic range. We tested, and modified, this instrument to achieve maximum accuracy and precision for the isotopic measurements of all three elements. We found that the original design needed improvements to achieve our goals by: a) including a component (originally designed for trapping water) as buffer to reduce S memory and obtain reliable δ34S analysis; b) adding an external furnace for complete reduction of nitrogen oxides to N2 gas for accurate δ15N; c) adding a magnesium perchlorate water trap immediately after the reduction tube to minimize any water condensation that could also influence S memory. We analyzed a selection of organic materials and soils with approximately a 1:2 standards versus unknowns ratio per run. Using this NCS set-up, the precision of the N and C isotopic measurements was comparable to the one usually attained in NC mode alone (standard deviation of ± 0.13 δ15N in the range 30 to 400 µg N, and of ± 0.12 δ13C in the range 0.20 to 4 mg

  10. Simultaneous stable isotope analysis of methane and nitrous oxide on ice core samples

    Directory of Open Access Journals (Sweden)

    C. J. Sapart

    2011-12-01

    Full Text Available Methane and nitrous oxide are important greenhouse gases which show a strong increase in atmospheric mixing ratios since pre-industrial time as well as large variations during past climate changes. The understanding of their biogeochemical cycles can be improved using stable isotope analysis. However, high-precision isotope measurements on air trapped in ice cores are challenging because of the high susceptibility to contamination and fractionation.

    Here, we present a dry extraction system for combined CH4 and N2O stable isotope analysis from ice core air, using an ice grating device. The system allows simultaneous analysis of δD(CH4 or δ13C(CH4, together with δ15N(N2O, δ18O(N2O and δ15N(NO+ fragment on a single ice core sample, using two isotope mass spectrometry systems. The optimum quantity of ice for analysis is about 600 g with typical "Holocene" mixing ratios for CH4 and N2O. In this case, the reproducibility (1σ is 2.1‰ for δD(CH4, 0.18‰ for δ13C(CH4, 0.51‰ for δ15N(N2O, 0.69‰ for δ18O(N2O and 1.12‰ for δ15N(NO+ fragment. For smaller amounts of ice the standard deviation increases, particularly for N2O isotopologues. For both gases, small-scale intercalibrations using air and/or ice samples have been carried out in collaboration with other institutes that are currently involved in isotope measurements of ice core air. Significant differences are shown between the calibration scales, but those offsets are consistent and can therefore be corrected for.

  11. Simultaneous determination of five free and total flavonoids in rat plasma by ultra HPLC-MS/MS and its application to a comparative pharmacokinetic study in normal and hyperlipidemic rats.

    Science.gov (United States)

    Wang, Xiaofan; Zhao, Xu; Gu, Liqiang; Lv, Chunxiao; He, Bosai; Liu, Zhenzhen; Hou, Pengyi; Bi, Kaishun; Chen, Xiaohui

    2014-03-15

    A simple and rapid ultra-high performance liquid chromatography-tandem mass spectrometry (uHPLC-MS/MS) method has been developed for the simultaneous determination of five free flavonoids (amentoflavone, isorhamnetin, naringenin, kaempferol and quercetin) and their total (free and conjugated) forms, and to compare the pharmacokinetics of these active ingredients in normal and hyperlipidemic rats. The free and total forms of these flavonoids were extracted by liquid-liquid extraction with ethyl acetate. The conjugated flavonoids were deconjugated by the enzyme β-Glucuronidase and Sulfatase. Chromatographic separation was accomplished on a ZORBAX Eclipse XDB-C8 USP L7 column using gradient elution. Detection was performed on a 4000Q uHPLC-MS/MS system from AB Sciex using negative ion mode in the multiple reaction monitoring (MRM) mode. The lower limits of quantification were 2.0-5.0ng/mL for all the analytes. Intra-day and inter-day precision were less than 15% and accuracy ranged from -9.3% to 11.0%, and the mean extraction recoveries of analytes and internal standard (IS) from rat plasma were all more than 81.7%. The validated method was successfully applied to a comparative pharmacokinetic study of five free and total analytes in rat plasma. The results indicated that the absorption of five total flavonoids in hyperlipidemia group were significantly higher than those in normal group with similar concentration-time curves.

  12. Rapid Multisample Analysis for Simultaneous Determination of Anthropogenic Radionuclides in Marine Environment

    DEFF Research Database (Denmark)

    Qiao, Jixin; Shi, Keliang; Hou, Xiaolin

    2014-01-01

    An automated multisample processing flow injection (FI) system was developed for simultaneous determination of technetium, neptunium, plutonium, and uranium in large volume (200 L) seawater. Ferrous hydroxide coprecipitation was used for the preliminary sample treatment providing the merit...

  13. Intelligent Simultaneous Quantitative Online Analysis of Environmental Trace Heavy Metals with Total-Reflection X-Ray Fluorescence

    Directory of Open Access Journals (Sweden)

    Junjie Ma

    2015-05-01

    Full Text Available Total-reflection X-ray fluorescence (TXRF has achieved remarkable success with the advantages of simultaneous multi-element analysis capability, decreased background noise, no matrix effects, wide dynamic range, ease of operation, and potential of trace analysis. Simultaneous quantitative online analysis of trace heavy metals is urgently required by dynamic environmental monitoring and management, and TXRF has potential in this application domain. However, it calls for an online analysis scheme based on TXRF as well as a robust and rapid quantification method, which have not been well explored yet. Besides, spectral overlapping and background effects may lead to loss of accuracy or even faulty results during practical quantitative TXRF analysis. This paper proposes an intelligent, multi-element quantification method according to the established online TXRF analysis platform. In the intelligent quantification method, collected characteristic curves of all existing elements and a pre-estimated background curve in the whole spectrum scope are used to approximate the measured spectrum. A novel hybrid algorithm, PSO-RBFN-SA, is designed to solve the curve-fitting problem, with offline global optimization and fast online computing. Experimental results verify that simultaneous quantification of trace heavy metals, including Cr, Mn, Fe, Co, Ni, Cu and Zn, is realized on the online TXRF analysis platform, and both high measurement precision and computational efficiency are obtained.

  14. Intelligent simultaneous quantitative online analysis of environmental trace heavy metals with total-reflection X-ray fluorescence.

    Science.gov (United States)

    Ma, Junjie; Wang, Yeyao; Yang, Qi; Liu, Yubing; Shi, Ping

    2015-05-06

    Total-reflection X-ray fluorescence (TXRF) has achieved remarkable success with the advantages of simultaneous multi-element analysis capability, decreased background noise, no matrix effects, wide dynamic range, ease of operation, and potential of trace analysis. Simultaneous quantitative online analysis of trace heavy metals is urgently required by dynamic environmental monitoring and management, and TXRF has potential in this application domain. However, it calls for an online analysis scheme based on TXRF as well as a robust and rapid quantification method, which have not been well explored yet. Besides, spectral overlapping and background effects may lead to loss of accuracy or even faulty results during practical quantitative TXRF analysis. This paper proposes an intelligent, multi-element quantification method according to the established online TXRF analysis platform. In the intelligent quantification method, collected characteristic curves of all existing elements and a pre-estimated background curve in the whole spectrum scope are used to approximate the measured spectrum. A novel hybrid algorithm, PSO-RBFN-SA, is designed to solve the curve-fitting problem, with offline global optimization and fast online computing. Experimental results verify that simultaneous quantification of trace heavy metals, including Cr, Mn, Fe, Co, Ni, Cu and Zn, is realized on the online TXRF analysis platform, and both high measurement precision and computational efficiency are obtained.

  15. CHARACTERIZATION OF THE DA-ERGIC SYSTEM IN THE MEDIOBASAL HYPOTHALAMUS - A NEW APPROACH TO SIMULTANEOUSLY MONITOR THE RELEASE OF DA FROM THE TIDA NEURONS AND THE PRL SECRETION FROM THE ADENOHYPOPHYSIS IN AWAKE RATS

    NARCIS (Netherlands)

    TIMMERMAN, W; DEINUM, ME; POELMAN, RT; WESTERINK, BHC; SCHUILING, GA

    1994-01-01

    The TIDA neurons, which constitute part of the arcuate nucleus-ME complex, play an important inhibitory role in the regulation of the PRL secretion from the adenohypophysis. To simultaneously study the release of DA from the TIDA neurons and the PRL secretion from the adenohypophysis in awake rats,

  16. A new estimate of afrotherian phylogeny based on simultaneous analysis of genomic, morphological, and fossil evidence

    Directory of Open Access Journals (Sweden)

    Seiffert Erik R

    2007-11-01

    Full Text Available Abstract Background The placental mammalian clade Afrotheria is now supported by diverse forms of genomic data, but interordinal relationships within, and morphological support for, the group remains elusive. As a means for addressing these outstanding problems, competing hypotheses of afrotherian interordinal relationships were tested through simultaneous parsimony analysis of a large data set (> 4,590 parsimony informative characters containing genomic data (> 17 kb of nucleotide data, chromosomal associations, and retroposons and 400 morphological characters scored across 16 extant and 35 extinct afrotherians. Results Parsimony analysis of extant taxa alone recovered the interordinal topology (Afrosoricida, ((Macroscelidea, Tubulidentata, (Hyracoidea, (Proboscidea, Sirenia. Analysis following addition of extinct taxa instead supported Afroinsectivora (Afrosoricida + Macroscelidea and Pseudoungulata (Tubulidentata + Paenungulata, as well as Tethytheria (Proboscidea + Sirenia. This latter topology is, however, sensitive to taxon deletion and different placements of the placental root, and numerous alternative interordinal arrangements within Afrotheria could not be statistically rejected. Relationships among extinct stem members of each afrotherian clade were more stable, but one alleged stem macroscelidean (Herodotius never grouped with that clade and instead consistently joined pseudoungulates or paenungulates. When character transformations were optimized onto a less resolved afrotherian tree that reflects uncertainty about the group's interordinal phylogeny, a total of 21 morphological features were identified as possible synapomorphies of crown Afrotheria, 9 of which optimized unambiguously across all character treatments and optimization methods. Conclusion Instability in afrotherian interordinal phylogeny presumably reflects rapid divergences during two pulses of cladogenesis – the first in the Late Cretaceous, at and just after the

  17. Simultaneous analysis of hemoglobin adducts of acrylamide and glycidamide by gas chromatography-mass spectrometry.

    Science.gov (United States)

    Pérez, H L; Cheong, H K; Yang, J S; Osterman-Golkar, S

    1999-10-01

    Acrylamide (AA) is a carcinogen in experimental animals. Glycidamide (GA), formed by metabolic epoxidation of AA, is believed to be responsible for the carcinogenicity of AA. Occupational exposure to AA has been assessed earlier by measurement of its adducts with N-terminal valine in hemoglobin. A background of AA adducts [N-(2-carbamoylethyl)valine (AAVal), about 30 pmol/g globin] was found in individuals without known exposure to the compound. The method previously available for adducts of GA only allowed analysis of samples from highly exposed individuals and showed similar levels of AAVal and adducts of GA [N-(2-hydroxy-2-carbamoylethyl)valine (GAVal)]. We have developed a sensitive method for simultaneous quantification of adducts of GA and AA, which is suitable down to low exposure levels. The method is based on the so-called modified Edman method, where globin is reacted with pentafluorophenyl isothiocyanate under neutral conditions. The valine adducts are then extracted in the form of pentafluorophenylthiohydantoin (PFPTH) derivatives. The analytical procedure included reaction of the PFPTH derivatives with acetic anhydride in order to protect the hydroxyl group of GAVal. The PFPTH derivatives of AAVal and GAVal were analyzed by gas chromatography/tandem mass spectrometry. ((2)H(3))AAVal-PFPTH was used as the internal standard. The method was applied to samples from 11 workers at an AA production plant, 1 nonexposed nonsmoker, and a few participants of a smoking cessation program. AAVal levels were in the range 27-1854 pmol/g globin. Recorded levels of GAVal were 3-12% of those of AAVal, suggesting that previous measurements of GAVal overestimate GAVal at low levels of exposure to AA.

  18. Simultaneous analysis of all SNPs in genome-wide and re-sequencing association studies.

    Directory of Open Access Journals (Sweden)

    Clive J Hoggart

    2008-07-01

    Full Text Available Testing one SNP at a time does not fully realise the potential of genome-wide association studies to identify multiple causal variants, which is a plausible scenario for many complex diseases. We show that simultaneous analysis of the entire set of SNPs from a genome-wide study to identify the subset that best predicts disease outcome is now feasible, thanks to developments in stochastic search methods. We used a Bayesian-inspired penalised maximum likelihood approach in which every SNP can be considered for additive, dominant, and recessive contributions to disease risk. Posterior mode estimates were obtained for regression coefficients that were each assigned a prior with a sharp mode at zero. A non-zero coefficient estimate was interpreted as corresponding to a significant SNP. We investigated two prior distributions and show that the normal-exponential-gamma prior leads to improved SNP selection in comparison with single-SNP tests. We also derived an explicit approximation for type-I error that avoids the need to use permutation procedures. As well as genome-wide analyses, our method is well-suited to fine mapping with very dense SNP sets obtained from re-sequencing and/or imputation. It can accommodate quantitative as well as case-control phenotypes, covariate adjustment, and can be extended to search for interactions. Here, we demonstrate the power and empirical type-I error of our approach using simulated case-control data sets of up to 500 K SNPs, a real genome-wide data set of 300 K SNPs, and a sequence-based dataset, each of which can be analysed in a few hours on a desktop workstation.

  19. Simultaneous determination of Eleutheroside B and Eleutheroside E in rat plasma by high performance liquid chromatography-electrospray ionization mass spectrometry and its application in a pharmacokinetic study.

    Science.gov (United States)

    Ma, Bo; Zhang, Qi; Liu, Yinhui; Li, Jing; Xu, Qiuyu; Li, Xiaotian; Yang, Xiaojing; Yao, Di; Sun, Jingjing; Cui, Guangbo; Ying, Hanjie

    2013-02-15

    Eleutheroside B and Eleutheroside E, two kinds of the major bioactive saponins of Eleutherococcus senticosus, play a pivotal role in biologic activity. In this study, a specific and sensitive high performance liquid chromatography-electrospray ionization-tandem mass spectrometry method (HPLC-MS/MS) was developed and validated for simultaneous determination of Eleutheroside B and Eleutheroside E in rat plasma. The analytes were extracted from rat plasma via a simple protein precipitation procedure with methanol and polygonin was used as internal standard. Chromatographic separation was achieved on a C18 column using a gradient elution program with acetonitrile and water containing 0.1% ammonium hydroxide solution as the mobile phase, with a flow rate of 0.2mL/min. The detection was performed on a triple-quadrupole tandem mass spectrometer by multiple reactions monitoring (MRM) mode in a negative ion mode via electrospray ionization (ESI). The transition monitored were m/z 371 [M-H](-)→209 for Eleutheroside B, m/z 741[M-H](-)→579 for Eleutheroside E and m/z 389[M-H](-)→277 for internal standard. Linear calibration curves were obtained in the concentration range of 1-2000ng/mL for both (Eleutheroside B and Eleutheroside E), with a lower limit of quantification of 1ng/mL. Extraction recovery was over 80% in plasma. The intra- and inter-day precision (RSD) values were below 12% and accuracy (RE) was -2.80 to 5.70% at three QC levels for both. The assay was successfully applied to study pharmacokinetics behavior in rats after oral and intravenous administration of the single substances (Eleutheroside B and Eleutheroside E). And further research was performed by comparing the difference in pharmacokinetic behavior between the single substances and an aqueous extract of E. senticosus after oral administration. Significant difference in pharmacokinetic characteristics between the single substances and an aqueous extract was found in rat, which would be beneficial for

  20. Serum lipidomics analysis of ovariectomized rats under Curcuma comosa treatment.

    Science.gov (United States)

    Vinayavekhin, Nawaporn; Sueajai, Jetjamnong; Chaihad, Nichaboon; Panrak, Ratchanee; Chokchaisiri, Ratchanaporn; Sangvanich, Polkit; Suksamrarn, Apichart; Piyachaturawat, Pawinee

    2016-11-04

    Curcuma comosa Roxb. (C. comosa) or Wan Chak Motluk, Zingiberaceae family, has been used in Thai traditional medicine for the treatment of gynecological problems and inflammation. This study aimed to investigate the therapeutic potential of C. comosa by determining the changes in the lipid profiles in the ovariectomized rats, as a model of estrogen-deficiency-induced hyperlipidemia, after treatment with different components of C. comosa using an untargeted lipidomics approach. Lipids were extracted from the serum of adult female rats subjected to a sham operation (SHAM; control), ovariectomy (OVX), or OVX with 12-week daily doses of estrogen (17β-estradiol; E2), (3R)-1,7-diphenyl-(4E,6E)-4,6-heptadien-3-ol (DPHD; a phytoestrogen from C. comosa), powdered C. comosa rhizomes or its crude ethanol extract. They were then analyzed by liquid chromatography-mass spectrometry, characterized, and subjected to the orthogonal projections to latent structures discriminant analysis statistical model to identify tentative biomarkers. Levels of five classes of lipids (ceramide, ceramide-1-phosphate, sphingomyelin, 1-O-alkenyl-lysophosphatidylethanolamine and lysophosphatidylethanolamine) were elevated in the OVX rats compared to those in the SHAM rats, while the monoacylglycerols and triacylglycerols were decreased. The E2 treatment only reversed the levels of ceramides, whereas treatments with DPHD, C. comosa extract or powder returned the levels of all upregulated lipids back to those in the SHAM control rats. The findings suggest the potential beneficial effects of C. comosa on preventing the increased ceramide levels in OVX rats, a possible cause of metabolic disturbance under estrogen deficiency. Overall, the results demonstrated the power of untargeted lipidomics in discovering disease-relevant biomarkers, as well as evaluating the effectiveness of treatment by C. comosa components (DPHD, extract or powder) as utilized in Thai traditional medicine, and also providing

  1. Simultaneous determination of five naphthoylindole-based synthetic cannabinoids and metabolites and their deposition in human and rat hair.

    Science.gov (United States)

    Kim, Jihyun; Park, Yuran; Park, Meejung; Kim, Eunmi; Yang, Wonkyung; Baeck, Seungkyung; Lee, Sooyeun; Han, Sangbeom

    2015-01-01

    The continuing appearance of new synthetic cannabinoids has been a major issue in the field of forensic and clinical toxicology. In response to that, analytical methods for synthetic cannabinoids have been increasingly established in a variety of biological matrices. Since most of synthetic cannabinoids with structure similarity share some enzymatic metabolites, making the interpretation of analytical results and the discovery of the parent drug actually ingested very complicated, the investigation on metabolites of the first generation of synthetic cannabinoids with their relatively short side chains in chemical structure could be more important. Therefore, in the present study, we developed the analytical method for AM-2201, JWH-122 and MAM-2201 with JWH-018 as a precursor and their monohydroxylated metabolites in hair matrix. Also, using a rat model, AM-2201 and its monohydroxylated metabolites were identified and then the ratios of metabolite-to-parent drug were estimated to be used as criteria on external contamination. All analytes were extracted with methanol from washed and cut hair samples and the extracts were injected into LC-MS/MS with electrospray ion source in the positive ionization mode. Matrix effect and recovery were evaluated in hair matrices and no significant variations were observed. The validation results for precision and accuracy were satisfactory in both human and rat hair. The LOD and LOQ were 0.5 pg/10mg and 1.0 pg/10mg in human hair and 0.5 pg/20mg and 1.0 pg/20mg in pigmented and non-pigmented rat hair, respectively. Additionally, as a result of the animal study, there were not significant differences in the effect of pigmentation on the distribution of AM-2201 and its monohydroxylated metabolites in hair. Wide variations were observed for the concentrations of the naphthoylindole-based synthetic cannabinoids and metabolites in authentic hair samples from nine cases; those were 0.4-59.2 pg/mg for JWH-018, 0.1-0.8 pg/mg for JWH-073, 1

  2. One-step Real-time Food Quality Analysis by Simultaneous DSC-FTIR Microspectroscopy.

    Science.gov (United States)

    Lin, Shan-Yang; Lin, Chih-Cheng

    2016-01-01

    This review discusses an analytical technique that combines differential scanning calorimetry and Fourier-transform infrared (DSC-FTIR) microspectroscopy, which simulates the accelerated stability test and detects decomposition products simultaneously in real time. We show that the DSC-FTIR technique is a fast, simple and powerful analytical tool with applications in food sciences. This technique has been applied successfully to the simultaneous investigation of: encapsulated squid oil stability; the dehydration and intramolecular condensation of sweetener (aspartame); the dehydration, rehydration and solidification of trehalose; and online monitoring of the Maillard reaction for glucose (Glc)/asparagine (Asn) in the solid state. This technique delivers rapid and appropriate interpretations with food science applications.

  3. Microsurgical technique of simultaneous pancreas/kidney transplantation in the rat: clinical experience and review of the literature.

    Science.gov (United States)

    Matevossian, E; Doll, D; Sinicina, I; Kern, H; Bald, C; Nährig, J; Stangl, M; Thorban, S; Hüser, N

    2009-01-01

    For experimental basic research, standardized transplantation models reflecting technical and immunologic aspects are necessary. This article describes an experimental model of combined pancreas/kidney transplantation (PKTx) in detail. Donor rats underwent en bloc pancreatectomy and nephrectomy. Revascularization was performed using the aorta with the superior mesenteric artery and the inferior vena cava with the portal vein. Exocrine drainage of the pancreas took place over a segment of the duodenum which was transplanted side-to-side to the jejunum. The kidney vessels were transplanted end-to-side. The ureter was anastomosed by patch technique. Postoperatively, serum parameters were monitored daily. Biopsies for histopathology were taken on days 5, 8 and 12. All 12 recipients survived the combined PKTx without serious surgical complications. One thrombosis of the portal vein led to organ failure. Blood glucose levels were normal by the 3rd postoperative day. The transplanted duodenal segment showed slight villous atrophy, and the kidneys were well perfused without vascular complications. The anastomosis between ureter and bladder was leakproof. Excellent graft function and survival rates can be achieved due to simplified operation technique and short operation time. It may thus have high clinical relevance to immunologic issues within the scope of basic research. Copyright 2009 S. Karger AG, Basel.

  4. Simultaneous determination of amlodipine, valsartan and hydrochlorothiazide by LC–ESI-MS/MS and its application to pharmacokinetics in rats

    Directory of Open Access Journals (Sweden)

    Shankar Ganesh Gadepalli

    2014-12-01

    Full Text Available Polypill is a fixed-dose combination that contains three or more active ingredients used as a single daily pill to achieve a large effect in preventing cardiovascular disease with minimal adverse effects. A novel and accurate liquid chromatography tandem mass spectrometry method using electrospray ionization mode has been developed and validated for the simultaneous determination of amlodipine (AMD, valsartan (VAL using losartan (LOS as an internal standard (IS, and hydrochlorothiazide (HCT using furosemide (FSD as an IS. The separation was carried on Aquasil C18 (50 mm×2.1 mm, 5 µm reversed phase column using acetonitrile and water containing 0.1% formic acid (50:50, v/v as the mobile phase. The method was validated in terms of linearity, accuracy and precision over the concentration range of 1–1000 ng/mL. The intra and inter-day precision and accuracy, stability and extraction recoveries of all the analytes were in the acceptable range. This method can be successfully applied to the pharmacokinetic study of AMD, VAL and HCT when given as a polypill.

  5. Isobolographic analysis of the interaction between fenoldopam and levodopa on arterial blood pressure of the rat.

    Science.gov (United States)

    Quevedo, M; Prieto, J C; Miranda, H F; Pinardi, G

    2000-09-01

    Fenoldopam (FD) and levodopa (LD) injected intravenously in rats in a noncumulative schedule induced dose-dependent reductions in mean arterial blood pressure. The doses that induced a 50% reduction in the initial control mean arterial pressure (referred as ED50) were calculated by linear regression analysis of the corresponding parallel dose-response curves and were 0.88 and 0.068 mg/kg, respectively. The interaction between the effects of FD and LD on pressure reduction was evaluated by simultaneous administration of different fixed ratios of FD and LD (16:1) and obtaining a dose-response curve. An isobolographic analysis was then performed, which showed that the experimental point for the effect of the simultaneous administration of FD and LD was significantly different from the theoretically calculated additive point, denoting supradditivity. It was concluded that the effect of the combination of FD and LD on mean blood pressure reduction was synergic and was probably due to an activation of D1 vascular receptors by both drugs, in conjunction with an activation of beta2 adrenoceptors by LD and a blocking action of FD on postsynaptic alpha1-adrenoceptors.

  6. PyRAT (python radiography analysis tool): overview

    Energy Technology Data Exchange (ETDEWEB)

    Armstrong, Jerawan C [Los Alamos National Laboratory; Temple, Brian A [Los Alamos National Laboratory; Buescher, Kevin L [Los Alamos National Laboratory

    2011-01-14

    PyRAT was developed as a quantitative tool for robustly characterizing objects from radiographs to solve problems such as the hybrid nonlinear inverse problem. The optimization software library that was used is the nonsmooth optimization by MADS algorithm (NOMAD). Some of PyRAT's features are: (1) hybrid nonlinear inverse problem with calculated x-ray spectrum and detector response; (2) optimization based inversion approach with goal of identifying unknown object configurations - MVO problem; (3) using functionalities of Python libraries for radiographic image processing and analysis; (4) using the Tikhonov regularization method of linear inverse problem to recover partial information of object configurations; (5) using a priori knowledge of problem solutions to define feasible region and discrete neighbor for the MVO problem - initial data analysis + material library {yields} a priori knowledge; and (6) using the NOMAD (C++ version) software in the object.

  7. A new look at the simultaneous analysis and design of structures

    Science.gov (United States)

    Striz, Alfred G.

    1994-01-01

    The minimum weight optimization of structural systems, subject to strength and displacement constraints as well as size side constraints, was investigated by the Simultaneous ANalysis and Design (SAND) approach. As an optimizer, the code NPSOL was used which is based on a sequential quadratic programming (SQP) algorithm. The structures were modeled by the finite element method. The finite element related input to NPSOL was automatically generated from the input decks of such standard FEM/optimization codes as NASTRAN or ASTROS, with the stiffness matrices, at present, extracted from the FEM code ANALYZE. In order to avoid ill-conditioned matrices that can be encountered when the global stiffness equations are used as additional nonlinear equality constraints in the SAND approach (with the displacements as additional variables), the matrix displacement method was applied. In this approach, the element stiffness equations are used as constraints instead of the global stiffness equations, in conjunction with the nodal force equilibrium equations. This approach adds the element forces as variables to the system. Since, for complex structures and the associated large and very sparce matrices, the execution times of the optimization code became excessive due to the large number of required constraint gradient evaluations, the Kreisselmeier-Steinhauser function approach was used to decrease the computational effort by reducing the nonlinear equality constraint system to essentially a single combined constraint equation. As the linear equality and inequality constraints require much less computational effort to evaluate, they were kept in their previous form to limit the complexity of the KS function evaluation. To date, the standard three-bar, ten-bar, and 72-bar trusses have been tested. For the standard SAND approach, correct results were obtained for all three trusses although convergence became slower for the 72-bar truss. When the matrix displacement method was used

  8. Simultaneous analysis of multiple Mycobacterium tuberculosis knockdown mutants in vitro and in vivo.

    Directory of Open Access Journals (Sweden)

    Antje Blumenthal

    Full Text Available Mycobacterium tuberculosis (Mtb represents one of the most persistent bacterial threats to human health and new drugs are needed to limit its impact. Conditional knockdown mutants can help validate new drug targets, but the analysis of individual mutants is laborious and time consuming. Here, we describe quantitative DNA tags (qTags and their use to simultaneously analyze conditional Mtb knockdown mutants that allowed silencing the glyoxylate and methylcitrate cycles (via depletion of isocitrate lyase, ICL, the serine protease Rv3671c, and the core subunits of the mycobacterial proteasome, PrcB and PrcA. The impact of gene silencing in multi-strain cultures was determined by measuring the relative abundance of mutant-specific qTags with real-time PCR. This achieved accurate quantification over a broad range of qTag abundances and depletion of ICL, Rv3671c, or PrcBA resulted in the expected impairment of growth of Mtb with butyrate as the primary carbon source, survival during oxidative stress, acid stress and starvation. The impact of depleting ICL, Rv3671c, or PrcBA in multi-strain mouse infections was analyzed with two approaches. We first measured the relative abundance of mutant-specific qTags in total chromosomal DNA isolated from bacteria that were recovered from infected lungs on agar plates. We then developed a two-step amplification procedure, which allowed us to measure the abundances of individual mutants directly in infected lung tissue. Both strategies confirmed that inactivation of Rv3671c and PrcBA severely reduced persistence of Mtb in mice. The multi-strain infections furthermore suggested that silencing ICL not only prevented growth of Mtb during acute infections but also prevented survival of Mtb during chronic infections. Analyses of the ICL knockdown mutant in single-strain infections confirmed this and demonstrated that silencing of ICL during chronic infections impaired persistence of Mtb to the extent that the pathogen

  9. Enunciative analysis of modals in libras-portuguese simultaneous interpretation's corpus

    Directory of Open Access Journals (Sweden)

    Anderson Almeida-Silva

    2015-12-01

    Full Text Available This paper focuses on the work of the sign Language interpreter in the condition of a second speaker within the speech production. These professionals, when interpreting, they look for re-create [actualize] the emanated concepts from the source language into the target language, creating, then, a second enunciative instance. The theoretical basis is situated in the perspective of the theory of enunciation (BENVENISTE, 1989; FLORES, 2008, 2009a, 2009b, 2010 and in the study of the modals in spoken languages and LIBRAS. We aim to find what are the marks left in the discourse by the second speaker of the text, in this case, the interpreter, regarding specifically to the use of modals, and additionally, to analyze their choices for using one or other type of modal (in the target text, which might have been made explicit or implicit in the first enunciative context (source text. In order to reach these goals, sampling was composed by a transcription of a video-recorded speech of a deaf person with the length of 40’ (forty minutes that was being simultaneously interpreted into spoken Portuguese. Six excerpts from the total speech, in which modal’s occurrences were attested within the source or target language, were submitted to the analysis. The main findings are: 1. the tendency of the deontic modality being almost always interpreted in the same way of the source language; 2. the interpreting choices are always guided by enunciative clues that are left in the speech of the first speaker, to which the interpreters must to demonstrate expertise in recognizing these language-specific aspects; 3. in the absence of explicit modals in the speech, the interpreters appears to be sensible to the modal’s notions spread by the whole text’s surface; 4. the temporal omission of epistemic modals with a high degree of certainty is understood as an interpretative strategy and not as an infidelity with the original information; 5. the term “tempo de

  10. Simultaneous determination of harpagoside and cinnamic acid in rat plasma by high-performance liquid chromatography: application to a pharmacokinetic study.

    Science.gov (United States)

    Li, Peifan; Zhang, Yunhui; Xiao, Li; Jin, Xinghua; Yang, Kun

    2007-12-01

    Radix Scrophulariae (xuanshen) is one of the famous Chinese herbal medicines widely used to treat rheumatism, tussis, pharyngalgia, arthritis, constipation, and conjunctival congestion. Harpagoside and cinnamic acid are the main bioactive components of xuanshen. The purpose of this study was to develop an HPLC-UV method for simultaneous determination of harpagoside and cinnamic acid in rat plasma and investigate pharmacokinetic parameters of harpagoside and cinnamic acid after oral administration of xuanshen extract (760 mg kg(-1)). After addition of syringin as internal standard, the analytes were isolated from plasma by liquid-liquid extraction. Separation was achieved on a Kromasil C18 column, and detection was by UV absorption at 272 nm. The described assay was validated in terms of linearity, accuracy, precision, recovery, and limit of quantification according to the FDA validation guidelines. Calibration curves for both analytes were linear with the coefficient of variation (r) for both was greater than 0.999. Accuracy for harpagoside and cinnamic acid ranged from 100.7-103.5% and 96.9-102.9%, respectively, and precision for both analytes were less than 8.5%. The main pharmacokinetic parameters found for harpagoside and cinnamic acid after oral infusion of xuanshen extract were as follows: Cmax 1488.7 +/- 205.9 and 556.8 +/- 94.2 ng mL(-1), Tmax 2.09 +/- 0.31 and (1.48 +/- 0.14 h, AUC(0-24) 10,336.4 +/- 1426.8 and 3653.1 +/- 456.4 ng h mL(-1), AUC(0-infinity) 11,276.8 +/- 1321.4 and 3704.5 +/- 398.8 ng h mL(-1), and t(1/2) 4.9 +/- 1.3 and 2.5 +/- 0.9 h, respectively. These results indicated that the proposed method is simple, selective, and feasible for pharmacokinetic study of radix Scrophulariae extract in rats.

  11. Simultaneous quantification of three active alkaloids from a traditional Chinese medicine Ramulus Mori (Sangzhi) in rat plasma using liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Yang, Shuang; Wang, Baolian; Xia, Xuejun; Li, Xue; Wang, Renyun; Sheng, Li; Li, Dan; Liu, Yuling; Li, Yan

    2015-05-10

    Fagomine, 1-deoxynojirimycin (DNJ) and 1,4-dideoxy-1,4-imino-d-arabinitol (DAB) are the major bioactive constituents in the active fraction of alkaloids from the traditional Chinese medicine mulberry twig (Ramulus Mori, Chinese name Sang Zhi), which has a strong activity on α-glucosidase in vitro and in vivo. A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of DNJ, fagomine and DAB in rat plasma. Plasma samples were prepared using a simple protein precipitation by the addition of 1% volume of Tris and two volumes of methanol-acetonitrile. The analytes and internal standard (IS, miglitol) were chromatographed in an XBridge™ amide column with a gradient mobile phase of acetonitrile-water (0.1% ammonium hydroxide) at a flow rate of 0.7mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) source in positive ion mode by multiple reaction monitoring (MRM) mode. Linear detection responses were obtained for DNJ ranging from 5.00 to 5000.00ng/mL, 10.00 to 2500.00ng/mL for fagomine and DAB. The lower limits of quantification (LLOQs) were 5.00, 10.00, 10.00ng/mL for DNJ, fagomine and DAB, respectively. Intra-day and inter-day precisions (R.S.D.%) were within 10% for three analytes with accuracies (R.E.%) less than 12%. The mean recoveries of analytes were greater than 85%. All analytes were proved to be stable during the sample storage, preparation and analytic procedures. The method was successfully applied to the pharmacokinetic study of the three alkaloids in rats after oral administration of the active fraction of alkaloids from mulberry twig.

  12. Simultaneous determination of six flavonoids from Paulownia tomentosa flower extract in rat plasma by LC-MS/MS and its application to a pharmacokinetic study.

    Science.gov (United States)

    Dai, Bin; Hu, Zhiqiang; Li, Haiyan; Yan, Chong; Zhang, Liwei

    2015-01-26

    A simple, rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of six components including apigenin, quercetin, apigenin-7-O-β-D-glucoside, quercetin-3-O-β-D-glucoside, 3'-methoxyluteolin-7-O-β-D-glucoside, and tricin-7-O-β-D-glucopyranoside in rat plasma using formononetin as the internal standard (IS). The plasma samples were pretreated by a one-step liquid-liquid extraction with dichloromethane. The chromatographic separation was carried out on a ZORBAX SB-Aq column with a gradient mobile phase consisting of acetonitrile and 2mM aqueous ammonium acetate. All analytes and IS were quantitated through electrospray ionization in negative ion multiple reaction monitoring mode. The mass transitions were as follows: m/z 269.1→117.2 for apigenin, m/z 301.2→151.2 for quercetin, m/z 431.3→311.2 for apigenin-7-O-β-D-glucoside, m/z 463.2→300.2 for quercetin-3-O-β-D-glucoside, m/z 461.3→283.1 for 3'-methoxyluteolin-7-O-β-D-glucoside, m/z 491.3→313.1 for tricin-7-O-β-D-glucopyranoside, and m/z 267.2→252.2 for IS, respectively. All calibration curves exhibited good linearity with correlation coefficient (r)>0.995. The intra-day and inter-day precisions (RSD) at three QC levels were both less than 14.0% and the accuracies ranged from 89.8% to 113.8%. The extraction recoveries of six compounds ranged from 82.3% to 92.5%. The validated method was successfully applied to pharmacokinetic study of the six components in male rat plasma after oral administration of Paulownia tomentosa flower extract.

  13. Simultaneous parasympathetic and sympathetic activation reveals altered autonomic control of heart rate, vascular tension and epinephrine release in anaesthetized hypertensive rats

    Directory of Open Access Journals (Sweden)

    Torill eBerg

    2011-11-01

    Full Text Available Sympathetic hyperactivity and parasympathetic insufficiency characterize blood pressure control in genetic hypertension, but is difficult to demonstrate experimentally in anesthetized rats. Here we present a pharmacological approach to activate sympathetic and parasympathetic nerves simultaneously, and identify their contribution. Anaesthetized normotensive (WKY and spontaneously hypertensive rats (SHR were injected i.v. with 4-aminopyridine (4-AP, a voltage-sensitive K+ channel inhibitor. Blood pressure was recorded through a femoral artery catheter, cardiac output and heart rate (HR through an ascending aorta flow probe. Total peripheral vascular resistance (TPVR was calculated. 4-AP induced an immediate, atropine- and hexamethonium-sensitive bradycardia in WKY, and in strains, a subsequent, sustained tachycardia, and norepinephrine but not epinephrine release. The tachycardia was eliminated by reserpine, nadolol or right vagal nerve stimulation, but not adrenalectomy, scopolamine or hexamethonium. 4-AP-induced, atropine-sensitive bradycardia was observed in reserpinized or nadolol-treated SHR, where atropine also increased the late HR-response. 4-AP increased TPVR, transiently in WKY but sustained in SHR. Yohimbine but not phentolamine prevented TPVR down-regulation in WKY. Reserpine, phentolamine and prazosin eliminated the late vasoconstriction in SHR. Plasma epinephrine overflow increased in nadolol-treated SHR. Conclusions: 4-AP activated parasympathetic ganglion transmission and peripheral, sympathetic nerve norepinephrine release. The sympathetic component dominated the HR-response to 4-AP in SHR. α2-adrenceptor-dependent vasodilatation opposed norepinephrine-induced α1-adrenergic vasoconstriction in WKY, but not in SHR. A βAR-activated, probably vagal afferent mechanism, hampered adrenal epinephrine secretion in SHR. Thus, 4-AP exposed mechanisms, which contribute to hypertension, and may allow identification of the factors

  14. Simultaneous analysis of small organic acids and humic acids using high performance size exclusion chromatography

    NARCIS (Netherlands)

    Qin, X.P.; Liu, F.; Wang, G.C.; Weng, L.P.

    2012-01-01

    An accurate and fast method for simultaneous determination of small organic acids and much larger humic acids was developed using high performance size exclusion chromatography. Two small organic acids, i.e. salicylic acid and 2,3-dihydroxybenzoic acid, and one purified humic acid material were used

  15. Simultaneous recording of ATP-sensitive K+ current and intracellular Ca2+ in anoxic rat ventricular myocytes. Effects of glibenclamide.

    Science.gov (United States)

    Russ, U; Englert, H; Schölkens, B A; Gögelein, H

    1996-05-01

    We investigated the temporal relationship between the adenosine triphosphate-sensitive K current (KATP current), hypoxic shortening and Ca accumulation in cardiomyocytes exposed to anoxia or metabolic inhibition. Whole-cell, patch-clamp experiments were performed with nonstimulated isolated rat heart ventricular muscle cells loaded with the Ca-sensitive fluorescent dye 1-[2-(5-carboxyoxazol-2-yl)-6-aminobenzofuran-5-oxy]-2-(2'- amino-5'-methylphenoxy) ethane-N,N,N',N'-tetraacetic acid (fura-2) via the patch pipette. After approximately 8 min anoxia, the KATP current started to rise and reached a maximum of 21.3 +/- 3.7 nA (n = 5, recorded at 0 mV clamp potential) within 1-3 min. At that time hypoxic contracture also occurred. Resting cytoplasmic free calcium (Cai) did not change significantly before hypoxic shortening. After hypoxic contracture, the KATP current decreased and Cai started to rise, reaching about 1 micromol/l. The presence of glibenclamide (10 micromol/l) in the bath reduced the anoxia-induced KATP current by more than 50%, but did not significantly influence the time dependence of current, hypoxic shortening and Cai, or the magnitude of Cai. Metabolic inhibition with 1.5 mmol/l CN resulted in KATP current increase and hypoxic shortening, occurring somewhat earlier than under anoxia, but all other parameters were comparable. In non-patch-clamped cells loaded with fura-2 AM ester and field-stimulated with 1 Hz, 1 micronol/l glibenclamide had no significant effect on the magnitude of the Cai increase caused by exposure of the cells to 1.5 mmol/l CN-. After CN- wash-out in non-patch-clamped cells, Cai declined, oscillated and finally returned to control values. It can be concluded that glibenclamide inhibits anoxia-induced KATP currents only partially and has no significant effect on anoxia-induced rise in resting Cai.

  16. LC/MS/MS analysis of the endogenous dimethyltryptamine hallucinogens, their precursors, and major metabolites in rat pineal gland microdialysate.

    Science.gov (United States)

    Barker, Steven A; Borjigin, Jimo; Lomnicka, Izabela; Strassman, Rick

    2013-12-01

    We report a qualitative liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the simultaneous analysis of the three known N,N-dimethyltryptamine endogenous hallucinogens, their precursors and metabolites, as well as melatonin and its metabolic precursors. The method was characterized using artificial cerebrospinal fluid (aCSF) as the matrix and was subsequently applied to the analysis of rat brain pineal gland-aCSF microdialysate. The method describes the simultaneous analysis of 23 chemically diverse compounds plus a deuterated internal standard by direct injection, requiring no dilution or extraction of the samples. The results demonstrate that this is a simple, sensitive, specific and direct approach to the qualitative analysis of these compounds in this matrix. The protocol also employs stringent MS confirmatory criteria for the detection and confirmation of the compounds examined, including exact mass measurements. The excellent limits of detection and broad scope make it a valuable research tool for examining the endogenous hallucinogen pathways in the central nervous system. We report here, for the first time, the presence of N,N-dimethyltryptamine in pineal gland microdialysate obtained from the rat. Copyright © 2013 John Wiley & Sons, Ltd.

  17. Simultaneous Real-Time Analysis of Bulk and Bottom Cure of Ultraviolet-Curable Inks Using Fourier Transform Infrared Spectroscopy.

    Science.gov (United States)

    Boonen, Hennie A L; Koskamp, Janou A; Theiss, Wolfgang; Iedema, Piet D; Willemse, Robin X E

    2017-01-01

    The curing characteristics of an ultraviolet (UV) ink layer are of utmost importance for the development of UV inks. Measuring either bulk or bottom cure in itself is not new and has been the subject of many articles. In this article, two methods are described based on Fourier transform infrared (FT-IR) spectrometry to measure in real time and simultaneously the bulk and bottom cure of a thin UV ink layer. The procedure consists of applying a thin (10-12 µm) layer of UV-curing ink on an attenuated total reflection (ATR) crystal. The bottom cure is measured with ATR. The bulk cure is measured simultaneously with a reflection analysis (method 1) or a transmission analysis (method 2). With both methods, the bulk and bottom cure can be determined. To overcome problems with the interference in the ATR reflection setup, it is recommended to use the ATR transmission setup.

  18. Simultaneous analysis of acidulants and preservatives in food samples by using capillary zone electrophoresis with indirect UV detection.

    Science.gov (United States)

    Yoshikawa, Kenji; Saito, Shintaro; Sakuragawa, Akio

    2011-08-01

    Capillary zone electrophoresis with indirect UV detection was developed for the simultaneous analysis of acidulants and preservatives in food samples. When a solution of tris (hydroxymethyl) aminomethane, trimellitic acid and poly (vinyl alcohol) was used as the background electrolyte, the nine acidulants and four preservatives listed in the Japanese Food Sanitation Law were detected within 8min. The calibration curves plotted from the peak height of each analyte were linear with a correlation coefficient of 0.99. The relative standard deviations (n=10) of the peak height ranged from 1.2% to 4.7%. The detection limits for these species ranged from 0.6 to 5.3mg/L at a signal-to-noise ratio of three. The method developed method was applied to the simultaneous analysis of acidulants and preservatives in a wide variety of food samples.

  19. Validated UPLC-MS/MS method for simultaneous determination of simvastatin, simvastatin hydroxy acid and berberine in rat plasma: Application to the drug-drug pharmacokinetic interaction study of simvastatin combined with berberine after oral administration in rats.

    Science.gov (United States)

    Liu, Mei; Su, Xianying; Li, Guofei; Zhao, Guilian; Zhao, Limei

    2015-12-01

    A rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method was developed and validated for simultaneous quantification of simvastatin (SV), its metabolite simvastatin hydroxy acid (SVA) and berberine (BBR) in rat plasma. Separation was performed on Poroshell 120 EC-C18 column (4.6×50mm, 2.7μm) using gradient elution by mobile phase containing acetonitrile and 10mM ammonium acetate (pH 4.5). Polarity switch (positive-negative-positive ionization mode) was performed in a total run time of 4.0min. The lower limits of quantification (LLOQ) for SV, SVA and BBR were 0.10, 0.20 and 0.10ng/mL, respectively. The response function was established for concentration range of 0.10-100ng/mL for SV and BBR and 0.20-3000ng/mL for SVA, with a coefficient of correlation of >0.99 for all the compounds. The proposed method was applied to the drug-drug pharmacokinetic interaction study of SV combined with BBR after oral administration in rats.

  20. Simultaneous Organic and Inorganic Analysis of Colored Oriental Lacquerware by Pyrolysis-Gas Chromatography/Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Yoshimi Kamiya

    2015-01-01

    Full Text Available Organic analysis and inorganic analysis are generally based on different physical principles, and for this reason it is difficult to analyze resins and pigments simultaneously. For these reasons, we have performed Py-GC/MS measurements of red-, yellow-, and green-colored lacquer films applied to lacquerware items to assess the feasibility of simultaneously detecting resin ingredients together with certain pigments. We have also compared our findings to the results of SEM-EDS, X-ray fluorescence spectrometry (XRF, and X-ray diffractometry (XRD measurements. XRD analysis yielded molecular-level information (information on binding states regarding mercury (Hg and iron (Fe; however, the information obtained for arsenic (As and sulfur (S was insufficient. In contrast, Py-GC/MS analyses simultaneously yielded molecular-level information on arsenic (As and sulfur (S together with detection of the primary ingredients of the lacquer. For this reason, it shows that several pieces of information is provided easily and quickly when the colored lacquer cultural heritage is measured using the Py-GC/MS method.

  1. A UFLC-MS/MS method with a switching ionization mode for simultaneous quantitation of polygalaxanthone III, four ginsenosides and tumulosic acid in rat plasma: application to a comparative pharmacokinetic study in normal and Alzheimer's disease rats.

    Science.gov (United States)

    Lv, Chunxiao; Li, Qing; Zhang, Yaowen; Sui, Zhenyu; He, Bosai; Xu, Huarong; Yin, Yidi; Chen, Xiaohui; Bi, Kaishun

    2013-08-01

    A fast, sensitive and reliable ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method has been developed and validated for simultaneous quantitation of polygalaxanthone III (POL), ginsenoside Rb1 (GRb1), ginsenoside Rd (GRd), ginsenoside Re (GRe), ginsenoside Rg1 (GRg1) and tumulosic acid (TUM) in rat plasma after oral administration of Kai-Xin-San, which plays an important role for the treatment of Alzheimer's disease (AD). The plasma samples were extracted by liquid-liquid extraction using ethyl acetate-isopropanol (1:1, v/v) with salidrdoside as internal standard (IS). Good chromatographic separation was achieved using gradient elution with the mobile phase consisting of methanol and 0.01% acetic acid in water. The tandem mass spectrometric detection was performed in multiple reaction monitoring mode on 4000Q UFLC-MS/MS system with turbo ion spray source in a negative and positive switching ionization mode. The lower limits of quantification were 0.2-1.5 ng/ml for all the analytes. Both intra-day and inter-day precision and accuracy of analytes were well within acceptance criteria (±15%). The mean absolute extraction recoveries of analytes and IS from rat plasma were all more than 60.0%. The validated method has been successfully applied to comparing pharmacokinetic profiles of analytes in normal and AD rat plasma. The results indicated that no significant differences in pharmacokinetic parameters of GRe, GRg1 and TUM were observed between the two groups, while the absorption of POL and GRd in AD group were significantly higher than those in normal group; moreover, the GRb1 absorbed more rapidly in model group. The different characters of pharmacokinetics might be caused by pharmacological effects of the analytes.

  2. Simultaneous determination of two iridoid glycosides, two anthraquinones and four flavonoid glycosides of Zhi-Zi-Da-Huang decoction in rat plasma by UFLC-MS/MS: application to a comparative pharmacokinetic study in normal and cholestatic liver injury rats.

    Science.gov (United States)

    Zhu, Heyun; Bi, Kaishun; Han, Fei; Guan, Jiao; Tang, Zheng; Chen, Kelin; Zhao, Longshan; Li, Qing; Yin, Ran; Hou, Xiaohong

    2014-06-01

    A selective, sensitive and reliable ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method has been developed for the simultaneous determination of two iridoid glycosides (geniposide and genipin gentiobioside), two anthraquinones (rhein and emodin) and four flavonoid glycosides (isonaringin, naringin, hesperidin and neohesperidin), the major active ingredients of Zhi-Zi-Da-Huang decoction (ZZDHD), in rat plasma using paeoniflorin as internal standard (IS). After liquid-liquid extraction with ethyl acetate-isopropanol (1:1, v/v), separation was achieved on a Shim-pack XR-ODS C18 column (75 mm×3.0 mm, 2.2 μm) using gradient elution with a mobile phase consisting of water (containing 0.1% formic acid) and acetonitrile at a flow rate of 0.4 mL/min. Detection was performed on 4000 QTRAP mass spectrometry equipped with turbo ion spray source in the negative ionization and multiple reaction monitoring (MRM) mode. The intra- and inter-day precisions (as relative standard deviation) were less than 11.4%, and accuracy (as relative error) was within ± 10.0%. The lower limits of quantification (LLOQ) were 4.0, 0.5, 2.0, 0.1, 1.0, 2.0, 1.0, 2.0 ng/mL for geniposide, genipin gentiobioside, rhein, emodin, isonaringin, naringin, hesperidin and neohesperidin, respectively. The extraction recoveries of the analytes and IS from rat plasma were all more than 86.0%. The method was fully validated and applied to compare the pharmacokinetic profiles of the analytes in normal and cholestatic liver injury (CLI) rats after oral administration of ZZDHD. Results showed that there were remarkable differences in pharmacokinetic properties of the analytes between normal and CLI group.

  3. Analysis of glucose and lactate in hippocampal dialysates of rats during the operant conditioned reflex using microdialysis.

    Science.gov (United States)

    Dong, Yu; Wang, Lei; Shangguan, Dihua; Yu, Xiao; Zhao, Rui; Han, Huiwan; Liu, Guoquan

    2003-07-01

    Changes of extracellular glucose and lactate in hippocampus for freely moving rats during the operant conditioned reflex were examined simultaneously. Samples of the dialysate were assayed for both glucose and lactate using in vivo microdialysis and a microbore flow injection analysis-immobilized enzyme reactor-electrochemical detection (FIA-IMER-ECD) system. Microdialysis samplings were conducted in a Skinner box where lights were delivered as conditioned stimuli (CS) paired with foot shocks as unconditioned stimuli (US). In the treatment group the concentration of glucose and lactate showed no fluctuations during the whole process. However, in the control group in which the rats were exposed to many foot shocks, lactate levels decreased by 19% below baseline during the behavioral session and glucose showed a delayed decrease (by 18%). Compared with glucose, lactate can immediately indicate the dynamic changes in brain.

  4. Simultaneous determination and pharmacokinetic study of four phenolic acids in rat plasma using UFLC-MS/MS after intravenous administration of salvianolic acid for injection.

    Science.gov (United States)

    Xie, Xiuman; Miao, Jingzhuo; Sun, Wanyang; Huang, Jingyi; Li, Dongxiang; Li, Shuming; Tong, Ling; Sun, Guoxiang

    2017-02-05

    A simple, sensitive and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was established for simultaneous determination and pharmacokinetic study of rosmarinic acid (RA), salvianolic acid D (Sal D), lithospermic acid (LA) and salvianolic acid B (Sal B) in rat plasma after intravenous administration of salvianolic acid for injection (SAFI). Three doses of administration, containing 14, 28 and 56mg/kg, were investigated in this study. Plasma samples were pretreated using protein precipitation (PP) with pre-cooled acetonitrile. Chromatographic separation was achieved on a CORTECS™ UPLC C18 column (1.6μm, 2.1×100mm) with a mobile phase composed of 0.1% formic acid aqueous (V/V) and 0.1% formic acid acetonitrile (V/V). Analytes were detected using electrospray ionization (ESI) source in negative ionization mode and quantified in multiple reaction monitoring (MRM) mode. The validated method is stable and reliable. No significant difference of half lives (t1/2) of four analytes at three doses was observed. Area under the curve (AUC0-∞) and peak concentration (Cmax) of the four analytes demonstrated a linear increase in across the doses with the linear correlation r of each analyte at three doses were greater than 0.95. It indicated that the pharmacokinetic behavior of SAFI is positively related to dose at the range of 14-56mg/kg.

  5. Simultaneous determination of nine model compounds in permeability samples using RP-HPLC: application to prove the cassette administration principle in single pass intestinal perfusion study in rats.

    Science.gov (United States)

    Wahajuddin; Singh, Sheelendra Pratap; Raju, K S R; Nafis, Asad; Jain, Girish Kumar

    2012-01-01

    A simple, sensitive and specific reversed phase high performance liquid chromatographic (RP-HPLC) method for simultaneous determination of atenolol, paracetamol, hydrochlorothiazide, caffeine, cephalexin, metoprolol, propranolol, ketoprofen along with phenol red (a non-absorbable compound) in samples obtained from intestinal in situ single-pass perfusion studies, was developed and validated. Chromatography was carried out on RP18 column with mobile phase comprising of 10 mM phosphate buffer (pH 2.5) and methanol in gradient mode. The calibration curves were linear for all nine permeability model compounds (r² > 0.999) across the concentration range of 1.25-40 μg/ml. The coefficient of variation for intra and inter-day assay precision was between 0.04 and 3.08% and the accuracy was between 98.39 and 109.45%. Stability studies were carried out at different storage conditions and all the analytes were found to be stable. The method was successfully applied for analysing the permeability samples obtained from in situ single pass perfusion studies. The effective permeability (P(eff)) values obtained upon cassette administration were in close proximity to the permeability values obtained upon single administration of model compounds. In conclusion, the developed RP-HPLC method can be used for high throughput cassette validation of rat in situ perfusion model for intestinal permeability assessment.

  6. Ownership Structure, Financial Decisions, and Institutional Setting: An International Analysis through Simultaneous Equations

    Directory of Open Access Journals (Sweden)

    Félix J. López-Iturriaga

    2012-01-01

    Full Text Available We analyze the mutual relations among firms’ capital structure, ownership structure, and valuation. Through the estimation of a system of simultaneous equations for a sample of 1,130 firms from 16 countries from both the common law and the civil law environments, our results confirm the differential effect of ownership structure on firms’ value in each setting. Whereas in civil law firms the higher ownership concentration results in an entrenchment and an alignment effect, in the common law firms higher ownership concentration increases the value of the firm. Second, we corroborate the endogeneity of ownership structure since we find that ownership structure is affected by the value of the firm and by the capital structure. Third, our results suggest that corporate finance decisions are taken simultaneously with other mechanisms of corporate governance and conditional on firms’ valuation.

  7. Simultaneous Strain and Temperature Measurement with Optical Fiber Gratings: Error Analysis

    Institute of Scientific and Technical Information of China (English)

    JIA Hongzhi; LI Yulin

    2000-01-01

    Many schemes designed to simultaneously measure strain and temperature with optical fiber grating sensors have been reported in recent years. In this paper, the influence of systematic errors associated with the measurement process is analyzed and the error formulas are derived. The results are applied to a range of techniques that are of current interest in the literature. The performance of these schemes is contrasted with respect to the influence of wavelength measurement error and sensitivity matrix error.

  8. Simultaneous Determination of Preservatives in Dairy Products by HPLC and Chemometric Analysis

    OpenAIRE

    Fatemeh Zamani Mazdeh; Sima Sasanfar; Anita Chalipour; Elham Pirhadi; Ghazal Yahyapour; Armin Mohammadi; Akram Rostami; Mohsen Amini; Mannan Hajimahmoodi

    2017-01-01

    Cheese and yogurt are two kinds of nutritious dairy products that are used worldwide. The major preservatives in dairy products are sodium benzoate, potassium sorbate, and natamycin. The maximum permitted levels for these additives in cheese and yogurt are established according to Iranian national standards. In this study, we developed a method to detect these preservatives in dairy products by reversed phase chromatography with UV detection in 220 nm, simultaneously. This method was performe...

  9. Simultaneous derivative spectrophotometric analysis of doxylamine succinate, pyridoxine hydrochloride and folic Acid in combined dosage forms.

    Science.gov (United States)

    Pathak, A; Rajput, S J

    2008-01-01

    Two UV spectrophotometric methods have been developed, based on first derivative spectrophotometry for simultaneous estimation of doxylamine succinate, pyridoxine hydrochloride, and folic acid in tablet formulations. In method I, the concentrations of these drugs were determined by using linear regression equation. Method II is also based on first derivative spectrophotometry however simultaneous equations (Vierdot's method) were derived on derivative spectra. The first derivative amplitudes at 270.0, 332.8 and 309.2 nm were utilized for simultaneous estimation of these drugs respectively by both methods. In both the methods, linearity was obtained in the concentration range 2.5-50 mug/ml, 1-40 mug/ml and 1-30 mug/ml for doxylamine succinate, pyridoxine hydrochloride, and folic acid respectively. The developed methods show best results in terms of linearity, accuracy, precision, LOD, LOQ and ruggedness for standard laboratory mixtures of pure drugs and marketed formulations. The common excipients and additives did not interfere in their determinations.

  10. Simultaneous derivative spectrophotometric analysis of doxylamine succinate, pyridoxine hydrochloride and folic acid in combined dosage forms

    Directory of Open Access Journals (Sweden)

    Pathak A

    2008-01-01

    Full Text Available Two UV spectrophotometric methods have been developed, based on first derivative spectrophotometry for simultaneous estimation of doxylamine succinate, pyridoxine hydrochloride, and folic acid in tablet formulations. In method I, the concentrations of these drugs were determined by using linear regression equation. Method II is also based on first derivative spectrophotometry however simultaneous equations (Vierdot′s method were derived on derivative spectra. The first derivative amplitudes at 270.0, 332.8 and 309.2 nm were utilized for simultaneous estimation of these drugs respectively by both methods. In both the methods, linearity was obtained in the concentration range 2.5-50 µg/ml, 1-40 µg/ml and 1-30 µg/ml for doxylamine succinate, pyridoxine hydrochloride, and folic acid respectively. The developed methods show best results in terms of linearity, accuracy, precision, LOD, LOQ and ruggedness for standard laboratory mixtures of pure drugs and marketed formulations. The common excipients and additives did not interfere in their determinations.

  11. Simultaneous determination of Nifuroxazide and Drotaverine hydrochloride in pharmaceutical preparations by bivariate and multivariate spectral analysis

    Science.gov (United States)

    Metwally, Fadia H.

    2008-02-01

    The quantitative predictive abilities of the new and simple bivariate spectrophotometric method are compared with the results obtained by the use of multivariate calibration methods [the classical least squares (CLS), principle component regression (PCR) and partial least squares (PLS)], using the information contained in the absorption spectra of the appropriate solutions. Mixtures of the two drugs Nifuroxazide (NIF) and Drotaverine hydrochloride (DRO) were resolved by application of the bivariate method. The different chemometric approaches were applied also with previous optimization of the calibration matrix, as they are useful in simultaneous inclusion of many spectral wavelengths. The results found by application of the bivariate, CLS, PCR and PLS methods for the simultaneous determinations of mixtures of both components containing 2-12 μg ml -1 of NIF and 2-8 μg ml -1 of DRO are reported. Both approaches were satisfactorily applied to the simultaneous determination of NIF and DRO in pure form and in pharmaceutical formulation. The results were in accordance with those given by the EVA Pharma reference spectrophotometric method.

  12. Analysis of Seizure EEG in Kindled Epileptic Rats

    Directory of Open Access Journals (Sweden)

    A. K. Sen

    2007-01-01

    Full Text Available Using wavelet analysis we have detected the presence of chirps in seizure EEG signals recorded from kindled epileptic rats. Seizures were induced by electrical stimulation of the amygdala and the EEG signals recorded from the amygdala were analyzed using a continuous wavelet transform. A time–frequency representation of the wavelet power spectrum revealed that during seizure the EEG signal is characterized by a chirp-like waveform whose frequency changes with time from the onset of seizure to its completion. Similar chirp-like time–frequency profiles have been observed in newborn and adult patients undergoing epileptic seizures. The global wavelet spectrum depicting the variation of power with frequency showed two dominant frequencies with the largest amounts of power during seizure. Our results indicate that a kindling paradigm in rats can be used as an animal model of human temporal lobe epilepsy to detect seizures by identifying chirp-like time–frequency variations in the EEG signal.

  13. Proteomic analysis of indium embryotoxicity in cultured postimplantation rat embryos.

    Science.gov (United States)

    Usami, Makoto; Nakajima, Mikio; Mitsunaga, Katsuyoshi; Miyajima, Atsuko; Sunouchi, Momoko; Doi, Osamu

    2009-12-01

    Indium embryotoxicity was investigated by proteomic analysis with two-dimensional electrophoresis of rat embryos cultured from day 10.5 of gestation for 24h in the presence of 50 microM indium trichloride. In the embryo proper, indium increased quantity of several protein spots including those identified as serum albumin, phosphorylated cofilin 1, phosphorylated destrin and tyrosyl-tRNA synthetase. The increased serum albumin, derived from the culture medium composed of rat serum, may decrease the toxicity of indium. The increase of phosphorylated cofilin 1 might be involved in dysmorphogenicity of indium through perturbation of actin functions. In the yolk sac membrane, indium induced quantitative and qualitative changes in the protein spots. Proteins from appeared spots included stress proteins, and those from decreased or disappeared spots included serum proteins, glycolytic pathway enzymes and cytoskeletal proteins, indicating yolk sac dysfunction. Thus, several candidate proteins that might be involved in indium embryotoxicity were identified.

  14. Simultaneous determination of ten Aconitum alkaloids in rat tissues by UHPLC-MS/MS and its application to a tissue distribution study on the compatibility of Heishunpian and Fritillariae thunbergii Bulbus.

    Science.gov (United States)

    Yang, Bin; Xu, Yanyan; Wu, Yuanyuan; Wu, Huanyu; Wang, Yuan; Yuan, Lei; Xie, Jiabin; Li, Yubo; Zhang, Yanjun

    2016-10-15

    A rapid, sensitive and selective ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for simultaneous determination of ten Aconitum alkaloids in rat tissues. The tissue samples were prepared by a simple procedure protein precipitation with acetonitrile containing 0.1% acetic acid and separated on an Agilent XDB C18 column (4.6 mm×50mm, 1.8μm) using gradient elution with a mobile phase consisting of water and acetonitrile (both containing 0.1% formic acid) at a flow rate of 0.3mL/min. The quantitive determination was performed on an electrospray ionization (ESI) triple quadrupole tandem mass spectrometer using selective reaction monitoring (SRM) under positive ionization mode. The established method was fully validated according to the USA Food and Drug Administration (FDA) bioanalytical method validation guidance and the results demonstrated that the method was sensitive and selective with the lowest limits of quantification (LLOQ) at 0.025ng/mL in rat tissue homogenates. Meanwhile, the linearity, precision, accuracy, extraction recovery, matrix effect and stability were all within the required limits of biological sample analysis. After method validation, the validated method was successfully applied to the tissue distribution study on the compatibility of Heishunpian (HSP, the processed product of Aconitum carmichaelii Debx) and Fritillariae thunbergii Bulbus (Zhebeimu, ZBM). The results indicated that the distribution feature of monoester diterpenoid aconitines (MDAs), diester diterpenoid aconitines (DDAs) and non-ester alkaloids (NEAs) were inconsistency, and the compatibility of HSP and ZBM resulted in the distribution amount of DDAs increased in tissues. What's more, the results could provide the reliable basis for systematic research on the substance foundation of the compatibility of the herbal pair.

  15. The simultaneous loop flow correction analysis in the water feed network of Minkok Edjombo (SOUTH –CAMEROON.

    Directory of Open Access Journals (Sweden)

    MENDOMO MEYE SERGES

    2014-11-01

    Full Text Available With increasing population growth and industrial development, water flow rates and other hydraulic requirements associated with water distribution systems have been estimated to increase both national and local scale. Water shortage will cause inconvenience to people’s life and it will impact city function and industrial production. Hence to overcome this problem design and analysis of water distribution system is necessary to get optimal discharge. In this paper a water pipeline network analysis with a case study of a small city (MinkokEdjombo in the southern Cameroon system has been undertaken. What prompted this study is that the case study has a lot of fluctuations in its head loss. Also, the discharge is not proportional to the pipe diameter. The study therefore adopted simultaneous loop flow correction method because it computes simultaneous flows corrections for all loops, hence, the best since computational procedures takes into account the iterative influence of flow corrections between loops which have common pipes. After applying the simultaneous loop flow correction analyze in a twenty-four sampled pipeline network, a drastic reduction in head loss and regular line along the axis was observed. Besides, the rate at which the water flows was observed to be proportional to the pipe diameter. Hence, the method is a useful aid in planning, designing and operating of reticulated pipeline network for higher efficiency and improved economy.

  16. Three-dimensional finite element analysis of stress distribution in inlay-restored mandibular first molar under simultaneous thermomechanical loads.

    Science.gov (United States)

    Çelik Köycü, Berrak; Imirzalioğlu, Pervin; Özden, Utku Ahmet

    2016-01-01

    Functional occlusal loads and intraoral temperature changes create stress in teeth. The purpose of this study was to evaluate the impact of simultaneous thermomechanical loads on stress distribution related to inlay restored teeth by three-dimensional finite element analysis. A mandibular first molar was constructed with tooth structures, surrounding bone and inlays of Type II gold alloy, ceramic, and composite resin. Stress patterns on the restorative materials, adhesive resin, enamel and dentin were analyzed after simulated temperature changes from 36°C to 4 or 60°C for 2 s with 200-N oblique loading. The results showed that the three types of inlays had similar stress distribution in the tooth structures and restorative materials. Concerning the adhesive resin, the composite resin inlay model exhibited lower stresses than ceramic and gold alloy inlays. Simultaneous thermomechanical loads caused high stress patterns in inlay-restored teeth. Composite resin inlays may be the better choice to avoid adhesive failure.

  17. No adverse effects detected for simultaneous whole-body exposure to multiple-frequency radiofrequency electromagnetic fields for rats in the intrauterine and pre- and post-weaning periods.

    Science.gov (United States)

    Shirai, Tomoyuki; Wang, Jianqing; Kawabe, Mayumi; Wake, Kanako; Watanabe, So-Ichi; Takahashi, Satoru; Fujiwara, Osamu

    2017-01-01

    In everyday life, people are exposed to radiofrequency (RF) electromagnetic fields (EMFs) with multiple frequencies. To evaluate the possible adverse effects of multifrequency RF EMFs, we performed an experiment in which pregnant rats and their delivered offspring were simultaneously exposed to eight different communication signal EMFs (two of 800 MHz band, two of 2 GHz band, one of 2.4 GHz band, two of 2.5 GHz band and one of 5.2 GHz band). Thirty six pregnant Sprague-Dawley (SD) 10-week-old rats were divided into three groups of 12 rats: one control (sham exposure) group and two experimental (low- and high-level RF EMF exposure) groups. The whole body of the mother rats was exposed to the RF EMFs for 20 h per day from Gestational Day 7 to weaning, and F1 offspring rats (46-48 F1 pups per group) were then exposed up to 6 weeks of age also for 20 h per day. The parameters evaluated included the growth, gestational condition and organ weights of the dams; the survival rates, development, growth, physical and functional development, memory function, and reproductive ability of the F1 offspring; and the embryotoxicity and teratogenicity in the F2 rats. No abnormal findings were observed in the dams or F1 offspring exposed to the RF EMFs or to the F2 offspring for any of the parameters evaluated. Thus, under the conditions of the present experiment, simultaneous whole-body exposure to eight different communication signal EMFs at frequencies between 800 MHz and 5.2 GHz did not show any adverse effects on pregnancy or on the development of rats.

  18. No adverse effects detected for simultaneous whole-body exposure to multiple-frequency radiofrequency electromagnetic fields for rats in the intrauterine and pre- and post-weaning periods

    Science.gov (United States)

    Shirai, Tomoyuki; Wang, Jianqing; Kawabe, Mayumi; Wake, Kanako; Watanabe, So-ichi; Takahashi, Satoru; Fujiwara, Osamu

    2017-01-01

    In everyday life, people are exposed to radiofrequency (RF) electromagnetic fields (EMFs) with multiple frequencies. To evaluate the possible adverse effects of multifrequency RF EMFs, we performed an experiment in which pregnant rats and their delivered offspring were simultaneously exposed to eight different communication signal EMFs (two of 800 MHz band, two of 2 GHz band, one of 2.4 GHz band, two of 2.5 GHz band and one of 5.2 GHz band). Thirty six pregnant Sprague-Dawley (SD) 10-week-old rats were divided into three groups of 12 rats: one control (sham exposure) group and two experimental (low- and high-level RF EMF exposure) groups. The whole body of the mother rats was exposed to the RF EMFs for 20 h per day from Gestational Day 7 to weaning, and F1 offspring rats (46–48 F1 pups per group) were then exposed up to 6 weeks of age also for 20 h per day. The parameters evaluated included the growth, gestational condition and organ weights of the dams; the survival rates, development, growth, physical and functional development, memory function, and reproductive ability of the F1 offspring; and the embryotoxicity and teratogenicity in the F2 rats. No abnormal findings were observed in the dams or F1 offspring exposed to the RF EMFs or to the F2 offspring for any of the parameters evaluated. Thus, under the conditions of the present experiment, simultaneous whole-body exposure to eight different communication signal EMFs at frequencies between 800 MHz and 5.2 GHz did not show any adverse effects on pregnancy or on the development of rats. PMID:27694283

  19. Multivariate image analysis-thin layer chromatography (MIA-TLC) for simultaneous determination of co-eluting components.

    Science.gov (United States)

    Hemmateenejad, Bahram; Mobaraki, Nabiollah; Shakerizadeh-Shirazi, Fatemeh; Miri, Ramin

    2010-07-01

    This paper addresses the solution of peak overlapping, as a fundamental problem in TLC, by multivariate analysis of the images recorded by a digital camera. We report the results of our study on the application of multivariate image analysis (MIA) for simultaneous determination of several species on thin layer chromatography (TLC) sheet for the first time. An imaging system, composed of a dark cabinet, a digital camera and a multivariate image analysis program, was prepared for recording the images of TLC plates after development of a multi-component solution. The written program was able to produce 2- and 3-dimensional chromatograms of the solutions, which were subsequently used as inputs of partial least squares, as an efficient multivariate calibration method. The ability of the proposed MIA-TLC method for simultaneous determination of the co-eluting components was validated by analysis of ternary synthetic mixtures of indicators of highly overlapped chromatograms (i.e., methyl yellow, bromocresol green and creseol red) and a real mixture of nifedipine and its photo-degradation product. By application of different strategies like principal component analysis and variable selection, models were obtained that could estimate the concentration of indicators in the external prediction set with relative errors of lower than 10% and in most cases lower than 5%.

  20. Simultaneous determination of nine lignans from Schisandra chinensis extract using ultra-performance liquid chromatography with tandem mass spectrometry in rat plasma, urine, and gastrointestinal tract samples: application to the pharmacokinetic study of Schisandra chinensis.

    Science.gov (United States)

    Kim, You-Jin; Lee, Hee Ju; Kim, Chul Young; Han, Sun-Young; Chin, Young-Won; Choi, Young Hee

    2014-10-01

    The fruit of Schisandra chinensis is a well-known herbal medicine and dietary supplement due to a variety of biological activities including antihepatotoxic and antihyperlipidemic activities. However, the simultaneous validation methodology and pharmacokinetic investigation of nine lignans of S. chinensis extract in biological samples have not been proved yet. Thus, the present study was undertaken to develop the proper sample preparation method and simultaneous analytical method of schisandrol A, gomisin J, schisandrol B, tigloylgomisin H, angeloylgomisin H, schisandrin A, schisandrin B, gomisin N, and schisandrin C in the hexane-soluble extract of S. chinensis to apply for the pharmacokinetic study in rats. All intra- and interprecisions of nine lignans were below 13.7% and accuracies were 85.1-115% and it is enough to evaluate the pharmacokinetic parameters after both intravenous and oral administration of hexane-soluble extract of S. chinensis to rats.

  1. Liquid-phase microextraction for simultaneous chromatographic analysis of three antidepressant drugs in plasma

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Dobrovolskni Porto

    2012-01-01

    Full Text Available A method using Liquid Phase Microextraction for simultaneous detection of citalopram (CIT, paroxetine (PAR and fluoxetine (FLU, using venlafaxine as internal standard, in plasma by high performance liquid chromatography with fluorescence detection was developed. The linearity was evaluated between 5.0 and 500 ng mL-1 (r > 0.99 and the limit of quantification was 2.0, 3.0 and 5.0 ng mL-1 for CIT, PAR and FLU, respectively. Therefore, it can be applied to therapeutic drug monitoring, pharmacokinetics or bioavailability studies and its advantages are that it necessary relatively inexpensive equipment and sample preparation techniques.

  2. Simultaneous analysis of the glow curves of thermoluminescence and thermally stimulated exo-electron emission

    CERN Document Server

    Sakurai, T; Fukuda, Y

    1999-01-01

    A new method to analyse the glow curves of thermoluminescence (TL) and thermally stimulated exo-electron emission (TSEE) simultaneously is proposed. The method is based on a model consisting of one trap-one recombination centre for TL and thermionic emission for TSEE. A set of coupled differential equations, describing the charge flow, is numerically solved without any approximation, using the integrated experimental glow curves, maximum conditions and boundary conditions of TL and TSEE. The computer simulation is carried out in the two following cases: the case of correlation between TL and TSEE; and the case of no correlation between TL and TSEE. (author)

  3. Comparative secretome analysis of rat stomach under different nutritional status

    Directory of Open Access Journals (Sweden)

    Lucia L. Senin

    2015-06-01

    Full Text Available The fact that gastric surgery is at the moment the most effective treatment to fight against obesity highlights the relevance of gastric derived proteins as potential targets to treat this pathology. Taking advantage of a previously established gastric explant model for endocrine studies, the proteomic analysis of gastric secretome was performed. To validate this gastric explant system for proteomic analysis, the identification of ghrelin, a classical gastric derived peptide, was performed by MS. In addition, the differential analysis of gastric secretomes under differential nutritional status (control feeding vs fasting vs re-feeding was performed. The MS identified proteins are showed in the present manuscript. The data supplied in this article is related to the research article entitled “Comparative secretome analysis of rat stomach under different nutritional status” [1].

  4. Data from quantitative label free proteomics analysis of rat spleen

    Directory of Open Access Journals (Sweden)

    Khadar Dudekula

    2016-09-01

    Full Text Available The dataset presented in this work has been obtained using a label-free quantitative proteomic analysis of rat spleen. A robust method for extraction of proteins from rat spleen tissue and LC-MS-MS analysis was developed using a urea and SDS-based buffer. Different fractionation methods were compared. A total of 3484 different proteins were identified from the pool of all experiments run in this study (a total of 2460 proteins with at least two peptides. A total of 1822 proteins were identified from nine non-fractionated pulse gels, 2288 proteins and 2864 proteins were identified by SDS-PAGE fractionation into three and five fractions respectively. The proteomics data are deposited in ProteomeXchange Consortium via PRIDE PXD003520, Progenesis and Maxquant output are presented in the supported information. The generated list of proteins under different regimes of fractionation allow assessing the nature of the identified proteins; variability in the quantitative analysis associated with the different sampling strategy and allow defining a proper number of replicates for future quantitative analysis.

  5. Data from quantitative label free proteomics analysis of rat spleen.

    Science.gov (United States)

    Dudekula, Khadar; Le Bihan, Thierry

    2016-09-01

    The dataset presented in this work has been obtained using a label-free quantitative proteomic analysis of rat spleen. A robust method for extraction of proteins from rat spleen tissue and LC-MS-MS analysis was developed using a urea and SDS-based buffer. Different fractionation methods were compared. A total of 3484 different proteins were identified from the pool of all experiments run in this study (a total of 2460 proteins with at least two peptides). A total of 1822 proteins were identified from nine non-fractionated pulse gels, 2288 proteins and 2864 proteins were identified by SDS-PAGE fractionation into three and five fractions respectively. The proteomics data are deposited in ProteomeXchange Consortium via PRIDE PXD003520, Progenesis and Maxquant output are presented in the supported information. The generated list of proteins under different regimes of fractionation allow assessing the nature of the identified proteins; variability in the quantitative analysis associated with the different sampling strategy and allow defining a proper number of replicates for future quantitative analysis.

  6. Peptidomic Analysis of Rat Plasma: Proteolysis in Hemorrhagic Shock.

    Science.gov (United States)

    Aletti, Federico; Maffioli, Elisa; Negri, Armando; Santamaria, Marco H; DeLano, Frank A; Kistler, Erik B; Schmid-Schönbein, Geert W; Tedeschi, Gabriella

    2016-05-01

    It has been previously shown that intestinal proteases translocate into the circulation during hemorrhagic shock and contribute to proteolysis in distal organs. However, consequences of this phenomenon have not previously been investigated using high-throughput approaches. Here, a shotgun label-free quantitative proteomic approach was utilized to compare the peptidome of plasma samples from healthy and hemorrhagic shock rats to verify the possible role of uncontrolled proteolytic activity in shock. Plasma was collected from rats after hemorrhagic shock (HS) consisting of 2-h hypovolemia followed by 2-h reperfusion, and from healthy control (CTRL) rats. A new two-step enrichment method was applied to selectively extract peptides and low molecular weight proteins from plasma, and directly analyze these samples by tandem mass spectrometry. One hundred twenty-six circulating peptides were identified in CTRL and 295 in HS animals. Ninety-six peptides were present in both conditions; of these, 57 increased and 30 decreased in shock. In total, 256 peptides were increased or present only in HS confirming a general increase in proteolytic activity in shock. Analysis of the proteases that potentially generated the identified peptides suggests that the larger relative contribution to the proteolytic activity in shock is due to chymotryptic-like proteases. These results provide quantitative confirmation that extensive, system-wide proteolysis is part of the complex pathologic phenomena occurring in hemorrhagic shock.

  7. Simultaneous determination of timosaponin B-II and A-III in rat plasma by LC-MS/MS and its application to pharmacokinetic study.

    Science.gov (United States)

    Feng, Yi; Chen, Baoting; Lin, Aihua; Liu, Yiming

    2014-08-15

    A rapid, specific and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the simultaneous determination of timosaponin B-II (TB-II) and A-III (TA-III) in rat plasma. Plasma samples were pretreated via simple protein precipitation with acetonitrile and ginsenoside Rg2 was used as internal standard. Chromatographic separation was carried out on an Agilent XDB-C8 (150 mm × 2.1mm i.d., 5 μm) column by isocratic elution with acetonitrile-2 mmol/L ammonium acetate (55:45, v/v). The detection was performed on a Sciex API 4000(+) triple-quadrupole tandem mass spectrometer with TurboIonSpray ionization (ESI) inlet via the negative ion multiple reaction monitoring (MRM) mode. The results showed that the calibration curve was linear in the concentration range of 3-3,000 ng/mL for TB-II and 0.3-3,000 ng/mL for TA-III, respectively. The intra- and inter-day precisions were less than 13.25%, and the accuracy ranged from 100.88% to 104.07% at three QC levels for both. The pharmacokinetic profiles of TB-II and TA-III in timosaponins (total timosaponin) at three dose levels (TB-II 150, 300, 600 mg/kg and TA-III 0.59, 1.17, 2.34 mg/kg, respectively) and in timosaponins-Huangbai alkaloids mixtures (1:1, 1:3, w/w, TB-II 300 mg/kg and TA-III 1.17 mg/kg) were studied for the first time in rats by this LC-MS/MS method. After single oral administration of timosaponins, mean Cmax and AUC0-t of TB-II and TA-III increased but non-proportional to the oral doses. When timosaponins-Huangbai alkaloids (1:1, 1:3, w/w) mixtures were administered, Cmax and AUC0-t of TB-II in the mixtures were obviously higher than the corresponding values in timosaponins at the same dose level. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Changes in testosterone and dihydrotestosterone levels in male rat accessory sex organs, serum, and seminal fluid after castration: establishment of a new highly sensitive simultaneous androgen measurement method.

    Science.gov (United States)

    Kashiwagi, Bunzo; Shibata, Yasuhiro; Ono, Yoshihiro; Suzuki, Ryota; Honma, Seijiro; Suzuki, Kazuhiro

    2005-01-01

    It is known that abnormal androgen dynamics in the tissues is a cause of androgen-dependent disorders. Investigation of tissue androgen levels could provide a clue to the elucidation of disorders. However, it is difficult to measure a trace amount of androgen in the tissues. We established a highly sensitive simultaneous quantification method of testosterone and dihydrotestosterone (DHT), which play the most important roles in the body among androgenic steroids in trace amounts, and investigated time course changes in testosterone and DHT levels in male accessory sex organs, serum, and seminal fluid after castration in rat models. In addition, changes in the testosterone/DHT ratio of male accessory sex organs and seminal fluid were observed. The simultaneous testosterone and DHT measurement method established by us was validated. Intra-assay variation and interassay precision and accuracy were all within +/-20%, and the quantification limits of testosterone and DHT were both 15.6 pg/g. With the use of this method, the testosterone and DHT levels in the prostate, seminal vesicles, and serum immediately after castration were similar to those previously reported. The testosterone and DHT levels were 350 pg/g and 605 pg/g, respectively; which showed dominance of DHT in seminal fluid, although it was not as marked as that in the male accessory sex organs. Androgens decreased with time after castration in the accessory sex organs, serum, and seminal fluid. In the prostate and seminal vesicles, testosterone and DHT decreased to about 50% and about 2% of the normal levels, respectively, 72 hours after castration. The serum levels were under the quantification limits 6 hours after castration and thereafter. In seminal fluid, the testosterone and DHT levels decreased to 49% and 35% of normal levels, respectively, 72 hours after castration. The testosterone/DHT ratio in the male accessory sex organs was lower in the prostate (0.06) than in the seminal vesicles (0

  9. Analysis of lightning-ionosphere interaction using simultaneous records of source current and 380 km distant electric field

    Science.gov (United States)

    Azadifar, Mohammad; Li, Dongshuai; Rachidi, Farhad; Rubinstein, Marcos; Diendorfer, Gerhard; Schulz, Wolfgang; Pichler, Hannes; Rakov, Vladimir A.; Paolone, Mario; Pavanello, Davide

    2017-06-01

    We present simultaneous current and wideband electric field waveforms at 380 km associated with upward lightning flashes initiated from the Säntis Tower, Switzerland. To the best of our knowledge, the dataset presented in this study includes the first simultaneous records of lightning currents and associated fields featuring ionospheric reflections for natural upward flashes, and the longest distance at which natural upward lightning fields have been measured simultaneously with their causative currents. The intervals between the groundwave and skywave arrival times are used to estimate ionospheric reflection heights during day and night times using the so-called zero-to-zero and peak-to-peak methods. During daytime, the mean ionospheric reflection heights, obtained using the two different delay estimation approaches, are about 78 and 76 km, corresponding to the D layer. The mean reflection height at nighttime is about 90 and 89 km, corresponding to the E layer. We present a full-wave, finite-difference time-domain (FDTD) analysis of the electric field propagation including the effect of the ionospheric reflections. The FDTD simulation results are compared with the measured fields associated with upward flashes initiated from the Säntis Tower. It is found that the model reproduces reasonably well the measured waveforms and the times of arrival of the one-hop and two-hop skywaves relative to the groundwave.

  10. Simultaneous determination of vanadium(IV) and vanadium(V) by flow injection analysis using kinetic spectrophotometry with Xylenol Orange.

    Science.gov (United States)

    Oguma, Koichi; Yoshioka, Osamu; Noro, Junji; Sakurai, Hiroki

    2012-07-15

    A flow injection method of analysis has been developed for the simultaneous determination of V(IV) and V(V) using the difference in the rate of complex formation between vanadium in two different oxidation states and Xylenol Orange in acidic media. The proposed method used a spectrophotometric detector equipped with two flow cells aligned with the same optical path to yield two successive peaks per each sample injection. V(IV) and V(V) were determined by solving simultaneous equations in two unknowns, the concentrations of V(IV) and V(V), obtained from measurements of the two peak heights. The detection limit calculated as 3σ of noise signals was 0.01μg/mL for both V(IV) and V(V). The relative standard deviations for V(IV) and V(V) at the 1μg/mL level were 1.6 and 2.4%, respectively. The sample throughput was found to be about 19h(-1). The proposed method was successfully applied to the simultaneous determination of V(IV) and V(V) in synthetic scrubbing solutions used in desulphurization with reasonable accuracy.

  11. Rats

    Directory of Open Access Journals (Sweden)

    Alexey Kondrashov

    2012-01-01

    Full Text Available We aimed to perform a chemical analysis of both Alibernet red wine and an alcohol-free Alibernet red wine extract (AWE and to investigate the effects of AWE on nitric oxide and reactive oxygen species production as well as blood pressure development in normotensive Wistar Kyoto (WKY and spontaneously hypertensive rats (SHRs. Total antioxidant capacity together with total phenolic and selected mineral content was measured in wine and AWE. Young 6-week-old male WKY and SHR were treated with AWE (24,2 mg/kg/day for 3 weeks. Total NOS and SOD activities, eNOS and SOD1 protein expressions, and superoxide production were determined in the tissues. Both antioxidant capacity and phenolic content were significantly higher in AWE compared to wine. The AWE increased NOS activity in the left ventricle, aorta, and kidney of SHR, while it did not change NOS activity in WKY rats. Similarly, increased SOD activity in the plasma and left ventricle was observed in SHR only. There were no changes in eNOS and SOD1 expressions. In conclusion, phenolics and minerals included in AWE may contribute directly to increased NOS and SOD activities of SHR. Nevertheless, 3 weeks of AWE treatment failed to affect blood pressure of SHR.

  12. Simultaneous Determination of Bergapten, Imperatorin, Notopterol, and Isoimperatorin in Rat Plasma by High Performance Liquid Chromatography with Fluorescence Detection and Its Application to Pharmacokinetic and Excretion Study after Oral Administration of Notopterygium incisum Extract

    Directory of Open Access Journals (Sweden)

    John Teye Azietaku

    2016-01-01

    Full Text Available A specific, sensitive, and reliable high performance liquid chromatography with fluorescence detection (HPLC-FLD was first optimized and then used in the simultaneous quantification of bergapten, imperatorin, notopterol, and isoimperatorin in rat plasma using osthole as the internal standard. Liquid-liquid extraction with ethyl acetate was employed in treating the rat plasma samples obtained. Separation was carried out with a Hedera™ ODS column (4.6 × 250 mm, 5 μm by gradient elution at a temperature of 40°C. Excitation and emission of the fluorescence detector were set to 300 and 490 nm, respectively. The lower limits of quantification for bergapten, imperatorin, notopterol, and isoimperatorin in rat plasma were 4, 40, 4, and 2 ng mL−1, respectively. The intraday and interday precision and accuracy for the four coumarins were within acceptable criteria. The recovery of the method was satisfactory with a range of 80.3–114%. The validated method was successfully used for the simultaneous determination of the four coumarins in Notopterygium incisum extracts and also for the pharmacokinetic and excretion study of bergapten, imperatorin, notopterol, and isoimperatorin in rats.

  13. Simultaneous Determination of Bergapten, Imperatorin, Notopterol, and Isoimperatorin in Rat Plasma by High Performance Liquid Chromatography with Fluorescence Detection and Its Application to Pharmacokinetic and Excretion Study after Oral Administration of Notopterygium incisum Extract

    Science.gov (United States)

    Teye Azietaku, John; Yu, Xie-an; Li, Jin; Hao, Jia; An, Mingrui; Tan, Zhijing

    2016-01-01

    A specific, sensitive, and reliable high performance liquid chromatography with fluorescence detection (HPLC-FLD) was first optimized and then used in the simultaneous quantification of bergapten, imperatorin, notopterol, and isoimperatorin in rat plasma using osthole as the internal standard. Liquid-liquid extraction with ethyl acetate was employed in treating the rat plasma samples obtained. Separation was carried out with a Hedera™ ODS column (4.6 × 250 mm, 5 μm) by gradient elution at a temperature of 40°C. Excitation and emission of the fluorescence detector were set to 300 and 490 nm, respectively. The lower limits of quantification for bergapten, imperatorin, notopterol, and isoimperatorin in rat plasma were 4, 40, 4, and 2 ng mL−1, respectively. The intraday and interday precision and accuracy for the four coumarins were within acceptable criteria. The recovery of the method was satisfactory with a range of 80.3–114%. The validated method was successfully used for the simultaneous determination of the four coumarins in Notopterygium incisum extracts and also for the pharmacokinetic and excretion study of bergapten, imperatorin, notopterol, and isoimperatorin in rats. PMID:28115935

  14. Effects of simultaneous dietary fish oil ingestion and sulfur amino acid supplementation on the lipid metabolism in hepatoma-bearing rats with hyperlipidemia.

    Science.gov (United States)

    Kawasaki, Masashi; Miura, Yutaka; Funabiki, Ryuhei; Yagasaki, Kazumi

    2010-01-01

    The effects of simultaneous dietary fish oil ingestion and sulfur amino acid (L-methionine and L-cystine) supplementation on serum lipid concentrations and various parameters related to the lipid metabolism were studied in Donryu rats subcutaneously implanted with an ascites hepatoma cell line, AH109A. A diet containing 10% fish oil was found to reduce serum triglyceride, total cholesterol, (very-low-density lipoprotein plus low-density lipoprotein)-cholesterol, phospholipid and nonesterified fatty acid (NEFA) concentrations in these animals, and dietary supplementation of 1.2% L-methionine and L-cystine also suppressed these serum lipid concentrations. Hepatic fatty acid synthesis and the availability of serum NEFA were decreased, and epididymal adipose tissue lipoprotein lipase (LPL) activity was elevated by dietary fish oil, while LPL activity in various tissues and hepatic fatty acid oxidation were increased by dietary sulfur amino acids, resulting in a reduction in the serum triglyceride concentration by dietary fish oil and sulfur amino acids, respectively. Dietary fish oil suppressed the hepatoma-induced increase in cholesterogenesis in the host liver, and dietary methionine and cystine enhanced bile acid excretion into feces, which were the causes of the hypocholesterolemic effect. In these serum lipid concentrations, there were significant effects of fish oil ingestion and sulfur amino acid supplementation, but no significant interaction between these two factors was seen. These results indicate that dietary fish oil and sulfur amino acid, L-methionine and L-cystine, have hypolipidemic effects in cancer-related hyperlipidemia, and that the effects of these two factors on the decrease in these serum lipid concentrations are additive; these two factors may affect the lipid metabolism via different pathways and mechanisms.

  15. Simultaneous alterations of brain and plasma serotonin concentrations and liver cytochrome P450 in rats fed on a tryptophan-free diet.

    Science.gov (United States)

    Kot, Marta; Pilc, Andrzej; Daniel, Władysława A

    2012-10-01

    Our previous study suggested involvement of the brain serotonergic system in the regulation of liver cytochrome P450 (CYP). The aim of the present study was to demonstrate simultaneous responsiveness of liver CYP and the peripheral and brain serotonergic systems to a tryptophan deficient diet during three days and one or three weeks of ingestion. The concentrations of serotonin, noradrenaline, dopamine and their metabolites were measured in blood plasma, the hypothalamus and brain stem of male rats. The enzyme activity and protein levels in the liver were determined for isoforms CYP1A, CYP2A, CYP2B, CYP2C6, CYP2C11, CYP2D and CYP3A. A three-day tryptophan-free diet increased serotonin content in the hypothalamus (but not in the brain stem or plasma). After one week, the level of serotonin was not changed in the brain, but was markedly increased in the plasma. A three week tryptophan restriction significantly reduced the concentration of serotonin in the brain and plasma. Changes in CYP2C6 and CYP2C11 (an increase and a decrease, respectively) were maintained throughout the experiment, while those found in other CYP isoforms varied, which usually resulted in a gradual increase in the enzyme activity within three weeks. The observed alterations in liver CYPs suggest involvement of both central and peripheral serotonin in the regulation of liver CYP expression whose mechanism is discussed. In conclusion, a deficit in tryptophan in the diet may be responsible for very serious food-cytochrome P450 and food-drug metabolism interactions. Interactions of this type may also refer to drugs acting via serotonergic system.

  16. Simultaneous UFLC-ESI-MS/MS determination of piperine and piperlonguminine in rat plasma after oral administration of alkaloids from Piper longum L.: application to pharmacokinetic studies in rats.

    Science.gov (United States)

    Liu, Junhui; Bi, Ying; Luo, Rong; Wu, Xia

    2011-10-01

    The alkaloids from Piper longum L. showed protective effects on Parkinson's disease models in our previous study and piperine and piperlonguminine were the two main constituents in the alkaloids. The present study aimed at developing a rapid, sensitive, and accurate UFLC-ESI-MS/MS method and validating it for the simultaneous determination of piperine and piperlonguminine in rat plasma using terfenadine as the internal standard. The analytes and internal standard (IS) were extracted from rat plasma using a simple protein precipitation by adding methanol/acetonitrile (1:1, v/v). A Phenomenex Gemini 3 u C18 column (20 mm × 2.00 mm, 3 μm) was used to separate the analytes and IS using a gradient mode system with a mobile phase consisting of water with 0.1% formic acid (mobile phase A) and acetonitrile with 0.1% formic acid (mobile phase B) at a flow rate of 0.4 mL/min and an operating column temperature of 25°C. The total analytical run time was 4 min. The detection was performed using the positive ion electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode with transitions at m/z 286.1-201.1 for piperine, m/z 274.0-201.1 for piperlonguminine, and m/z 472.4-436.4 for the IS. The calibration curves were both linear (r>0.995) over a concentration range of 1.0 to 1000 ng/mL; the lower limit of quantification (LLOQ) was 1.0 ng/mL for both piperine and piperlonguminine. The intra-day and inter-day precisions (RSD %) were alkaloids from P. longum L.

  17. PCF-based cavity enhanced spectroscopic sensors for simultaneous multicomponent trace gas analysis.

    Science.gov (United States)

    Nakaema, Walter M; Hao, Zuo-Qiang; Rohwetter, Philipp; Wöste, Ludger; Stelmaszczyk, Kamil

    2011-01-01

    A multiwavelength, multicomponent CRDS gas sensor operating on the basis of a compact photonic crystal fibre supercontinuum light source has been constructed. It features a simple design encompassing one radiation source, one cavity and one detection unit (a spectrograph with a fitted ICCD camera) that are common for all wavelengths. Multicomponent detection capability of the device is demonstrated by simultaneous measurements of the absorption spectra of molecular oxygen (spin-forbidden b-X branch) and water vapor (polyads 4v, 4v + δ) in ambient atmospheric air. Issues related to multimodal cavity excitation, as well as to obtaining the best signal-to-noise ratio are discussed together with methods for their practical resolution based on operating the cavity in a "quasi continuum" mode and setting long camera gate widths, respectively. A comprehensive review of multiwavelength CRDS techniques is also given.

  18. Bayesian simultaneous equation models for the analysis of energy intake and partitioning in growing pigs

    DEFF Research Database (Denmark)

    Strathe, Anders Bjerring; Jørgensen, Henry; Kebreab, E

    2012-01-01

    ABSTRACT SUMMARY The objective of the current study was to develop Bayesian simultaneous equation models for modelling energy intake and partitioning in growing pigs. A key feature of the Bayesian approach is that parameters are assigned prior distributions, which may reflect the current state...... of nature. In the models, rates of metabolizable energy (ME) intake, protein deposition (PD) and lipid deposition (LD) were treated as dependent variables accounting for residuals being correlated. Two complementary equation systems were used to model ME intake (MEI), PD and LD. Informative priors were...... genders (barrows, boars and gilts) selected on the basis of similar birth weight. The pigs were fed four diets based on barley, wheat and soybean meal supplemented with crystalline amino acids to meet or exceed Danish nutrient requirement standards. Nutrient balances and gas exchanges were measured at c...

  19. PCF-Based Cavity Enhanced Spectroscopic Sensors for Simultaneous Multicomponent Trace Gas Analysis

    Directory of Open Access Journals (Sweden)

    Kamil Stelmaszczyk

    2011-01-01

    Full Text Available A multiwavelength, multicomponent CRDS gas sensor operating on the basis of a compact photonic crystal fibre supercontinuum light source has been constructed. It features a simple design encompassing one radiation source, one cavity and one detection unit (a spectrograph with a fitted ICCD camera that are common for all wavelengths. Multicomponent detection capability of the device is demonstrated by simultaneous measurements of the absorption spectra of molecular oxygen (spin-forbidden b-X branch and water vapor (polyads 4v, 4v + d in ambient atmospheric air. Issues related to multimodal cavity excitation, as well as to obtaining the best signal-to-noise ratio are discussed together with methods for their practical resolution based on operating the cavity in a “quasi continuum” mode and setting long camera gate widths, respectively. A comprehensive review of multiwavelength CRDS techniques is also given.

  20. Simultaneous measurements of anisotropic thermal diffusivity and thermal effusivity of liquid crystals using temperature wave analysis method

    Science.gov (United States)

    Ryu, Meguya; Morikawa, Junko

    2016-11-01

    Simultaneous measurements of anisotropic thermal diffusivity and thermal effusivity in the phase transition of the liquid crystal 4‧-(pentyloxy)-4-biphenyl carbonitrile (5OCB), Cry-53-N-68-Iso, are demonstrated using the temperature wave analysis method. A detailed calibration procedure for determining thermal diffusivity (α), thermal effusivity (e), thermal conductivity (λ), and heat capacity per unit volume (Cp ρ) is proposed utilizing hexaethylene glycol monododecyl ether (C12E6) as a calibration material, the heat capacity of which is precisely determined using a calorimetric method.

  1. Simultaneous achiral-chiral analysis of pharmaceutical compounds using two-dimensional reversed phase liquid chromatography-supercritical fluid chromatography.

    Science.gov (United States)

    Venkatramani, C J; Al-Sayah, Mohammad; Li, Guannan; Goel, Meenakshi; Girotti, James; Zang, Lisa; Wigman, Larry; Yehl, Peter; Chetwyn, Nik

    2016-02-01

    A new interface was designed to enable the coupling of reversed phase liquid chromatography (RPLC) and supercritical fluid chromatography (SFC). This online two-dimensional chromatographic system utilizing RPLC in the first dimension and SFC in the second was developed to achieve simultaneous achiral and chiral analysis of pharmaceutical compounds. The interface consists of an eight-port, dual-position switching valve with small volume C-18 trapping columns. The peaks of interest eluting from the first RPLC dimension column were effectively focused as sharp concentration pulses on small volume C-18 trapping column/s and then injected onto the second dimension SFC column. The first dimension RPLC separation provides the achiral purity result, and the second dimension SFC separation provides the chiral purity result (enantiomeric excess). The results are quantitative enabling simultaneous achiral, chiral analysis of compounds. The interface design and proof of concept demonstration are presented. Additionally, comparative studies to conventional SFC and case studies of the applications of 2D LC-SFC in pharmaceutical analysis is presented.

  2. Elemental imaging of rat epididymis by micro-PIXE analysis

    Science.gov (United States)

    Homma-Takeda, S.; Nishimura, Y.; Watanabe, Y.; Imaseki, H.; Yukawa, M.

    2003-09-01

    The epididymis, a male reproductive organ, which is a highly convoluted duct, plays an important role in transportation of spermatozoa, their maturation, and their storage. Although major elements, such as P, S and K, as well as trace elements, such as Mn, Cu, Zn, Se, are known to be essential for spermatogenesis, detailed distributions of the elements in the epididymis are only poorly understood. In the present study, Mn, Cu, Zn and Se levels in the epididymis were examined in male Wistar rats by inductively coupled argon plasma-mass spectrometry (ICP-MS) analysis and in situ multi-elemental distributions of epididymal sections were determined by micro-PIXE (particle induced X-ray emission) analysis. The Zn, Cu and Se concentrations in the epididymis of the young adult rats were around 30 μg/g wet weight, 2 μg/g wet weight and 1 μg/g wet weight, respectively, and their Mn were less than 0.5 μg/g wet weight. PIXE imaging of P and K exhibited that P and K were higher in the epididymal epithelium. In contrast, more S was detected in the lumen, which is composed of spermatozoa and a fluid. Elemental imagings of the trace elements were unclear compared with the major elements, but information about zinc localization in the epididymis was obtained.

  3. Elemental imaging of rat epididymis by micro-PIXE analysis

    Energy Technology Data Exchange (ETDEWEB)

    Homma-Takeda, S.; Nishimura, Y. E-mail: y_nishim@nirs.go.jp; Watanabe, Y.; Imaseki, H.; Yukawa, M

    2003-09-01

    The epididymis, a male reproductive organ, which is a highly convoluted duct, plays an important role in transportation of spermatozoa, their maturation, and their storage. Although major elements, such as P, S and K, as well as trace elements, such as Mn, Cu, Zn, Se, are known to be essential for spermatogenesis, detailed distributions of the elements in the epididymis are only poorly understood. In the present study, Mn, Cu, Zn and Se levels in the epididymis were examined in male Wistar rats by inductively coupled argon plasma-mass spectrometry (ICP-MS) analysis and in situ multi-elemental distributions of epididymal sections were determined by micro-PIXE (particle induced X-ray emission) analysis. The Zn, Cu and Se concentrations in the epididymis of the young adult rats were around 30 {mu}g/g wet weight, 2 {mu}g/g wet weight and 1 {mu}g/g wet weight, respectively, and their Mn were less than 0.5 {mu}g/g wet weight. PIXE imaging of P and K exhibited that P and K were higher in the epididymal epithelium. In contrast, more S was detected in the lumen, which is composed of spermatozoa and a fluid. Elemental imagings of the trace elements were unclear compared with the major elements, but information about zinc localization in the epididymis was obtained.

  4. Gait analysis in rats with peripheral nerve injury.

    Science.gov (United States)

    Yu, P; Matloub, H S; Sanger, J R; Narini, P

    2001-02-01

    Rats are commonly used to study peripheral nerve repair and grafting. The traditional footprint method to assess functional recovery is messy, indirect, and not useful when contractures develop in the animal model. The aim of the present study was to establish an accurate, reproducible, but simple, method to assess dynamic limb function. The basic quantitative aspects of a normal gait were characterized from 59 recorded walks in 23 rats. The video was digitized and analyzed frame by frame on a personal computer. Seven parameters of the gait were assessed: (1) walking speed; (2) stance phase, swing phase and right to left stance/swing ratio; (3) step length and step length ratio; (4) ankle angles at terminal stance and midswing; (5) tail height; (6) midline deviation; and (7) tail deviation. These gait parameters were then applied to groups of animals with sciatic (group S), tibial (group T), and peroneal (group P) nerve injuries. A discriminant analysis was performed to analyze each parameter and to compute a functional score. We found that the video gait analysis was superior to the footprint method and believe it will be very useful in future studies on peripheral nerve injury.

  5. Development and application of a UPLC-MS/MS method for simultaneous determination of fenofibric acid and berberine in rat plasma: application to the drug-drug pharmacokinetic interaction study of fenofibrate combined with berberine after oral administration in rats.

    Science.gov (United States)

    Li, Guofei; Yang, Fan; Liu, Mei; Su, Xianying; Zhao, Mingming; Zhao, Limei

    2016-07-01

    With the purpose of carrying out pharmacokinetic interaction studies ofnberberine (BBR) and fenofibrate (FBT), an UPLC-MS/MS method has been developed and validated. The analytes, BBR and fenofibric acid (FBA, metabolite of FBT) and the internal standard, tetrahydropalmatine, were extracted with dichloromethane-diethyl ether (3:2, v/v) and separated on an Agilent Eclipse XDB C18 column using a mobile phase composed of acetonitrile and water. With positive ion electrospray ionization, the analytes were monitored on a triple quadrupole mass spectrometer in multiple reaction monitoring mode. Linear calibration curves were obtained over the concentration ranges of 0.1-100.0 ng/mL for BBR and 10.0-50,000.0 ng/mL for FBA. For BBR and FBA, the intra- and inter-day precisions were 85.6%, and those of FBA at three concentrations of 20.0, 2500.0, 40,000.0 ng/mL were >87.9%. Consequently, the proposed method was applied to the pharmacokinetic interaction study of FBT combined with BBR after oral administration in rats and was proved to be sensitive, specific and reliable to analyze BBR and FBA in biological samples simultaneously. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Fast, multi-dimensional and simultaneous kymograph-like particle dynamics (SkyPad analysis.

    Directory of Open Access Journals (Sweden)

    Bruno Cadot

    Full Text Available BACKGROUND: Kymograph analysis is a method widely used by researchers to analyze particle dynamics in one dimensional (1D trajectories. RESULTS: Here we provide a Visual Basic-coded algorithm to use as a Microsoft Excel add-in that automatically analyzes particles in 2D trajectories with all the advantages of kymograph analysis. CONCLUSIONS: This add-in, which we named SkyPad, leads to significant time saving and higher accuracy of particle analysis. Finally, SkyPad can also be used for 3D trajectories analysis.

  7. Chromatographic fingerprint analysis and simultaneous determination of eight lignans in Justicia procumbens and its compound preparation by HPLC-DAD.

    Science.gov (United States)

    Wang, Linan; Pan, Jianyu; Yang, Meihua; Wu, Jun; Yang, Junshan

    2011-03-01

    HPLC fingerprints were developed for the quality evaluation of Justicia procumbens and its compound preparation, Jian-er syrup, together with the simultaneous quantification of eight arylnaphthalide lignans (6'-hydroxy justicidin B, 6'-hydroxy justicidin A, 6'-hydroxy justicidin C, justicidin B, chinensinaphthol methyl ether, justicidin C, taiwanin C, and neojusticin A). Samples were separated with a Shiseido Capcell Pak C(18) reversed-phase column (250×4.6 mm id, 5 μm) using acetonitrile and water as the mobile phase. The column temperature was maintained at 35°C and the wavelength of detector was set at 256 nm. For fingerprint analysis, 17 peaks were selected as the characteristic peaks for the evaluation of the similarities among different J. procumbens samples collected in different places. The structures of lignans were confirmed by diagnostic fragments in the positive ESI-MS(n) . The new method was successfully applied for the chromatographic fingerprint analysis and simultaneous determination of eight lignans in its compound preparation, Jian-er syrup. All the results indicated that HPLC fingerprint assay in combination with multi-marker determination afforded a useful method for the quality control of J. procumbens and its compound preparation, Jian-er syrup.

  8. Efficiency and Safety of Intravenous Tranexamic Acid in Simultaneous Bilateral Total Knee Arthroplasty: A Systematic Review and Meta-analysis.

    Science.gov (United States)

    Jiang, Xuan; Ma, Xin-Long; Ma, Jian-Xiong

    2016-08-01

    The objective of this systematic review and meta-analysis was to evaluate the efficacy and safety of i.v. tranexamic acid (TXA) in simultaneous bilateral total knee arthroplasty (TKA). Potentially relevant published reports were identified from the following electronic databases: Medline, PubMed, Embase, ScienceDirect and Cochrane Library. RevMan v5.3was used to pool data. Two randomized controlled trials and four case-control studies met the inclusion criteria. The current meta-analysis identified significant differences between TXA group and control groups in terms of postoperative hemoglobin concentration (P < 0.01), drainage volume (P < 0.01), transfusion rate (P < 0.01) and units transfused (P = 0.006). There were no significant differences in length of stay (P = 0.66), operation time (P = 0.81) or and incidence of adverse effects such as infection (P = 0.42), deep venous thrombosis (DVT) (P = 0.88) and pulmonary embolism (PE) (P = 0.11). Our results show that i.v. administration of TXA in simultaneous bilateral TKA reduces postoperative drops in hemoglobin concentration, drainage volume, and transfusion requirements and does not prolong length of stay or operation time. Moreover, no adverse effects, such as infection, DVT or PE, were associated with TXA.

  9. Simultaneous analysis of glycolipids and phospholids molecular species in avocado (Persea americana Mill) fruit.

    Science.gov (United States)

    Pacetti, Deborah; Boselli, Emanuele; Lucci, Paolo; Frega, Natale G

    2007-05-25

    The molecular species of phospholipids (PLs) and glycolipids (GLs) were simultaneously characterized in the pulp and almond of the avocado fruit (Persea americana Mill) of four varieties by means of high performance liquid chromatography-electrospray ionisation ion-trap tandem mass spectrometry. In the pulp, the predominant species of monoglycosyldiglycerides (MGD) were m/z 796.6 (oleic/linolenic and linoleic/linoleic acids) and m/z 800.4 (stearic/linoleic and oleic/oleic acids). One of the main diglycosyldiglycerides (DGD) both in the pulp and almond was m/z 958.5 (oleic/linolenic); however, the pulp was also rich of m/z 962.4 (oleic/oleic), whereas in the almond, m/z 934.5 (palmitic/linoleic and palmitoleic/oleic) and m/z 960.5 (oleic/linoleic and stearic/linolenic) were more abundant. In the almond, the main PL classes (phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI)) contained always palmitic/linoleic acids. Alpha-linolenic acid was contained as MGD (linolenic/linolenic) and DGD (linolenic/linolenic), more present in the pulp than in the almond. The major molecular species of glycocerebrosides (GCer) in the pulp and almond carried hydroxy-palmitic acid (C(16h:0))/4,8-sphyngadienine (d(18:2)).

  10. Measuring the pulse of GRB 090618: A Simultaneous Spectral and Timing Analysis of the Prompt Emission

    CERN Document Server

    RupalBasak,

    2011-01-01

    We develop a new method for simultaneous timing and spectral studies of Gamma Ray Burst (GRB) prompt emission and apply it to make a pulse-wise description of the prompt emission of GRB 090618, the brightest GRB detected in the Fermi era. We exploit the large area (and sensitivity) of Swift/BAT and the wide band width of Fermi/GBM to derive the parameters for a complete spectral and timing description of the individual pulses of this GRB, based on the various empirical relations suggested in the literature. We demonstrate that this empirical model correctly describes the other observed properties of the burst like the variation of the lag with energy and the pulse width with energy. The measurements also show an indication of an increase in pulse width as a function of energy at low energies for some of the pulses, which is naturally explained as an off-shoot of some particular combination of the model parameters. We argue that these model parameters, particularly the peak energy at the beginning of the pulse...

  11. A Simple and Sensitive HPLC Method for Simultaneous Analysis of Nabumetone and Paracetamol in Pharmaceutical Formulations

    Directory of Open Access Journals (Sweden)

    Prafulla Kumar Sahu

    2011-01-01

    Full Text Available This paper describes a high-performance liquid chromatographic method for simultaneous estimation of nabumetone and paracetamol in binary mixture. The method was based on RP-HPLC separation and quantitation of the two drugs on hypersil C-18 column (250 mm × 4.6 mm using a mobile phase consisting of acetonitrile and 0.05% aqueous acetic acid (70:30v/v at flow rate of 1 mL min-1. Quantitation was achieved with PDA detector at 238 nm based on peak area with linear calibration curves at concentration ranges 5-25 µg mL-1 for both the drugs. Naproxen sodium was used as internal standard. The method has been successively applied to pharmaceutical formulation. No chromatographic interference from the tablet excipients was found. The method was validated in terms of precision, robustness, recovery and limits of detection and quantitation. The intra and inter-day precision and accuracy values were in the acceptance range as per ICH guidelines.

  12. Simultaneous analysis of skin penetration of surfactant and active drug from fluorosurfactant-based microemulsions.

    Science.gov (United States)

    Mahrhauser, Denise; Hoppel, Magdalena; Schöll, Judith; Binder, Lisa; Kählig, Hanspeter; Valenta, Claudia

    2014-09-01

    The purpose of this study was to investigate the penetrated amount of the incorporated model drug diclofenac-sodium and of a fluorosurfactant as specific vehicle constituent of topically applied microemulsions at the same time. To this end, the penetration depth of each compound was elucidated through tape stripping studies by the simultaneous quantification of diclofenac-sodium and the fluorosurfactant from the same sample. A new approach was made by using the very sensitive and specific (19)F NMR (nuclear magnetic resonance) for quantification of the fluorinated vehicle component. The tape stripping experiments with the microemulsions showed an almost similar penetration velocity of diclofenac-sodium and fluorosurfactant, suggesting that the surfactant within the microemulsion-structure intensified the stratum corneum uptake of the incorporated active constituent. Moreover, ATR-FTIR studies on porcine ear skin revealed significant shifts of the CH₂ stretching absorbances, which are associated with an enhanced disorder of the SC lipids resulting in a decreased skin barrier function, after application of the microemulsions. However, the application of pure fluorosurfactant did not cause any shifts in the CH₂ stretching absorbances. It can be thereby concluded that the prepared microemulsions exerted specific effects on skin integrity resulting in a "push" of diclofenac-sodium penetration.

  13. Simultaneous Determination of Preservatives in Dairy Products by HPLC and Chemometric Analysis

    Directory of Open Access Journals (Sweden)

    Fatemeh Zamani Mazdeh

    2017-01-01

    Full Text Available Cheese and yogurt are two kinds of nutritious dairy products that are used worldwide. The major preservatives in dairy products are sodium benzoate, potassium sorbate, and natamycin. The maximum permitted levels for these additives in cheese and yogurt are established according to Iranian national standards. In this study, we developed a method to detect these preservatives in dairy products by reversed phase chromatography with UV detection in 220 nm, simultaneously. This method was performed on C18 column with ammonium acetate buffer (pH=5 and acetonitrile (73 : 27 v/v as mobile phase. The method was carried out on 195 samples in 5 kinds of commercial cheeses and yogurts. The results demonstrated insufficient separation where limit of detection (LOD and limit of quantitation (LOQ ranged from 0.326 to 0.520 mg/kg and 0.989 to 1.575 mg/kg in benzoate and sorbate, respectively. The correlation coefficient of each calibration curve was mostly higher than 0.997. All samples contained sodium benzoate in various ranges. Natamycin and sorbate were detected in a remarkable amount of samples, while, according to Iranian national standard, only sorbate is permitted to be added in processed cheeses as a preservative. In order to control the quality of dairy products, determination of preservatives is necessary.

  14. Simultaneous Determination of Preservatives in Dairy Products by HPLC and Chemometric Analysis.

    Science.gov (United States)

    Zamani Mazdeh, Fatemeh; Sasanfar, Sima; Chalipour, Anita; Pirhadi, Elham; Yahyapour, Ghazal; Mohammadi, Armin; Rostami, Akram; Amini, Mohsen; Hajimahmoodi, Mannan

    2017-01-01

    Cheese and yogurt are two kinds of nutritious dairy products that are used worldwide. The major preservatives in dairy products are sodium benzoate, potassium sorbate, and natamycin. The maximum permitted levels for these additives in cheese and yogurt are established according to Iranian national standards. In this study, we developed a method to detect these preservatives in dairy products by reversed phase chromatography with UV detection in 220 nm, simultaneously. This method was performed on C18 column with ammonium acetate buffer (pH = 5) and acetonitrile (73 : 27 v/v) as mobile phase. The method was carried out on 195 samples in 5 kinds of commercial cheeses and yogurts. The results demonstrated insufficient separation where limit of detection (LOD) and limit of quantitation (LOQ) ranged from 0.326 to 0.520 mg/kg and 0.989 to 1.575 mg/kg in benzoate and sorbate, respectively. The correlation coefficient of each calibration curve was mostly higher than 0.997. All samples contained sodium benzoate in various ranges. Natamycin and sorbate were detected in a remarkable amount of samples, while, according to Iranian national standard, only sorbate is permitted to be added in processed cheeses as a preservative. In order to control the quality of dairy products, determination of preservatives is necessary.

  15. Simultaneous pancreas-kidney transplantation in a single center:10-year retrospective analysis

    Institute of Scientific and Technical Information of China (English)

    ZHENG Jian-ming; SONG Wen-li; TU Jin-peng; FENG Gang; MO Chun-bai; SHEN Zhong-yang

    2011-01-01

    Background Simultaneous pancreas-kidney transplantation (SPKT) is the best treatment option for diabetic patients with advanced chronic renal failure. The current study aimed to analyze the surgical indications, treatments and prognosis of SPKT.Methods We retrospectively analyzed 40 cases of SPKT performed between December 1999 and January 2010 in our center, including the survival rate, complications and the reasons of reoperation.Results Of all the 40 SPKT cases, the one-year survival rates of the recipients, kidney and pancreas transplant graft were 97.6%, 97.6% and 92.7%, while 97.6%, 91.1%, 92.7% at 3 years and 83.6%, 78.0%, 79.4% at 5 years, respectively.After SPKT, 10 patients need reoperation because of surgical complications (14 operations). The reoperation rate was 25%, including 2 patients (4 operations) with hematuria, 4 patients with abdominal hemorrhage, 2 patients (3 operations)with abdominal infection, 1 patient with pancreatic venous thrombosis, 1 patient with anastomotic leakage, and 1 patient with fistula.Conclusion Although SPKT provides a successful and effective treatment for diabetics with end-stage renal disease,how to reduce the complications of this treatment still need further effort.

  16. Simultaneous Analysis and Quantification of Markers of Manjisthadi Churna Using High Performance Thin Layer Chromatography

    Science.gov (United States)

    Patel, V. R.; Patel, R. K.

    2013-01-01

    Manjisthadi churna has been traditionally used in the Ayurvedic system of medicine and by traditional medical practices of India to treat hyperlipidemia. A rapid, simple and accurate method with high performance thin layer chromatography has been developed to standardised Manjisthadi churna using rubiadin, sennoside and ellagic acid as markers. Methanol extract of Manjisthadi churna were used for high performance thin layer chromatography on silica gel plates. The Rf of rubiadin, sennoside-A and ellagic acid were found to 0.48, 0.23 and 0.72, respectively with densitometric scanning at 280 nm and the calibration plot were linear in the range of 100-600 ng of markers. The correlation coefficients were higher than 0.99 were indicative of good linear dependence of peaks area on concentration. The rubiadin, sennoside-A and ellagic acid contents in Manjisthadi churna were found to be 0.014, 0.038 and 0.534% w/w, respectively. This method permits reliable quantification of rubiadin, sennoside-A and ellagic acid with good resolution and separation of the same from other constitutes of the extract of Manjisthadi churna. Recovery value from 95.66-102.33% showed the reliability and reproducibility of the method. The proposed high performance thin layer chromatography method for simultaneous quantification of markers in Manjisthadi churna can be used for routine quality testing. PMID:23901170

  17. Simultaneous ultrastructural analysis of fluorochrome-photoconverted diaminobenzidine and gold immunolabeling in cultured cells

    Directory of Open Access Journals (Sweden)

    M. Malatesta

    2013-09-01

    Full Text Available Diaminobenzidine photoconversion is a technique by which a fluorescent dye is transformed into a stably insoluble, brown, electrondense signal, thus enabling examination at both bright field light microscopy and transmission electron microscopy. In this work, a procedure is proposed for combining photoconversion and immunoelectron microscopy: in vitro cell cultures have been first submitted to photoconversion to analyse the intracellular fate of either fluorescent nanoparticles or photosensitizing molecules, then processed for transmission electron microscopy; different fixative solutions and embedding media have been used, and the ultrathin sections were finally submitted to post-embedding immunogold cytochemistry. Under all conditions the photoconversion reaction product and the target antigen were properly detected in the same section; Epon-embedded, osmicated samples required a pre-treatment with sodium metaperiodate to unmask the antigenic sites. This simple and reliable procedure exploits a single sample to simultaneously localise the photoconversion product and a variety of antigens allowing a specific identification of subcellular organelles at the ultrastructural level.

  18. Comparative Performance Analysis of Mobile IPv6 Protocols: Special Reference to Simultaneous Bindings

    Directory of Open Access Journals (Sweden)

    Shariq Haseeb

    2006-01-01

    Full Text Available Mobile IP is the Internet Engineering Task Force (IETF proposal to cater for All-Internet Protocol (All-IP mobility. It forms the backbone for next generation Wireless Internet Technology to provide uninterrupted network service while on the move. Our paper conducts a performance study of the various Mobile Internet Protocol version 6 (IPv6 protocols such as Simple Mobile IPv6, Hierarchical Mobile IPv6, Fast handover Mobile IPv6, their combination and Simultaneous Bindings Mobile IPv6. The paper benchmarks the protocol variations against the standard Mobile IPv6 protocol, by studying them under Quality of Service (QoS parameters. We propose an evaluation model containing 5 mobile nodes and then gradually (5 nodes per stage increasing the mobile nodes to 50. The proposed network model is then simulated in an open source simulator NS-2. This paper goes further to propose the most suitable variation of the protocol to use and the challenges faced in deployment.

  19. Simultaneous Explanation of the $R_K$ and $R_{D^{(*)}}$ Puzzles: a Model Analysis

    CERN Document Server

    Bhattacharya, Bhubanjyoti; Guévin, Jean-Pascal; London, David; Watanabe, Ryoutaro

    2016-01-01

    $R_K$ and $R_{D^{(*)}}$ are two $B$-decay measurements that presently exhibit discrepancies with the SM. Recently, using an effective field theory approach, it was demonstrated that a new-physics model can simultaneously explain both the $R_K$ and $R_{D^{(*)}}$ puzzles. There are two UV completions that can give rise to the effective Lagrangian: (i) $VB$: a vector boson that transforms as an $SU(2)_L$ triplet, as in the SM, (ii) $U_1$: an $SU(2)_L$-singlet vector leptoquark. In this paper, we show that, in fact, $VB$ and $U_1$ can be distinguished experimentally. $VB$ also contributes to $\\tau \\to 3\\mu$, while $U_1$ does not. $VB$ predicts $1.9 \\times 10^{-10} \\le {\\cal B}(\\tau^-\\to\\mu^-\\mu^+\\mu^-) \\le 2.1 \\times 10^{-8}$, which should be measurable at Belle II (and perhaps LHCb). $\\tau \\to 3\\mu$ is therefore a smoking-gun signal of $VB$. For $U_1$, there are several observables that may point to this model. Perhaps the most interesting is the lepton-flavor-violating decay $\\Upsilon(3S) \\to \\mu \\tau$, which h...

  20. Towards an integrated biosensor array for simultaneous and rapid multi-analysis of endocrine disrupting chemicals

    Energy Technology Data Exchange (ETDEWEB)

    Scognamiglio, Viviana, E-mail: viviana.scognamiglio@mlib.ic.cnr.it [IC-CNR Istituto di Cristallografia, AdR1 Dipartimento Agroalimentare - Via Salaria Km 29.3 00015, Rome (Italy); Pezzotti, Italo; Pezzotti, Gianni; Cano, Juan; Manfredonia, Ivano [Biosensor S.r.l. - Via degli Olmetti 44 00060 Formello, Rome (Italy); Buonasera, Katia [IC-CNR Istituto di Cristallografia, AdR1 Dipartimento Agroalimentare - Via Salaria Km 29.3 00015, Rome (Italy); Arduini, Fabiana; Moscone, Danila; Palleschi, Giuseppe [Universita di Roma Tor Vergata, Dipartimento di Scienze e Tecnologie Chimiche - Via della Ricerca Scientifica 00133, Rome (Italy); Giardi, Maria Teresa [IC-CNR Istituto di Cristallografia, AdR1 Dipartimento Agroalimentare - Via Salaria Km 29.3 00015, Rome (Italy)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer A multitask biosensor for the detection of endocrine disrupting chemicals is proposed. Black-Right-Pointing-Pointer The sensing system employ an array of biological recognition elements. Black-Right-Pointing-Pointer Amperometric and optical transduction methods are provided in an integrated biosensor together with flow control systems. Black-Right-Pointing-Pointer The biosensing device results in an integrated, automatic and portable system for environmental and agrifood application. - Abstract: In this paper we propose the construction and application of a portable multi-purpose biosensor array for the simultaneous detection of a wide range of endocrine disruptor chemicals (EDCs), based on the recognition operated by various enzymes and microorganisms. The developed biosensor combines both electrochemical and optical transduction systems, in order to increase the number of chemical species which can be monitored. Considering to the maximum residue level (MRL) of contaminants established by the European Commission, the biosensor system was able to detect most of the chemicals analysed with very high sensitivity. In particular, atrazine and diuron were detected with a limit of detection of 0.5 nM, with an RSD% less than 5%; paraoxon and chlorpyrifos were revealed with a detection of 5 {mu}M and 4.5 {mu}M, respectively, with an RSD% less than 6%; catechol and bisphenol A were identified with a limit of detection of 1 {mu}M and 35 {mu}M respectively, with an RSD% less than 5%.

  1. Improved simultaneous gas-chromatographic analysis for homovanillic acid and vanillylmandelic acid in urine.

    Science.gov (United States)

    Leiendecker-Foster, C; Freier, E F

    1981-12-01

    We describe an improved gas-chromatographic method for the simultaneous quantitation of the catecholamine metabolites, homovanillic acid (3-methoxy-4-hydroxyphenylacetic acid) and vanillylmandelic acid (3-methoxy-4-hydroxymandelic acid). Our improvements in the method of Muskiet et al. (Clin. Chem. 23: 863, 1977) include a shorter program time and a longer silylation interval. Recovery and precision data obtained by this improved technique are similar to those of Muskiet et al. Vanillylmandelic acid results (y) were compared with those by the method of Pisano et al. (Clin. Chim. Acta 7: 285, 1962). The relation is expressed by the equation y = 0.52 + 1.05x (Sy . x = 2.33 mg/24 h and r = 0.997). Results for homovanillic acid (y) were compared with those by the method of Knight and Haymond (Clin. Chem. 23: 2007, 1977); the equation was y = 0.84 + 0.90x (Sy . x = 2.04 and r = 0.97). Retention times are also reported for several phenolic acids and other related compounds found in urine.

  2. Joint Procrustes Analysis for Simultaneous Nonsingular Transformation of Component Score and Loading Matrices

    Science.gov (United States)

    Adachi, Kohei

    2009-01-01

    In component analysis solutions, post-multiplying a component score matrix by a nonsingular matrix can be compensated by applying its inverse to the corresponding loading matrix. To eliminate this indeterminacy on nonsingular transformation, we propose Joint Procrustes Analysis (JPA) in which component score and loading matrices are simultaneously…

  3. The MAM-E17 schizophrenia rat model: Comprehensive behavioral analysis of pre-pubertal, pubertal and adult rats.

    Science.gov (United States)

    Kállai, Veronika; Tóth, Attila; Gálosi, Rita; Péczely, László; Ollmann, Tamás; Petykó, Zoltán; László, Kristóf; Kállai, János; Szabó, Imre; Karádi, Zoltán; Lénárd, László

    2017-08-14

    The MAM-E17 model is one of the most accepted schizophrenia rat models, which follows the neurodevelopmental theory of the disease. While symptoms of MAM-E17 rats were studied extensively, their examinations were usually restricted to adulthood and in a few cases to prepuberty. It is well known, however, that schizophrenia symptoms often start at puberty or early adulthood. Therefore the purpose of this study was to investigate the behavioral characteristics of MAM-E17 rats in various tests throughout three different age-periods, namely in prepuberty, late puberty and adulthood. In open field test, MAM-E17 rats displayed increased locomotor activity, elevated sniffing frequency and, as tendency, enhanced rearing activity. The elevated activity turned up in late puberty and remained there in adulthood, too. There was also a deficient prepulse inhibition (PPI) of startle response in late puberty and adulthood, but not before puberty. In rotarod task, MAM-treated rats performed better than control rats. The enhanced performance on rotarod was only present in late puberty and adulthood. In elevated plus maze test MAM-treated rats displayed diminished anxiety mostly in prepuberty. Histological analysis revealed reduced volume and cell disarray in the dorsal hippocampus. This is the first comprehensive study about symptoms of MAM-E17 rats manifested in behavioral tests carried out in prepuberty, late puberty and adulthood. Results display the age-dependent appearance of schizophrenia symptoms in the same rats. The present findings provide basic information to accomplish the schizophrenia related animal research, as well as can also confer further data to develop preventive treatment for human patients. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Simultaneous hepatectomy and splenectomy versus hepatectomy alone for hepatocellular carcinoma complicated by hypersplenism: a meta-analysis

    Directory of Open Access Journals (Sweden)

    Li W

    2015-08-01

    Full Text Available Wei Li, Shi-Qiang Shen, Shan-Min Wu, Zu-Bing Chen, Chao Hu, Rui-Chen YanDepartment of General Surgery, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of ChinaBackground: This study systematically compared the efficacy and safety of simultaneous hepatectomy and splenectomy (HS with hepatectomy (H alone in patients with hepatocellular carcinoma (HCC and hypersplenism.Methods: The PubMed, Web of Science, Science Direct, EMBASE, and Cochrane Library databases were systematically searched by two independent researchers through to March 31, 2015 to identify relevant studies. All the extracted literature were managed by Bibliographic citation management software. Quality assessment of the included studies was performed using a modified Newcastle–Ottawa Scale judgment. The data were analyzed using RevMan5.2 software.Results: Eight studies including a total of 761 patients with HCC and hypersplenism (360 in the HS group, 401 in the H group were finally included in the analysis. Outcomes, including postoperative complications, perioperative mortality, operation time, 5-year survival rate, and need for blood transfusion did not differ significantly between the two groups. HS was associated with significantly more intraoperative bleeding (mean difference [MD] 57.15, 95% confidence interval [CI] 18.83–95.46, P=0.003, and CD4/CD8 ratio (MD 0.69, 95% CI 0.61–0.77, P<0.00001, CD4 subset, platelet count (MD 213.06, 95% CI 202.59–223.53, P<0.0001, white blood cell count (MD 4.85, 95% CI 4.58–5.13, P<0.0001, interferon-gamma levels (MD 18.52, 95% CI 13.93–23.11, P<0.00001, and interleukin-2 levels (MD 20.73, 95% CI 16.05–25.41, P<0.0001. In addition, lower CD8 subset (MD −7.85, 95% CI −9.07, −6.63, P<0.00001 and interleukin-10 levels (MD −18.56, 95% CI −22.61, −14.50, P<0.00001 were observed for HS.Conclusion: We identified that simultaneous HS do not increase postoperative complications, operation time, or perioperative

  5. Assessment of intramural blood flow and neurogenic control in intact and hypertrophic urinary bladder with harmonic analysis of bioimpedance in rats.

    Science.gov (United States)

    Kirpatovsky, V I; Mudraya, I S; Revenko, S V; Nesterov, A V; Gavrilov, I Yu; Khromov, R A; Bablumyan, A Yu

    2012-08-01

    High-resolution impedancometry and harmonic (Fourier) analysis of variable component of bioimpedance revealed rhythmic oscillations of urinary bladder bioimpedance at the Mayer wave, respiration, and heartbeat frequencies. The power values of the corresponding Mayer, respiratory, and cardiac peaks were calculated to assess circulation in the urinary bladder wall and its autonomic nervous control at various stages of infusion cystometry in intact rats and in the rats with preliminary formed infravesical obstruction (IVO). In intact rats, filling of the bladder with physiological saline diminished the power of the first (fundamental) cardiac peak attesting to a decrease of the blood flow in the bladder wall. Simultaneously, the power of low-frequency Mayer peak reflecting sympathetic activity increased, while the power of respiratory peak decreased supposedly reflecting abatement of the parasympathetic influences. Bladder voiding was accompanied by a decrease of Mayer peak and increase of the respiratory one. Prior to infusion cystometry, the intravesical pressure in IVO rats was elevated while the power of fundamental cardiac peak was below the control value. Filling the bladder in these rats was accompanied by further decrease of the cardiac peak reflecting still greater drop in blood supply. In control rats, voiding the bladder normalized the vesical circulation assessed by the cardiac peak, while in IVO rats this peak remained decreased. The reciprocal changes of Mayer and respiratory peaks observed during infusion cystometry in the norm were replaced by unidirectional decrease in the power of both peaks in IVO rats, which probably attest to disturbance of autonomic nervous control in the hypertrophic urinary bladder in these rats.

  6. Simultaneous Analysis of the 2nu2, nu1, and nu3 Bands of Hydrogen Telluride

    Science.gov (United States)

    Flaud; Betrencourt; Arcas; Burger; Polanz; Lafferty

    1997-04-01

    Spectra of a natural sample of hydrogen telluride as well as a spectrum of monoisotopic H2 130Te have been recorded by means of Fourier transform spectrometry with a resolution of 0.003 cm-1 in the spectral domain 7.5-4.3 μm where it is easy to observe the main absorbing bands nu1 and nu3. We have located and assigned for the first time the 2nu2 band which appears in the lower wavenumber range of the recorded spectral domain near 1700 cm-1. It proved necessary to treat simultaneously the three states (020), (100), and (001). nu1 and nu3 are indeed Coriolis-coupled vibration-rotation bands and it was observed that a few rotational levels of (001) could not be fitted to within their experimental accuracy without considering the second-order Coriolis interaction between the rotational levels of (020) and (001). In this way all the experimental levels were calculated to within the experimental uncertainty, and precise sets of vibrational energies and rotational and coupling constants were obtained for the seven most abundant H2Te isotopic species, namely H2 130Te, H2 128Te, H2 126Te, H2 125Te, H2 124Te, H2 123Te, and H2 122Te. For the most abundant isotopic species H2 130Te the bands centers arenu0 (2nu2) = 1715.9568, nu0 (nu1) = 2065.2709, nu0 (nu3) = 2072.1101 cm-1. Copyright 1997Academic Press

  7. Simultaneous liver and kidney transplantation:analysis of a single-center experience

    Institute of Scientific and Technical Information of China (English)

    MA Yi; WANG Guo-dong; HE Xiao-shun; LI Qiang; LI Jun-liang; ZHU Xiao-feng; WANG Chang-xi

    2010-01-01

    Background Simultaneous liver and kidney transplantation (SLKT) has been proven to be a favorable treatment for combined renal and hepatic end-stage disease.However, recipients receiving SLKT have a long medical history, poor general condition that is often accompanied by anemia, hypoalbuminemia, coagulopathy, water-electrolyte imbalance and acid-base disorders.This study aimed to explore the indications, surgical techniques, therapeutic experience,prevention and treatment of postoperative complications of SLKT.Methods The clinical data of 22 SLKTs cases performed at the First Affiliated Hospital of Sun Yat-sen University from January 2001 to December 2008 were retrospectively studied.Indications for SLKT, surgical techniques, perioperative fluid management, immunosuppressive regimen and experience in prevention and treatment of postoperative complications were analyzed.Results All operations were successfully performed.Postoperative complications occurred in 13 cases (59.1%), including pleural effusions (7), intra-abdominal bleeding (2), biliary complications (2), repeated upper gastrointestinal bleeding (1), and acute liver graft rejection (1).All complications were treated conservatively.In this study, there were five deaths during follow-up, in which three perioperative deaths occurred due to serious conditions.Mortality at 3 months was 13.6%.The one and three year patient survival rate was 81.3% and 73.9% respectively.Conclusions SLKT is an effective therapy for end-stage liver disease with chronic renal failure or severe damage to renal function.It is a complex surgical procedure, causing a large disturbance of circulation and fluid balance, and more postoperative complications.The SLKT surgical techniques selected are based on the experience of surgeons, the anatomy of the recipient and primary diseases.It is essential to use the correct perioperative fluid management, reasonable immunosuppressive regimen, and prevention and treatment of postoperative

  8. Simultaneous measurement of denitrification and nitrogen fixation using isotope pairing with membrane inlet mass spectrometry analysis.

    Science.gov (United States)

    An, S; Gardner, W S; Kana, T

    2001-03-01

    A method for estimating denitrification and nitrogen fixation simultaneously in coastal sediments was developed. An isotope-pairing technique was applied to dissolved gas measurements with a membrane inlet mass spectrometer (MIMS). The relative fluxes of three N(2) gas species ((28)N(2), (29)N(2), and (30)N(2)) were monitored during incubation experiments after the addition of (15)NO(3)(-). Formulas were developed to estimate the production (denitrification) and consumption (N(2) fixation) of N(2) gas from the fluxes of the different isotopic forms of N(2). Proportions of the three isotopic forms produced from (15)NO(3)(-) and (14)NO(3)(-) agreed with expectations in a sediment slurry incubation experiment designed to optimize conditions for denitrification. Nitrogen fixation rates from an algal mat measured with intact sediment cores ranged from 32 to 390 microg-atoms of N m(-2) h(-1). They were enhanced by light and organic matter enrichment. In this environment of high nitrogen fixation, low N(2) production rates due to denitrification could be separated from high N(2) consumption rates due to nitrogen fixation. Denitrification and nitrogen fixation rates were estimated in April 2000 on sediments from a Texas sea grass bed (Laguna Madre). Denitrification rates (average, 20 microg-atoms of N m(-2) h(-1)) were lower than nitrogen fixation rates (average, 60 microg-atoms of N m(-2) h(-1)). The developed method benefits from simple and accurate dissolved-gas measurement by the MIMS system. By adding the N(2) isotope capability, it was possible to do isotope-pairing experiments with the MIMS system.

  9. STATISTICAL METHODOLOGY FOR THE SIMULTANEOUS ANALYSIS OF MULTIPLE TYPES OF OUTCOMES IN NONLINEAR THRESHOLD MODELS.

    Science.gov (United States)

    Multiple outcomes are often measured on each experimental unit in toxicology experiments. These multiple observations typically imply the existence of correlation between endpoints, and a statistical analysis that incorporates it may result in improved inference. When both disc...

  10. Simultaneous determination of salidroside and its aglycone metabolite p-tyrosol in rat plasma by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Guo, Na; Hu, Zhiwei; Fan, Xiaoxu; Zheng, Jian; Zhang, Dehui; Xu, Tao; Yu, Tao; Wang, Yang; Li, Haiying

    2012-04-23

    Salidroside and its aglycone p-tyrosol are two major phenols in the genus Rhodiola and have been confirmed to possess various pharmacological properties. In our present study, p-tyrosol was identified as the deglycosylation metabolite of salidroside after intravenous (i.v.) administration to rats at a dose of 50 mg/kg, but was not detectable after intragastric gavage (i.g.) administration through HPLC-photodiode array detection (PDA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Next, an accurate and precise LC-MS/MS method was developed to quantitatively determine salidroside and p-tyrosol in rat plasma samples. Samples were analyzed by LC-MS/MS on a reverse-phase xTerra MS C18 column which was equilibrated and eluted with an isocratic mixture of acetonitrile-water (1:9, v/v) at a flow rate of 0.3 mL/min. The analytes were monitored by multiple reaction monitoring (MRM) under the negative electrospray ionization mode. The precursor/product transitions (m/z) were 299.0 → 118.8 for salidroside, 137.0 → 118.9 for p-tyrosol and 150.1 → 106.9 for the internal standard (IS), paracetamol, respectively. The calibration curve was linear over the concentration ranges of 50-2,000 ng/mL for salidroside and 20-200 ng/mL for p-tyrosol. The inter- and intra-day accuracy and precision were within ± 15%. The method has been successfully applied to the pharmacokinetic study and the oral bioavailability was calculated.

  11. Simultaneous Determination of Salidroside and Its Aglycone Metabolite p-Tyrosol in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Na Guo

    2012-04-01

    Full Text Available Salidroside and its aglycone p-tyrosol are two major phenols in the genus Rhodiola and have been confirmed to possess various pharmacological properties. In our present study, p-tyrosol was identified as the deglycosylation metabolite of salidroside after intravenous (i.v. administration to rats at a dose of 50 mg/kg, but was not detectable after intragastric gavage (i.g. administration through HPLC-photodiode array detection (PDA and liquid chromatography-tandem mass spectrometry (LC-MS/MS analysis. Next, an accurate and precise LC-MS/MS method was developed to quantitatively determine salidroside and p-tyrosol in rat plasma samples. Samples were analyzed by LC-MS/MS on a reverse-phase xTerra MS C18 column which was equilibrated and eluted with an isocratic mixture of acetonitrile-water (1:9, v/v at a flow rate of 0.3 mL/min. The analytes were monitored by multiple reaction monitoring (MRM under the negative electrospray ionization mode. The precursor/product transitions (m/z were 299.0→118.8 for salidroside, 137.0→118.9 for p-tyrosol and 150.1→106.9 for the internal standard (IS, paracetamol, respectively. The calibration curve was linear over the concentration ranges of 50–2,000 ng/mL for salidroside and 20–200 ng/mL for p-tyrosol. The inter- and intra-day accuracy and precision were within ±15%. The method has been successfully applied to the pharmacokinetic study and the oral bioavailability was calculated.

  12. New method for the simultaneous determination of pyrite content and proximate analysis in coal and lignite

    Energy Technology Data Exchange (ETDEWEB)

    Aylmer, D.M.

    1983-01-01

    The new method combines thermogravimetry and thermomagnetometry and utilizes inert, oxidizing, and reducing gases. Results by the new technique are compared to the ASTM method, one set obtained by the author on a Fisher Coal Analyzer and one set by the Coal Research Laboratory of the Pennsylvania State University. Comparison of thermo-magneto-gravimetric-analysis with the ASTM method indicates good agreement and comparable accuracy. These studies show that the thermo-magneto-gravimetric-analysis are: 1) ease of determination of both proximate analysis and pyrite, which permits use of unskilled technicians; 2) widespread availability of the apparatus; 3) cost effectiveness due to use of unskilled operators; 4) automation, presently available for proximate analysis on some commercial instruments and is easily accomplished for pyrite analysis, as well; 5) advantage over pyrite analysis by the ASTM method in two situations: first, when pyrite is totally surrounded by acid-insoluble organic-material and secondly where significant amounts of pyrite have been oxidized to FeSO/sub 4/; and 6) a permanent record of the measurements, which are continuous, is made in contrast to the ASTM method which records only initial and final conditions.

  13. Simultaneous transcriptome analysis of Sorghum and Bipolaris sorghicola by using RNA-seq in combination with de novo transcriptome assembly.

    Directory of Open Access Journals (Sweden)

    Takayuki Yazawa

    Full Text Available The recent development of RNA sequencing (RNA-seq technology has enabled us to analyze the transcriptomes of plants and their pathogens simultaneously. However, RNA-seq often relies on aligning reads to a reference genome and is thus unsuitable for analyzing most plant pathogens, as their genomes have not been fully sequenced. Here, we analyzed the transcriptomes of Sorghum bicolor (L. Moench and its pathogen Bipolaris sorghicola simultaneously by using RNA-seq in combination with de novo transcriptome assembly. We sequenced the mixed transcriptome of the disease-resistant sorghum cultivar SIL-05 and B. sorghicola in infected leaves in the early stages of infection (12 and 24 h post-inoculation by using Illumina mRNA-Seq technology. Sorghum gene expression was quantified by aligning reads to the sorghum reference genome. For B. sorghicola, reads that could not be aligned to the sorghum reference genome were subjected to de novo transcriptome assembly. We identified genes of B. sorghicola for growth of this fungus in sorghum, as well as genes in sorghum for the defense response. The genes of B. sorghicola included those encoding Woronin body major protein, LysM domain-containing intracellular hyphae protein, transcriptional factors CpcA and HacA, and plant cell-wall degrading enzymes. The sorghum genes included those encoding two receptors of the simple eLRR domain protein family, transcription factors that are putative orthologs of OsWRKY45 and OsWRKY28 in rice, and a class III peroxidase that is a homolog involved in disease resistance in the Poaceae. These defense-related genes were particularly strongly induced among paralogs annotated in the sorghum genome. Thus, in the absence of genome sequences for the pathogen, simultaneous transcriptome analysis of plant and pathogen by using de novo assembly was useful for identifying putative key genes in the plant-pathogen interaction.

  14. [Electrophysiological analysis of functional state of spinal cord motoneurons in rats with parathyroprivous tetany].

    Science.gov (United States)

    Khudaverdian, D N; Avetisian, K A; Chavushian, V A; Sarkisian, Dzh C

    2014-05-01

    In normal rats and of those with parathyroprivous (hypocalcemic) tetany the comparative analysis of background activity (BA), tetanic and posttetanic increase and decrease of frequency of spinal cord (SC) motoneurons (MNs) responses under high-frequency (50, 100Hz) stimulation (HFS) of flexor (G) and extensor (P) hind-limb nerves have been conducted. The on-line selection and program analysis of the spikes was produced. On the 3-7 and 21-22 days of development of acute and chronic tetany, respectively, the significant tetanic and posttetanic changes of MNs activity without meaningful changes in BA was registered. Along with the abrupt increase of excitatory manifestation of activity to HFS in a period of development of acute tetany was observed their relative weakening in animals with chronic tetany. Simultaneously the weakening or total disappearance of depressor reaction, especially expressed in the period of development of acute tetany was noted. It was concluded on the causal dependence of the parathyroprivous convulsions due to disturbances of correlation of inhibitory-excitatory processes in SC MNs.

  15. Analysis of femur head microstructure in ovariectomized rats

    Science.gov (United States)

    Andrade, C. B. V.; Nogueira, L. P.; Salata, C.; da Silva, C. M.; Ferreira-Machado, S. C.; de Almeida, C. E.; Almeida, A. P.; Colaço, M. V.; Alessio, R. C. P. V.; Braz, D.; Tromba, G.; Barroso, R. C.

    2013-07-01

    It is well accepted that rat ovariectomy (OVX) is a model of estrogen deficiency. OVX played a very important role in the initiating and developing of osteoporosis and it has been shown to be a major risk factor for the development of postmenopausal osteoporosis in women. In this work we used synchrotron radiation computed microtomography to investigate the skeletal effects in femoral head of female Wistar rats after bilateral ovariectomy surgery. The CT system was set up at the SYRMEP beamline in the synchrotron radiation facility ELETTRA (Trieste, Italy). Micro-CT images provided 3D information on precise trabecular microstructure by the reconstruction of multiple 2D images with almost 2 μm resolution. Our aim was to use histomorphometric analysis to reveal the effect of OVX on the three-dimensional (3D) trabecular bone microarchitecture. Evaluated morphometric parameters were trabecular bone volume-tissue volume ratio (BV/TV), trabecular number (Tb.N), trabecular separation (Tb.Sp) and trabecular thickness (Tb.Th). OVX group presented noticeable reduction in the Tb.N and Tb.Th when compared with control group (P 0.05). Our data may help to gain more insight into the potential mechanism of osteoporotic femoral head fractures.

  16. Simultaneous measurements of capillary filtration and diffusion capacities during graded infusions of noradrenaline (NA) and 5-hydroxytryptamine (5-HT) into the rat hindquarter vascular bed.

    Science.gov (United States)

    Rippe, B; Folkow, B

    1980-07-01

    The relationships between capillary diffusion capacity (PS) for Cr-EDTA respective capillary filtration capacity (CFC) and vascular resistance during graded intraarterial infusions of NA and 5-HT into the artificially constant flow perfused rat hindquarter vascular bed were investigated. During maximal vasodilatation PS for Cr-EDTA was some 5.5--5.7 ml/min x 100 g, CFC some 0.04 ml/min x mmHg x 100 g, while vascular resistance was 2.8 mmHg x ml-1 x min x 100 g (PRU100) and isogravimetric capillary pressure 12.8 mmHg on an average. Setting out from maximal vasodilatation, increasing doses of NA and 5-HT produced graded reductions in capillary surface area as reflected by progressive decreases in both PS for Cr-EDTA and CFC. These changes occurred simultaneously with progressive increases in both pre- and postcapillary resistances, causing elevations in both arterial and capillary hydrostatic pressures and hance in capillary fluid filtration at constant flow. Capillary hydrostatic pressure increased maximally to 45 mmHg (calculated for NA) and vascular resistance to some 21 mmHg x ml-1 x min x 100 g on an average. PS for Cr-EDTA decreased maximally to some 0.7--1 ml/min x 100 g for both NA and 5-HT and furthermore, the relationships between PS for Cr-EDTA and PRU100 for NA respective 5-HT were almost identical. This was taken to indicate that capillary surface area for nutritional exchange is affected similarly by both drugs. However, the CFU-PRU100 relationship was shifted towards some 30--50% higher CFC values for 5-HT than for NA at almost every level of vasoconstriction. This might suggest that 5-HT besides reducing capillary surface area also induced moderate increases in capillary permeability though increases in number and/or radius of large pores (gaps) (cf. Rippe, Kamiya & Folkow 1978). Even during NA-induced vasoconstriction, when virtually no changes in capillary permeability occurred, PS for Cr-EDTA was reduced to a relatively greater extent than CFC, the

  17. New approach to primary mass composition analysis with simultaneous use of ground and fluorescence detectors data

    CERN Document Server

    Yushkov, A; Aramo, C; Guarino, F; D'Urso, D; Valore, L

    2009-01-01

    We study the possibility to reconstruct primary mass composition with the use of combinations of basic shower characteristics, measured in hybrid experiments, such as depth of shower maximum from fluorescence side and signal in water Cherenkov tanks or in plastic scintillators from the ground side. To optimize discrimination performance of shower observables combinations we apply Fisher's discriminant analysis and give statistical estimates of separation of the obtained distributions on Fisher variables for proton and iron primaries. At the final stage we apply Multiparametric Topological Analysis to these distributions to extract composition from prepared mixtures with known fractions of showers from different primary particles. It is shown, that due to high sensitivity of water tanks to muons, combination of signal in them with $\\xmax$ looks especially promising for mass composition analysis, provided the energy is determined from longitudinal shower profile.

  18. The simultaneous separation and determination of six flavonoids and troxerutin in rat urine and chicken plasma by reversed-phase high-performance liquid chromatography with ultraviolet-visible detection.

    Science.gov (United States)

    Yang, Gong-Jun; Liu, Ping; Qu, Xi-Long; Xu, Min-Juan; Qu, Qi-Shu; Wang, Cheng-Yin; Hu, Xiao-Ya; Wang, Zhi-Yue

    2007-09-01

    The method of high-performance liquid chromatography (HPLC) with UV-vis detection was used and validated for the simultaneous determination of six flavonoids (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and troxerutin in rat urine and chicken plasma. Chromatographic separation was performed using a VP-ODS column (150 mm x 4.6 mm, 5.0 microm) maintained at 35.0 degrees C. The mobile phase was a mixture of water, methanol and acetic acid (57:43:1, v/v/v, pH 3.0) at the flow rate of 0.8 mL/min. Six flavonoids and troxerutin were analyzed simultaneously with good separation. On optimum conditions, calibration curves were found to be linear with the ranges of 0.10-70.00 microg/mL (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and 0.50-350.00 microg/mL (troxerutin). The detection limits were 0.010-0.050 microg/mL. The method was validated for accuracy and precision, and it was successfully applied to determine drug concentrations in rat urine and chicken plasma samples from rat and chicken that had been orally administered with six flavonoids and troxerutin.

  19. Analysis of amphetamines in urine with liquid-liquid extraction by capillary electrophoresis with simultaneous electrochemical and electrochemiluminescence detection.

    Science.gov (United States)

    Sun, Jinying; Xu, Xiaoyu; Wang, Chunyan; You, Tianyan

    2008-10-01

    Amphetamines including methamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylenedioxymethamphetamine were separated and detected by CE using simultaneous electrochemical (EC) and electrochemiluminescence (ECL) detection (CE-EC/ECL). Factors that influenced the separation and detection performance, such as the detection potential, the pH value and concentration of the running buffer, the separation voltage and the pH of the detection buffer, were investigated. LODs of 3.3x10(-8) mol/L (0.16 fmol), 1.6x10(-7) mol/L (0.78 fmol) and 3.3x10(-8) mol/L (0.16 fmol) were obtained for methamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylenedioxymethamphetamine, respectively. For practical application, a liquid-liquid extraction with ethyl acetate procedure was developed for urine sample pretreatment and extraction efficiencies higher than 90% were obtained. The established simultaneous CE-EC/ECL was successfully applied for urine sample analysis.

  20. Quad quantum cascade laser spectrometer with dual gas cells for the simultaneous analysis of mainstream and sidestream cigarette smoke.

    Science.gov (United States)

    Baren, Randall E; Parrish, Milton E; Shafer, Kenneth H; Harward, Charles N; Shi, Quan; Nelson, David D; McManus, J Barry; Zahniser, Mark S

    2004-12-01

    A compact, fast response, infrared spectrometer using four pulsed quantum cascade (QC) lasers has been applied to the analysis of gases in mainstream (MS) and sidestream (SS) cigarette smoke. QC lasers have many advantages over the traditional lead-salt tunable diode lasers, including near room temperature operation with thermoelectric cooling and single mode operation with improved long-term stability. The new instrument uses two 36 m, 0.3 l multiple pass absorption gas cells to obtain a time response of 0.1s for the MS smoke system and 0.4s for the SS smoke system. The concentrations of ammonia, ethylene, nitric oxide, and carbon dioxide for three different reference cigarettes were measured simultaneously in MS and SS smoke. A data rate of 20Hz provides sufficient resolution to determine the concentration profiles during each 2s puff in the MS smoke. Concentration profiles before, during and after the puffs also have been observed for these smoke constituents in SS smoke. Also, simultaneous measurements of CO(2) from a non-dispersive infrared (NDIR) analyzer are obtained for both MS and SS smoke. In addition, during this work, nitrous oxide was detected in both the MS and SS smoke for all reference cigarettes studied.

  1. Simultaneous quantification of amphetamine, opiates, ketamine and relative metabolites in urine for confirmatory analysis by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Lin, Huei-Ru; Choi, Ka-Ian; Lin, Tzu-Chieh; Hu, Anren

    2013-06-15

    The rise in amphetamine, ketamine and opiates abuse in Taiwan has created a need for a reliable confirmatory assay. A method that combines superficially porous liquid chromatography tandem mass spectrometry (LC-MS/MS) with solid-phase extraction (SPE) was developed for the simultaneous quantification of amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), ketamine, opiates, and their corresponding metabolites in urine. The total run time of the method was 6.7min including equilibration time. The method was validated in accordance with the European Commission (EC) Decision 2002/642/EC. The within- and between-day precision was below 13.6% and the accuracy ranged from -17.1% to +9.9% for all analytes. Ion suppression was observed but compensated by using deuterated internal standards. No carryover was detected and the analytes were stable at room temperature for 16h, and for 72h at 4°C, and three-thaw cycles. The method was further validated by comparison with a reference gas chromatography-mass spectrometry (GC-MS) method, using 52 authentic urine samples. The results indicated that for the target analytes studied, the LC-MS/MS analysis was as precise, accurate, and specific as the GC-MS method. In conclusion, the present LC-MS/MS method is robust and reliable, and suitable for use as a confirmation assay in the simultaneous urine drug testing and quantification of amphetamines, ketamines, and opiates.

  2. Optimisation and validation of analytical methods for the simultaneous extraction of antioxidants: application to the analysis of tomato sauces.

    Science.gov (United States)

    Motilva, Maria-José; Macià, Alba; Romero, Maria-Paz; Labrador, Agustín; Domínguez, Alba; Peiró, Lluís

    2014-11-15

    In the present study, simultaneous extraction of natural antioxidants (phenols and carotenoids) in complex matrices, such as tomato sauces, is presented. The tomato sauce antioxidant compounds studied were the phenolics hydroxytyrosol, from virgin olive oil, quercetin and its derivatives, from onions, and quercetin-rutinoside as well as the carotenoid, lycopene (cis and trans), from tomatoes. These antioxidant compounds were extracted simultaneously with n-hexane/acetone/ethanol (50/25/25, v/v/v). The phenolics were analysed by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS), and lycopene (cis- and trans-forms) was analysed using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD). After studying the parameters of these methods, they were applied to the analysis of virgin olive oil, fresh onion, tomato concentrate and tomato powder, and commercial five tomato sauces. Subsequently, the results obtained in our laboratory were compared with those from the Gallina Blanca Star Group laboratory.

  3. Stable isotope analysis as an early monitoring tool for community-scale effects of rat eradication

    Science.gov (United States)

    Nigro, Katherine M.; Hathaway, Stacie A.; Wegmann, Alex; Miller-ter Kuile, Ana; Fisher, Robert N.; Young, Hillary S.

    2017-01-01

    Invasive rats have colonized most of the islands of the world, resulting in strong negative impacts on native biodiversity and on ecosystem functions. As prolific omnivores, invasive rats can cause local extirpation of a wide range of native species, with cascading consequences that can reshape communities and ecosystems. Eradication of rats on islands is now becoming a widespread approach to restore ecosystems, and many native island species show strong numerical responses to rat eradication. However, the effect of rat eradication on other consumers can extend beyond direct numerical effects, to changes in behavior, dietary composition, and other ecological parameters. These behavioral and trophic effects may have strong cascading impacts on the ecology of restored ecosystems, but they have rarely been examined. In this study, we explore how rat eradication has affected the trophic ecology of native land crab communities. Using stable isotope analysis of rats and crabs, we demonstrate that the diet or trophic position of most crabs changed subsequent to rat eradication. Combined with the numerical recovery of two carnivorous land crab species (Geograpsus spp.), this led to a dramatic widening of the crab trophic niche following rat eradication. Given the established importance of land crabs in structuring island communities, particularly plants, this suggests an unappreciated mechanism by which rat eradication may alter island ecology. This study also demonstrates the potential for stable isotope analysis as a complementary monitoring tool to traditional techniques, with the potential to provide more nuanced assessments of the community- and ecosystem-wide effects of restoration.

  4. Simultaneous Monitoring of Glucose, Lactate and L-Glutamate in Rat Blood by a Flow-injection Enzyme Electrode Array System

    Institute of Scientific and Technical Information of China (English)

    万巧; 张芬芬; 刘梅川; 朱自强; 鲜跃仲; 金利通

    2005-01-01

    Rapid measurement of glucose, lactate and L-glutamate level in blood is important for studying the balance of energy in body. The flow-injection analysis (FIA) system with enzyme electrode array was based on neutral red-doped silica (NRDS) nanoparticles as electrocatalyst. These uniform NRDS nanoparticles (about 50±3 nm) were prepared by a water-in-oil (W/O) microemulsion method, and characterized by TEM technique. The doped inside neutral red maintained its high electron-activity, while the outside nano silica surface prevented neutral red from leaching out into the aqueous solutions and showed high biocompability. These nanoparticles were then mixed with the glucose oxidase (GOD), lactate oxidase (LOD) or L-glutamate oxidase (L-GLOD), and immobilized on a three carbon-disk electrode (CE) array, respectively. A thin Nation film was coated on the enzyme layer to prevent interference such as ascorbic acid and uric acid in the blood. The proposed flow-injection analysis with NRDS-enzyme electrode array method enables simultaneously monitoring various levels of glucose, lactate and L-glutamate in blood.

  5. Integration of XRF spectrometer for simultaneous and/or complementary use with PIXE at the external ion beam analysis setup

    Energy Technology Data Exchange (ETDEWEB)

    Fazinic, S.; Cosic, D.; Jaksic, M. [Laboratory for lon Beam Interactions, Division of Experimental Physics, Rudjer Boskovic Institute, Zagreb (Croatia); Migliori, A.; Karydas, A.G. [Nuclear Spectrometry and Applications Laboratory, International Atomic Energy Agency (IAEA), Vienna (Austria); Desnica, V. [Laboratory for Science and Technology in Art, Academy of Fine Arts in Zagreb, Zagreb (Croatia); Mudronja, D. [Natural Science Laboratory, Croatian Conservation Institute, Zagreb (Croatia)

    2013-07-01

    Full text: The Rudjer Boskovic Institute Tandem Accelerator Facility is equipped with a number of end-stations dedicated to ion beam analysis (IBA), modification of materials and nuclear physics experiments. IBA is performed at the: (1) nuclear microbeam, (2) broad-beam in-vacuum and (3) external beam end-stations. Several lBA techniques can be used simultaneously, Particle Induced X-ray Emission (PIXE) and Particle Induced Gamma-ray Emission (PIGE) at the external beam end-station, and additionally Rutherford Backscattering Spectroscopy (RBS) at the in-vacuum end-station. X-ray fluorescence (XRF) analysis is a technique complementary to PIXE. Both techniques offer high analytical potential for multi-elemental investigations and material characterization. Due to different excitation mechanisms, PIXE generally exhibits higher sensitivity for lighter elements and XRF for heavier, whereas they also have different in-sample depth sensitivities. Although they use different excitation sources, both techniques can use the same data acquisition modules. With the development of miniature, low power and lightweight X-ray tubes it is possible to incorporate an X-ray source within the IBA setup and combine the two techniques for simultaneous use. In this work, the unification of the PIXE and XRF techniques at the RBI external ion beam analysis setup has been investigated and the results are discussed. This has been done by installing a transmission miniature X-ray tube at the end-station. The tube has been properly positioned in order to irradiate the same spot on the sample as the ion beam used for PIXE/PIGE measurements. Our home made data acquisition system SPECTOR, used regularly for the IBA measurements, has been also used to acquire the XRF spectra. At first, the X-ray tube has been installed at the in-vacuum IBA station, and then to the external beam end-station. Test measurements have been carried out on various standard reference materials using both systems and the

  6. Quantitative Analysis of Various Metalloprotein Compositional Stoichiometries with Simultaneous PIXE and NRA

    Science.gov (United States)

    McCubbin, Andrew; Deyoung, Paul; Peaslee, Graham; Sibley, Megan; Warner, Joshua

    2013-04-01

    Stoichiometric characterization has been carried out on multiple metalloproteins using a combination of Ion Beam Analysis methods and a newly modified preparation technique. Particle Induced X-ray emission (PIXE) spectroscopy is a non-destructive ion beam analysis technique well suited to determine the concentrations of heavy elements. Nuclear Reaction Analysis (NRA) is a technique which measures the areal density of a thin target from scattering cross sections of 3.4 MeV protons. A combination of NRA and PIXE has been developed to provide a quantitative technique for the determination of stoichiometric metal ion ratios in metalloproteins. About one third of all proteins are metalloproteins, and most do not have well determined stoichiometric compositions for the metals they contain. Current work focuses on establishing a standard method in which to prepare protein samples. The method involves placing drops of protein solutions on aluminized polyethylene terephthalate (Mylar) and allowing them to dry. This technique has been tested for several proteins of known stoichiometry to determine cofactor content and has proven to be a reliable analysis method, accurately determining metal stoichiometry in cytochrome c, superoxide dismutase, concanavalin A, vitamin B12, and hemoglobin.

  7. A simultaneous CONCOR algorithm for the analysis of two partitioned matrices

    NARCIS (Netherlands)

    Lafosse, R; Ten Berge, J.M.F.

    2006-01-01

    A standard approach to derive underlying components from two or more data matrices, holding data from the same individuals or objects, is the (generalized) canonical correlation analysis. This technique finds components (canonical variates) with maximal sums of correlations between them. The

  8. Simultaneous control of error rates in fMRI data analysis.

    Science.gov (United States)

    Kang, Hakmook; Blume, Jeffrey; Ombao, Hernando; Badre, David

    2015-12-01

    The key idea of statistical hypothesis testing is to fix, and thereby control, the Type I error (false positive) rate across samples of any size. Multiple comparisons inflate the global (family-wise) Type I error rate and the traditional solution to maintaining control of the error rate is to increase the local (comparison-wise) Type II error (false negative) rates. However, in the analysis of human brain imaging data, the number of comparisons is so large that this solution breaks down: the local Type II error rate ends up being so large that scientifically meaningful analysis is precluded. Here we propose a novel solution to this problem: allow the Type I error rate to converge to zero along with the Type II error rate. It works because when the Type I error rate per comparison is very small, the accumulation (or global) Type I error rate is also small. This solution is achieved by employing the likelihood paradigm, which uses likelihood ratios to measure the strength of evidence on a voxel-by-voxel basis. In this paper, we provide theoretical and empirical justification for a likelihood approach to the analysis of human brain imaging data. In addition, we present extensive simulations that show the likelihood approach is viable, leading to "cleaner"-looking brain maps and operational superiority (lower average error rate). Finally, we include a case study on cognitive control related activation in the prefrontal cortex of the human brain.

  9. Simultaneous Comparison of Two Roller Compaction Techniques and Two Particle Size Analysis Methods.

    Science.gov (United States)

    Saarinen, Tuomas; Antikainen, Osmo; Yliruusi, Jouko

    2017-05-24

    A new dry granulation technique, gas-assisted roller compaction (GARC), was compared with conventional roller compaction (CRC) by manufacturing 34 granulation batches. The process variables studied were roll pressure, roll speed, and sieve size of the conical mill. The main quality attributes measured were granule size and flow characteristics. Within granulations also the real applicability of two particle size analysis techniques, sieve analysis (SA) and fast imaging technique (Flashsizer, FS), was tested. All granules obtained were acceptable. In general, the particle size of GARC granules was slightly larger than that of CRC granules. In addition, the GARC granules had better flowability. For example, the tablet weight variation of GARC granules was close to 2%, indicating good flowing and packing characteristics. The comparison of the two particle size analysis techniques showed that SA was more accurate in determining wide and bimodal size distributions while FS showed narrower and mono-modal distributions. However, both techniques gave good estimates for mean granule sizes. Overall, SA was a time-consuming but accurate technique that provided reliable information for the entire granule size distribution. By contrast, FS oversimplified the shape of the size distribution, but nevertheless yielded acceptable estimates for mean particle size. In general, FS was two to three orders of magnitude faster than SA.

  10. Simultaneous determination of aflatoxins B2 and G2 in peanuts using spectrofluorescence coupled with parallel factor analysis.

    Science.gov (United States)

    Luna, A S; Luiz, R A; Lima, I C A; Março, P H; Valderrama, P; Boqué, R; Ferré, J

    2013-05-17

    In the present study a method for the simultaneous determination of aflatoxins B2 and G2 in peanuts has been developed. The method uses second order standard addition method and excitation-emission fluorescence data together with parallel factor analysis (PARAFAC). The aflatoxin analysis was based on extraction with methanol-water and carried out using immunoaffinity clean-up. The results of PARAFAC on a set of spiked and naturally contaminated peanuts indicated that the two aflatoxins could be successfully determined. The method was validated and analytical figures of merit were obtained for both analytes. The limits of detection (LOD) were 0.05 and 0.04 μg kg(-1) for aflatoxins B2 and G2, respectively. The limits of quantification (LOQ) were 0.16 and 0.12 μg kg(-1) for aflatoxins B2 and G2, respectively. Coupling of spectrofluorimetry with PARAFAC can be considered as an alternative method for quantification of aflatoxins in the presence of unknown interferences obtained through analysis of highly complex matrix of peanuts samples at a reduced cost per analysis.

  11. Simultaneous analysis of gamma-hydroxybutyric acid and its precursors in urine using liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Wood, Michelle; Laloup, Marleen; Samyn, Nele; Morris, Michael R; de Bruijn, Ernst A; Maes, Robert A; Young, Michael S; Maes, Viviane; De Boeck, Gert

    2004-11-12

    We have developed a rapid method that enables the simultaneous analysis of gamma-hydroxybutyrate (GHB) and its precursors, i.e. gamma-butyrolactone (GBL) and 1,4-butanediol (1,4-BD) in urine. The method comprised a simple dilution of the urine sample, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Chromatographic separation was achieved using an Atlantis dC18 column, eluted with a mixture of formic acid and methanol. The method was linear from 1-80 mg/L for GHB and 1,4-BD and from 1-50 mg/L for GBL. The limit of quantification was 1 mg/L for all analytes. The procedure, which has a total analysis time (including sample preparation) of less than 12 min, was fully validated and applied to the analysis of 182 authentic urine samples; the results were correlated with a previously published GC-MS procedure and revealed a low prevalence of GHB-positive samples. Since no commercial immunoassay is available for the routine screening of GHB, this simple and rapid method should prove useful to meet the current increased demand for the measurement of GHB and its precursors.

  12. Co-transcriptomic Analysis by RNA Sequencing to Simultaneously Measure Regulated Gene Expression in Host and Bacterial Pathogen

    KAUST Repository

    Ravasi, Timothy

    2016-01-24

    Intramacrophage pathogens subvert antimicrobial defence pathways using various mechanisms, including the targeting of host TLR-mediated transcriptional responses. Conversely, TLR-inducible host defence mechanisms subject intramacrophage pathogens to stress, thus altering pathogen gene expression programs. Important biological insights can thus be gained through the analysis of gene expression changes in both the host and the pathogen during an infection. Traditionally, research methods have involved the use of qPCR, microarrays and/or RNA sequencing to identify transcriptional changes in either the host or the pathogen. Here we describe the application of RNA sequencing using samples obtained from in vitro infection assays to simultaneously quantify both host and bacterial pathogen gene expression changes, as well as general approaches that can be undertaken to interpret the RNA sequencing data that is generated. These methods can be used to provide insights into host TLR-regulated transcriptional responses to microbial challenge, as well as pathogen subversion mechanisms against such responses.

  13. Fingerprint analysis and simultaneous determination of phenolic compounds in extracts of Curculiginis Rhizoma by HPLC-diode array detector.

    Science.gov (United States)

    Bian, Qingya; Yang, Hui; Chan, Chi-On; Jin, Dengping; Mok, Daniel Kam-Wah; Chen, Sibao

    2013-01-01

    Curculiginis Rhizoma (Curculigo orchioides GAERTN.) is a well-known Chinese herbal medicine, as well as an important Rasayana drug in India. Current criteria of quality control on this herb are to quantitatively analyze single compound curculigoside, which fail to comprehensively evaluate quality of this herb. In this paper, a simple and reliable HPLC coupled with diode array detector (DAD) method was developed to evaluate the quality of Curculiginis Rhizoma through establishing chromatographic fingerprint and simultaneously quantitating four phenolic compounds, orcinol glucoside, orcinol, 2,6-dimethoxybenzoic acid and curculigoside. The fingerprint displayed eleven common peaks, and the similarity index of different samples was in a range of 0.890-0.977. Validation of the method was acceptable, with 96.03-102.82% accuracy in recovery test and inter and intra-day precisions were less than 2%. This developed method by having a combination of chromatographic fingerprint and quantitation analysis could be applied to the quality control of Curculiginis Rhizoma.

  14. Simultaneous analysis method for polar and non-polar ginsenosides in red ginseng by reversed-phase HPLC-PAD.

    Science.gov (United States)

    Lee, Sa-Im; Kwon, Ha-Jeong; Lee, Yong-Moon; Lee, Je-Hyun; Hong, Seon-Pyo

    2012-02-23

    The paper describes the development of a simultaneous determination method for polar and non-polar ginsenosides in red ginseng with a reversed-phase high-performance liquid chromatography-pulsed amperometric detection method. This method could be applied directly without any pretreatment steps and enabled the performance of highly sensitive analysis within 1h. The detection (S/N=3) and quantification (S/N=10) limits for the ginsenosides ranged 0.02-0.10 ng and 0.1-0.3 ng, respectively. The linear regression coefficients ranged 0.9975-0.9998. Intra- and inter-day precisions were <9.91%. The mean recoveries ranged 98.08-103.06%. The total amount of ginsenosides in the hairy root of red ginseng was higher than that in the main root.

  15. Simultaneous determination of nitrite and nitrate ions by air-segmented amplitude-modulated multiplexed flow analysis.

    Science.gov (United States)

    Yoshida, Haruka; Inui, Koji; Takeuchi, Masaki; Tanaka, Hideji

    2012-01-01

    The concept of amplitude-modulated multiplexed flow analysis has been extended to the simultaneous determination of multiple analytes in a sample. A sample solution containing nitrite and nitrate ions is delivered from two channels, but the flow rates are varied at different frequencies. One of the channels has a reduction column for converting nitrate ions to nitrite ions. Downstream, the absorbance of the diazo-coupling product is monitored after the merging of both solutions with a Griess reagent. The signal is analyzed by a fast Fourier transform (FFT) in real time. From the thus-obtained amplitude, a µmol dm(-3) level of the ions can be determined. The introduction of air bubbles is effective to reduce any axial dispersion, and hence to improve the sensitivity.

  16. Characterizing the Smell of Marijuana by Odor Impact of Volatile Compounds: An Application of Simultaneous Chemical and Sensory Analysis.

    Directory of Open Access Journals (Sweden)

    Somchai Rice

    Full Text Available Recent U.S. legislation permitting recreational use of marijuana in certain states brings the use of marijuana odor as probable cause for search and seizure to the forefront of forensic science, once again. This study showed the use of solid-phase microextraction with multidimensional gas chromatography--mass spectrometry and simultaneous human olfaction to characterize the total aroma of marijuana. The application of odor activity analysis offers an explanation as to why high volatile chemical concentration does not equate to most potent odor impact of a certain compound. This suggests that more attention should be focused on highly odorous compounds typically present in low concentrations, such as nonanal, decanol, o-cymene, benzaldehyde, which have more potent odor impact than previously reported marijuana headspace volatiles.

  17. Simultaneous extraction and derivatization of carbohydrates from green plant tissues for analysis by gas-liquid chromatography.

    Science.gov (United States)

    Streeter, J G; Strimbu, C E

    1998-06-01

    Simultaneous extraction and derivatization of carbohydrates was performed by mixing dry ground plant tissue with derivatization reagents in pyridine; trimethylsilyl derivatives were analyzed by gas-liquid chromatography. This "direct analysis" was compared to analysis of samples prepared by exhaustive ethanol extraction of the same ground plant tissues. Comparisons included leaf blades from apple, grape, corn, and tomato and leaf blade, petiole, stem, and pod tissues from soybean plants. Direct analysis gave superior quantification of sucrose, glucose, and fructose because of sucrose hydrolysis during ethanol extraction. Sucrose hydrolysis was highly variable among plant species and use of hot ethanol at the first extraction step reduced sucrose hydrolysis but did not always abolish it. Sucrose hydrolysis was probably due to the activity of hydrolytic enzymes in 75% ethanol at room temperature. Direct analysis was inferior for the quantification of cyclitols in the fibrous tissues of soybean but provided acceptable results for cyclitol analysis in leaf blade tissue. When the time for extraction/reaction was extended from 40 to 60 min, some improvement in recovery of cyclitols was observed, but recovery remained 10 to 20% below that obtained with exhaustive ethanol extraction. Mannitol was vacuum infiltrated into the five types of leaf tissue and recovery averaged 100% by the direct method relative to ETOH extraction for apple, grape, corn, and soybean leaves but was only 76% for tomato leaves. Direct analysis provides very large time savings and is clearly the method of choice when the analysis of large numbers of samples of plant tissues for carbohydrate composition is required.

  18. Analysis of metabolites in plasma reveals distinct metabolic features between Dahl salt-sensitive rats and consomic SS.13(BN) rats.

    Science.gov (United States)

    Wang, Le; Hou, Entai; Wang, Zhengjun; Sun, Na; He, Liqing; Chen, Lan; Liang, Mingyu; Tian, Zhongmin

    2014-07-18

    Salt-sensitive hypertension is a major risk factor for cardiovascular disorders. Our previous proteomic study revealed substantial differences in several proteins between Dahl salt-sensitive (SS) rats and salt-insensitive consomic SS.13(BN) rats. Subsequent experiments indicated a role of fumarase insufficiency in the development of hypertension in SS rats. In the present study, a global metabolic profiling study was performed using gas chromatography/mass spectrometry (GC/MS) in plasma of SS rats (n=9) and SS.13(BN) rats (n=8) on 0.4% NaCl diet, designed to gain further insights into the relationship between alterations in cellular intermediary metabolism and predisposition to hypertension. Principal component analysis of the data sets revealed a clear clustering and separation of metabolic profiles between SS rats and SS.13(BN) rats. 23 differential metabolites were identified (PSS rats. Pyruvate, which connects TCA cycle and glycolysis, was also increased in SS rats. Moreover, lower activity levels of fumarase, aconitase, α-ketoglutarate dehydrogenase and succinyl-CoA synthetase were detected in the heart, liver or skeletal muscles of SS rats. The distinct metabolic features in SS and SS.13(BN) rats indicate abnormalities of TCA cycle in SS rats, which may play a role in predisposing SS rats to developing salt-sensitive hypertension.

  19. A green analytical method for the simultaneous analysis of cefixime trihydrate and ambroxol HCl based on ultraviolet derivative spectroscopy

    Directory of Open Access Journals (Sweden)

    Ceema Mathew

    2015-01-01

    Full Text Available Context: Until date, there is no reported derivative spectrophotometric method for the combination of cefixime trihydrate (CEF and ambroxol HCl (ABH. So an urgent need was felt to develop an ultraviolet (UV derivative spectroscopic method, which reduces the cost of analysis on comparing with high-performance liquid chromatography or high-performance thin layer chromatography method. Aims: To develop and validate an economical and ecofriendly derivative spectroscopic method that avoids the use of organic solvents for simultaneous quantification of both the drugs. Materials and Methods: A simple method based on the derivative spectrophotometric method at zero crossing wavelengths has been developed for the simultaneous quantification of CEF and ABH. As the method depends on hydrotropic dissolution, 0.1N urea is used as the solvent, and it yields an economical and ecofriendly method. Two wavelengths 253 nm (zero crossing point (ZCP for CEF and 306 nm (ZCP for ABH were selected for the quantification of ABH and CEF respectively. Results: The first derivative amplitude-concentration plots were linear over the range of 5-35 μg/ml and 3-10.5 μg/ml with detection limits of 0.187 and 0.0937 μg/ml and quantification limits of 0.625 and 0.312 μg/ml for CEF and ABH respectively. The percentage recovery was within the range between 99.05% and 102%. The % relative standard deviation for precision and accuracy of the method was found to be <2%. Conclusion: The proposed method was found to be simple, accurate and precise and can be successfully applied to the routine quality control analysis of studied drugs in their tablet formulations.

  20. Coherent source and connectivity analysis on simultaneously measured EEG and MEG data during isometric contraction.

    Science.gov (United States)

    Muthuraman, M; Hellriegel, H; Hoogenboom, N; Anwar, A R; Mideksa, K G; Krause, H; Schnitzler, A; Raethjen, J; Deuschl, G

    2014-01-01

    The most well-known non-invasive electric and magnetic field measurement modalities are the electroencephalography (EEG) and magnetoencephalography (MEG). The first aim of the study was to implement the recently developed realistic head model which uses an integrative approach for both the modalities. The second aim of this study was to find the network of coherent sources and the modes of interactions within this network during isometric contraction (ISC) at (15-30 Hz) in healthy subjects. The third aim was to test the effective connectivity revealed by both the modalities analyzing them separately and combined. The Welch periodogram method was used to estimate the coherence spectrum between the EEG and the electromyography (EMG) signals followed by the realistic head modelling and source analysis method dynamic imaging of coherent sources (DICS) to find the network of coherent sources at the individual peak frequency within the beta band in healthy subjects. The last step was to identify the effective connectivity between the identified sources using the renormalized partial directed coherence method. The cortical and sub-cortical network comprised of the primary sensory motor cortex (PSMC), secondary motor area (SMA), and the cerebellum (C). The cortical and sub-cortical network responsible for the isometric contraction was similar in both the modalities when analysing them separately and combined. The SNR was not significantly different between the two modalities separately and combined. However, the coherence values were significantly higher in the combined modality in comparison to each of the modality separately. The effective connectivity analysis revealed plausible additional connections in the combined modality analysis.

  1. Simultaneous chromatographic analysis of photoinitiators and amine synergists in food contact materials.

    Science.gov (United States)

    Lago, M A; Rodríguez-Bernaldo de Quirós, A; Sendón, R; Bustos, J; Santillana, M I; Paseiro, P

    2014-07-01

    Photoinitiators (PIs) are components of UV-cured inks widely used in printing of food packaging. These substances can migrate into food and may be a hazard to human health. High-performance liquid chromatography with diode-array detection (HPLC-DAD) has been used for analysis of PIs and amine synergists in food packaging. Analysis was performed with a Kromasil C18 column (250 mm × 3.2 mm i.d., 5 μm particle size) with a binary mobile phase gradient prepared from acetonitrile and Milli-Q water. The flow rate was 0.5 mL min(-1). The method enables separation of fourteen PIs and amine synergists in a single run. The method was validated for linearity, repeatability, and limits of detection and quantification. Excellent sensitivity (LODs ≤ 1.56 μg dm(2)) and appropriate repeatability (RSD (n = 10) <0.9%) were achieved. Different types of food packaging material including plastic films, cardboard, and cans were analyzed and PIs were detected in 47% of the samples tested (n = 17). Positive samples were confirmed by use of LC-MS-MS in positive electrospray ionization (ESI) mode.

  2. Functional evaluation of peripheral nerve regeneration in the rat : walking track analysis

    NARCIS (Netherlands)

    Varejao, ASP; Meek, MF; Patricio, JAB; Cabrita, AMS

    2001-01-01

    The experimental model of choice for many peripheral nerve investigators is the rat. Walking track analysis is a useful tool in the evaluation of functional peripheral nerve recovery in the rat. This quantitative method of analyzing hind limbs performance by examining footprints, known as the sciati

  3. Simultaneous determination of a novel calcium entry blocker, monatepil maleate, and its metabolites in rat plasma by means of solid-phase extraction and reversed-phase liquid chromatography with electrochemical detection.

    Science.gov (United States)

    Kurono, M; Suzuki, K; Yoshida, K; Naruto, S

    1997-02-21

    A reversed-phase LC method with electrochemical detection is described for the simultaneous determination of monatepil maleate (AJ-2615, AJ), a novel calcium entry blocker, and its three S-oxidized metabolites in plasma. These compounds were extracted from plasma by solid-phase extraction and injected onto an ODS column. The determination limit in plasma (0.5 ml) was 10 ng/ml for AJ and 5 ng/ml for the three metabolites. The method was applied to the determination of AJ and the metabolites in rat plasma samples.

  4. Derivation and transcriptional profiling analysis of pluripotent stem cell lines from rat blastocysts

    Institute of Scientific and Technical Information of China (English)

    Chunliang Li; Ying Yang; Junjie Gu; Yu Ma; Ying Jin

    2009-01-01

    Embryonic stem (ES) cells are derived from blastocyst-stage embryos. Their unique properties of self-renewal and pluripotency make them an attractive tool for basic research and a potential cell resource for therapy. ES cells of mouse and human have been successfully generated and applied in a wide range of research. However, no genuine ES cell lines have been obtained from rat to date. In this study, we identified pluripotent cells in early rat embryos using specific antibodies against markers of pluripotent stem cells. Subsequently, by modifying the culture medium for rat blastocysts, we derived pluripotent rat ES-llke cell lines, which expressed pluripotency markers and formed embryoid bodies (EBs) in vitro. Importantly, these rat ES-like cells were able to produce teratomas. Both EBs and teratomas contained tissues from all three embryonic germ layers, in addition, from the rat ES-like cells, we derived a rat primitive endoderm (PrE) cell line. Furthermore, we conducted transcriptional profiling of the rat ES-like cells and identified the unique molecular signature of the rat pluripotent stem cells. Our analysis demonstrates that multiple signaling pathways, including the BMP, Activin and roTOR pathways, may be involved in keeping the rat ES-like cells in an undifferentiated state. The cell lines and information obtained in this study will accelerate our understanding of the molecular regulation underlying pluripotency and guide us in the appropriate manipulation of ES cells from a particular species.

  5. NMR Based Cerebrum Metabonomic Analysis Reveals Simultaneous Interconnected Changes during Chick Embryo Incubation.

    Science.gov (United States)

    Feng, Yue; Zhu, Hang; Zhang, Xu; Wang, Xuxia; Xu, Fuqiang; Tang, Huiru; Ye, Chaohui; Liu, Maili

    2015-01-01

    To find out if content changes of the major functional cerebrum metabolites are interconnected and formed a network during the brain development, we obtained high-resolution magic-angle-spinning (HR-MAS) 1H NMR spectra of cerebrum tissues of chick embryo aged from incubation day 10 to 20, and postnatal day 1, and analyzed the data with principal component analysis (PCA). Within the examined time window, 26 biological important molecules were identified and 12 of them changed their relative concentration significantly in a time-dependent manner. These metabolites are generally belonged to three categories, neurotransmitters, nutrition sources, and neuronal or glial markers. The relative concentration changes of the metabolites were interconnected among/between the categories, and, more interestingly, associated with the number and size of Nissl-positive neurons. These results provided valuable biochemical and neurochemical information to understand the development of the embryonic brain.

  6. Production of furfural from pentosan-rich biomass: analysis of process parameters during simultaneous furfural stripping.

    Science.gov (United States)

    Agirrezabal-Telleria, I; Gandarias, I; Arias, P L

    2013-09-01

    Among the furan-based compounds, furfural (FUR) shows interesting properties as building-block or industrial solvent. It is produced from pentosan-rich biomass via xylose cyclodehydration. The current FUR production makes use of homogeneous catalysts and excessive amounts of steam. The development of greener furfural production and separation techniques implies the use of heterogeneous catalysts and innovative separation processes. This work deals with the conversion of corncobs as xylose source to be dehydrated to furfural. The results reveal differences between the use of direct corncob hydrolysis and dehydration to furfural and the prehydrolysis and dehydration procedures. Moreover, this work focuses on an economical analysis of the main process parameters during N2-stripping and its economical comparison to the current steam-stripping process. The results show a considerable reduction of the annual utility costs due to use of recyclable nitrogen and the reduction of the furfural purification stages.

  7. Simultaneous Analysis of Recurrent Jovian Electron Increases and Galactic Cosmic Ray Decreases

    CERN Document Server

    Kühl, P; Dunzlaff, P; Fichtner, H; Gieseler, J; Gómez-Herrero, R; Heber, B; Klassen, A; Kleimann, J; Kopp, A; Potgieter, M; Scherer, K; Strauss, R D

    2013-01-01

    The transport environment for particles in the heliosphere, e.g. galactic cosmic rays (GCRs) and MeV electrons (including those originating from Jupiters magnetosphere), is defined by the solar wind flow and the structure of the embedded heliospheric magnetic field. Solar wind structures, such as co-rotating interaction regions (CIR), can result in periodically modulation of both particles species. A detailed analysis of this recurrent Jovian electron events and galactic cosmic ray decreases measured by SOHO EPHIN is presented here, showing clearly a change of phase between both phenomena during the cause of the years 2007 and 2008. This effect can be explained by the change of difference in heliolongitude between the Earth and Jupiter, which is of central importance for the propagation of Jovian electrons. Furthermore, the data can be ordered such that the 27-day Jovian electron variation vanishes in the sector which does not connect the Earth with Jupiter magnetically using observed solar wind speeds.

  8. Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis

    Directory of Open Access Journals (Sweden)

    Karlovsky Petr

    2006-01-01

    Full Text Available Abstract Background Fusarium head blight (FHB is a disease of cereal crops, which has a severe impact on wheat and barley production worldwide. Apart from reducing the yield and impairing grain quality, FHB leads to contamination of grain with toxic secondary metabolites (mycotoxins, which pose a health risk to humans and livestock. The Fusarium species primarily involved in FHB are F. graminearum and F. culmorum. A key prerequisite for a reduction in the incidence of FHB is an understanding of its epidemiology. Results We describe a duplex-PCR-based method for the simultaneous detection of F. culmorum and F. graminearum in plant material. Species-specific PCR products are identified by melting curve analysis performed in a real-time thermocycler in the presence of the fluorescent dye SYBR Green I. In contrast to multiplex real-time PCR assays, the method does not use doubly labeled hybridization probes. Conclusion PCR with product differentiation by melting curve analysis offers a cost-effective means of qualitative analysis for the presence of F. culmorum and F. graminearum in plant material. This method is particularly suitable for epidemiological studies involving a large number of samples.

  9. Gemini NorthNIRI Spectra of Pluto and Charon: Simultaneous Analysis of the Surface and Atmosphere

    Science.gov (United States)

    Cook, Jason C.; Cruikshank, Dale P.; Young, Leslie A.

    2014-01-01

    94035We report on our analysis of blended Pluto and Charon spectra over the wavelength range 1.4 to 2.5 m as obtained by the NIRI instrument on Gemini North on June 25-28, 2004. The data have a resolving power () around 1500 and a SNR around 200 per pixel. The observed blended spectra are compared to models that combine absorption from the solid ice on the surface using Hapke theory, and absorption from the gaseous atmosphere. We assume the spectrum is a combination of several spatially separate spectral units: a CH4-rich ice unit, a volatile unit (an intimate mixture of N2, CH4 and CO), and a Charon unit (H2O, ammonia hydrate and kaolinite). We test for the presence of hydrocarbons (i.e. C2H6) and nitriles (i.e. HCN) and examine cases where additional ices are present as either pure separate spatial units, mixed with the CH4-rich unit or part of the volatile unit. We conclude that 2-4 of Plutos surface is covered with pure-C2H6 and our identification of C2H6 is significantly strengthened when absorption due to gaseous CH4 is included. The inclusion of Plutos atmosphere demonstrates that low-resolution, high-SNR observations are capable of detecting Plutos atmosphere during a time when Plutos atmosphere may have been undergoing rapid changes (1988-2002) and no high-resolution spectra were obtained. In particular, we identify features at 1.665 and 2.317 m as the Q-branch of the 23 and 3+4 bands of gaseous CH4, respectively. The later band is also evident in many previously published spectra of Pluto. Our analysis finds it is unnecessary to include 13CO to explain the depth of the 2.405 m, which has been previously suggested to be a spectral blended with C2H6, but we cannot definitively rule out its presence. Funding for this work (Cook) has been provided by a NASA-PATM grant.

  10. Simultaneous measurement of deuterium distribution and impurities by emission angle analysis of deuteron induced reaction products

    CERN Document Server

    Kubota, N; Furuyama, Y; Kitamura, A

    2002-01-01

    A novel analytical method of light element distribution in a thin film is presented. The method is based on the deuteron-induced nuclear reaction. The emission angle of the lighter product detected coincidentally with the heavier product is analyzed to deduce the depth distribution of the target atoms, while the conventional energy analysis is applied for impurities, the distributions of which are not of primary interest. Results of proof-of-principle experiments using the D(d,p)t reaction for a deuterated polyethylene (C sub 2 D sub 4) film are described. The depth resolution is evaluated to be 0.66+-0.07 mu m for 400 keV deuteron incidence in the C sub 2 D sub 4 film. Factors limiting the resolution are discussed, and possible improvement even down to several tens of nm is concluded. The present method is applicable for microanalysis of some light elements other than deuterium contained in a film with thickness of several mu m which cannot be reached by conventional heavy ion elastic recoil detection using ...

  11. Measuring Electrolyte Impedance and Noise Simultaneously by Triangular Waveform Voltage and Principal Component Analysis.

    Science.gov (United States)

    Xu, Shanzhi; Wang, Peng; Dong, Yonggui

    2016-04-22

    In order to measure the impedance variation process in electrolyte solutions, a method of triangular waveform voltage excitation is investigated together with principal component analysis (PCA). Using triangular waveform voltage as the excitation signal, the response current during one duty cycle is sampled to construct a measurement vector. The measurement matrix is then constructed by the measurement vectors obtained from different measurements. After being processed by PCA, the changing information of solution impedance is contained in the loading vectors while the response current and noise information is contained in the score vectors. The measurement results of impedance variation by the proposed signal processing method are independent of the equivalent impedance model. The noise-induced problems encountered during equivalent impedance calculation are therefore avoided, and the real-time variation information of noise in the electrode-electrolyte interface can be extracted at the same time. Planar-interdigitated electrodes are experimentally tested for monitoring the KCl concentration variation process. Experimental results indicate that the measured impedance variation curve reflects the changing process of solution conductivity, and the amplitude distribution of the noise during one duty cycle can be utilized to analyze the contact conditions of the electrode and electrolyte interface.

  12. Measuring Electrolyte Impedance and Noise Simultaneously by Triangular Waveform Voltage and Principal Component Analysis

    Directory of Open Access Journals (Sweden)

    Shanzhi Xu

    2016-04-01

    Full Text Available In order to measure the impedance variation process in electrolyte solutions, a method of triangular waveform voltage excitation is investigated together with principal component analysis (PCA. Using triangular waveform voltage as the excitation signal, the response current during one duty cycle is sampled to construct a measurement vector. The measurement matrix is then constructed by the measurement vectors obtained from different measurements. After being processed by PCA, the changing information of solution impedance is contained in the loading vectors while the response current and noise information is contained in the score vectors. The measurement results of impedance variation by the proposed signal processing method are independent of the equivalent impedance model. The noise-induced problems encountered during equivalent impedance calculation are therefore avoided, and the real-time variation information of noise in the electrode-electrolyte interface can be extracted at the same time. Planar-interdigitated electrodes are experimentally tested for monitoring the KCl concentration variation process. Experimental results indicate that the measured impedance variation curve reflects the changing process of solution conductivity, and the amplitude distribution of the noise during one duty cycle can be utilized to analyze the contact conditions of the electrode and electrolyte interface.

  13. Real-time analysis of laser beams by simultaneous imaging on a single camera chip

    Science.gov (United States)

    Piehler, S.; Boley, M.; Abdou Ahmed, M.; Graf, T.

    2015-03-01

    The fundamental parameters of a laser beam, such as the exact position and size of the focus or the beam quality factor M² yield vital information both for laser developers and end-users. However, each of these parameters can significantly change on a short time scale due to thermally induced effects in the processing optics or in the laser source itself, leading to process instabilities and non-reproducible results. In order to monitor the transient behavior of these effects, we have developed a camera-based measurement system, which enables full laser beam characterization in online. A novel monolithic beam splitter has been designed which generates a 2D array of images on a single camera chip, each of which corresponds to an intensity cross section of the beam along the propagation axis separated by a well-defined spacing. Thus, using the full area of the camera chip, a large number of measurement planes is achieved, leading to a measurement range sufficient for a full beam characterization conforming to ISO 11146 for a broad range of beam parameters of the incoming beam. The exact beam diameters in each plane are derived by calculation of the 2nd order intensity moments of the individual intensity slices. The processing time needed to carry out both the background filtering and the image processing operations for the full analysis of a single camera image is in the range of a few milliseconds. Hence, the measurement frequency of our system is mainly limited by the frame-rate of the camera.

  14. Simultaneous measurement of contact angle and surface tension using axisymmetric drop-shape analysis-no apex (ADSA-NA).

    Science.gov (United States)

    Kalantarian, A; David, R; Chen, J; Neumann, A W

    2011-04-05

    Axisymmetric drop-shape analysis-no apex (ADSA-NA) is a recent drop-shape method that allows the simultaneous measurement of contact angles and surface tensions of drop configurations without an apex (i.e., a sessile drop with a capillary protruding into the drop). Although ADSA-NA significantly enhanced the accuracy of contact angle and surface tension measurements compared to that of original ADSA using a drop with an apex, it is still not as accurate as a surface tension measurement using a pendant drop suspended from a holder. In this article, the computational and experimental aspects of ADSA-NA were scrutinized to improve the accuracy of the simultaneous measurement of surface tensions and contact angles. It was found that the results are relatively insensitive to different optimization methods and edge detectors. The precision of contact angle measurement was enhanced by improving the location of the contact points of the liquid meniscus with the solid substrate to subpixel resolution. To optimize the experimental design, the capillary was replaced with an inverted sharp-edged pedestal, or holder, to control the drop height and to ensure the axisymmetry of the drops. It was shown that the drop height is the most important experimental parameter affecting the accuracy of the surface tension measurement, and larger drop heights yield lower surface tension errors. It is suggested that a minimum nondimensional drop height (drop height divided by capillary length) of 1.7 is required to reach an error of less than 0.2 mJ/m(2) for the measured surface tension. As an example, the surface tension of water was measured to be 72.46 ± 0.04 at 24 °C by ADSA-NA, compared to 72.39 ± 0.01 mJ/m(2) obtained with pendant drop experiments.

  15. Motor function in Parkinson's disease and supranuclear palsy: simultaneous factor analysis of a clinical scale in several populations

    Directory of Open Access Journals (Sweden)

    Cubo Esther

    2006-06-01

    Full Text Available Abstract Background In order to better understand the similarities and differences in the motor behaviour of different groups of patients, their scores on the Motor Examination section of the Unified Parkinson's Disease Rating Scale (UPDRS were analysed simultaneously. The three groups consisted, respectively, of patients with Parkinson's disease (PD on medication, patients with Parkinson's disease withdrawn from anti-parkinsonian medication for at least 12 hours, and patients diagnosed with a specific Parkinsonism syndrome: Progressive Supranuclear Palsy (PSP. Methods A total of 669 consecutively sampled patients from three separate hospital-based clinics participated (294 PD on medication; 200 PD off medication: 175 PSP. The Motor Examination section of the UPDRS was administered by neurologists at the three participating clinics. The patient scores on each item were recorded. To assess similarities and differences among the components of the UPDRS in these samples, we performed simultaneous or multigroup factor analysis on the covariance matrices of the three groups. In addition, it was investigated whether a single model for the Motor Examination section of the UPDRS could be developed which would be valid for all three groups at the same time. Results A single six-dimensional factor solution was found that fitted all groups, although this was not straightforward due to differences between the tremor-at-rest variables. The factors were identified as Tremor-at-rest, Postural Tremor, Axial Dysfunctioning, Rigidity, Left Bradykinesia and Right Bradykinesia. The analysis also pointed to a somewhat lower lateralization in bradykinesia for PSP patients. The groups differed in intensity of motor impairment, especially with respect to Tremor-at-Rest, but the overall relationships between the variables were shared by the three groups. In addition, apart from the common factor structure evidence of differences in body part-specific and motor

  16. The effects of hypotension on differences between the results of simultaneous venous and arterial blood gas analysis

    Directory of Open Access Journals (Sweden)

    Farhad Shirani

    2011-01-01

    Full Text Available Background: Analysis of venous blood gas (VBG can represent arterial blood gas (ABG analysis in patients with various diseases. The effects of hypotension on differences between the results of simultaneous venous and arterial blood gas analyses were reviewed. Methods: This observational, cross-sectional study was conducted from March to October 2010 in emergency depart-ments of two university hospitals in Tehran (Iran on consecutive adult patients for whom ABG had been indicated for diagnosis/treatment. Arterial and peripheral venous bloods were simultaneously sampled with blood pressure measure-ment. The VBG-ABG amount of difference regarding pH, HCO 3 , PCO 2 , PO 2 , SO 2 , and Base Excess (BE was com-pared between those with and without hypotension. Results: During the study, 192 patients (51.6 ± 23.6 years, 67.7% males were entered into the hypotension (n = 78 and normotensive groups (n = 114. The average VBG-ABG amount of difference (95% limits of agreement in the hypotension versus normotensive group were -0.030 (-0.09 to 0.03 vs. -0.016 (-0.1 to 0.068 for pH (p = 0.01, 1.79 (-1.91 to 5.49 vs. 1.32 (-1.94 to 4.58 mEq/L for HCO 3 (p = 0.032, 2.69 (-20.43 to 25.81 vs. 2.03 (-7.75 to 11.81 mmHg for PCO 2 (p = 0.295, -35.97 (-130.17 to 58.23 vs. -32.65 (-104.79 to 39.49 mmHg for PO 2 (p = 0.293, -18.58 (-14.66 to 51.82 vs. -9.06 (-31.28 to 13.16 percent (p < 0.001 for SO 2 , and 0.25 (-3.73 to 4.23 vs. 0.79 (-2.51 to 4.09 for BE (p = 0.036. Conclusions: Hypotensive status is associated with an increase in the amount of difference between VBG and ABG analysis regarding pH, HCO 3 , and BE, though the amount of increase does not seem to be clinically important. Studying the precise effects of replacing ABG with VBG on the clinical decision-making and the following outcomes is worth-while.

  17. The effects of hypotension on differences between the results of simultaneous venous and arterial blood gas analysis.

    Science.gov (United States)

    Shirani, Farhad; Salehi, Reza; Naini, Afsoon Emami; Azizkhani, Reza; Gholamrezaei, Ali

    2011-02-01

    Analysis of venous blood gas (VBG) can represent arterial blood gas (ABG) analysis in patients with various diseases. The effects of hypotension on differences between the results of simultaneous venous and arterial blood gas analyses were reviewed. This observational, cross-sectional study was conducted from March to October 2010 in emergency departments of two university hospitals in Tehran (Iran) on consecutive adult patients for whom ABG had been indicated for diagnosis/treatment. Arterial and peripheral venous bloods were simultaneously sampled with blood pressure measurement. The VBG-ABG amount of difference regarding pH, HCO(3), PCO(2), PO(2), SO(2), and Base Excess (BE) was compared between those with and without hypotension. During the study, 192 patients (51.6 ± 23.6 years, 67.7% males) were entered into the hypotension (n = 78) and normotensive groups (n = 114). The average VBG-ABG amount of difference (95% limits of agreement) in the hypotension versus normotensive group were -0.030 (-0.09 to 0.03) vs. -0.016 (-0.1 to 0.068) for pH (p = 0.01), 1.79 (-1.91 to 5.49) vs. 1.32 (-1.94 to 4.58) mEq/L for HCO(3) (p = 0.032), 2.69 (-20.43 to 25.81) vs. 2.03 (-7.75 to 11.81) mmHg for PCO(2) (p = 0.295), -35.97 (-130.17 to 58.23) vs. -32.65 (-104.79 to 39.49) mmHg for PO(2) (p = 0.293), -18.58 (-14.66 to 51.82) vs. -9.06 (-31.28 to 13.16) percent (p analysis regarding pH, HCO(3), and BE, though the amount of increase does not seem to be clinically important. Studying the precise effects of replacing ABG with VBG on the clinical decision-making and the following outcomes is worth-while.

  18. Simultaneous speciation analysis of chromate, molybdate, tungstate and vanadate in welding fume alkaline extracts by HPLC-ICP-MS.

    Science.gov (United States)

    Ščančar, Janez; Berlinger, Balázs; Thomassen, Yngvar; Milačič, Radmila

    2015-09-01

    A novel analytical procedure was developed for the simultaneous speciation analysis of chromate, molybdate, tungstate and vanadate by anion-exchange high performance liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (HPLC-ICP-MS). Linear gradient elution from 100% water to 100% 0.7 M NaCl was applied for chromatographic separation of metal species. In standard aqueous solution at neutral pH molybdate, tungstate and vanadate exist in several aqueous species, while chromate is present as a single CrO4(2-) species. Consequently, only chromate can be separated from this solution in a sharp chromatographic peak. For obtaining sharp chromatographic peaks for molybdate, tungstate and vanadate, the pH of aqueous standard solutions was raised to 12. At highly alkaline conditions single CrO4(2-), MoO4(2-) and WO4(2-) are present and were eluted in sharp chromatographic peaks, while VO4(3-) species, which predominates at pH 12 was eluted in slightly broaden peak. In a mixture of aqueous standard solutions (pH 12) chromate, molybdate, tungstate and vanadate were eluted at retention times from 380 to 420 s, 320 to 370 s, 300 to 350 s and 240 to 360 s, respectively. Eluted species were simultaneously detected on-line by ICP-MS recording m/z 52, 95, 182 and 51. The developed procedure was successfully applied to the analysis of leachable concentrations of chromate, molybdate, tungstate and vanadate in alkaline extracts (2% NaOH+3% Na2CO3) of manual metal arc (MMA) welding fumes loaded on filters. Good repeatability and reproducibility of measurement (RSD±3.0%) for the investigated species were obtained in both aqueous standard solutions (pH 12) and in alkaline extracts of welding fumes. Low limits of detection (LODs) were found for chromate (0.02 ng Cr mL(-1)), molybdate (0.1 ng Mo mL(-1)), tungstate (0.1 ng W mL(-1)) and vanadate (0.2 ng V mL(-1)). The accuracy of analytical procedure for the determination of chromate was checked by analysis of

  19. Simultaneous analysis and peroxynitrite-scavenging activity of galloylated flavonoid glycosides and ellagic acid in Euphorbia supina.

    Science.gov (United States)

    Nugroho, Agung; Rhim, Tae-Jin; Choi, Moo-Young; Choi, Jae Sue; Kim, Youn-Chul; Kim, Min-Sun; Park, Hee-Juhn

    2014-07-01

    The herbs of Euphorbia supina (Euphorbiaceae) have been used to treat hemorrhage, chronic bronchitis, hepatitis, jaundice, diarrhea, gastritis, and hemorrhoids as a medicinal herb. This work is aimed to qualitatively and quantitatively analyze the polyphenols with peroxynitrite-scavenging activities. The eight compounds: gallic acid, methyl gallate, avicularin, astragalin, juglanin, isoquercitrin 6″-gallate, astragalin 6″-gallate, and ellagic acid, were isolated from E. supina and used for HPLC analysis and peroxynitrite (ONOO(-))-scavenging assay. Simultaneous analysis of the eight compounds was performed on MeOH extract and its fractions. The contents in MeOH extract and peroxynitrite-scavenging activities of the dimer of gallic acid, ellagic acid (15.64 mg/g; IC50 0.89 μM), and two galloylated flavonoid glycosides, astragalin 6″-gallate (13.72 mg/g; IC50 1.43 μM) and isoquercitrin 6″-gallate (16.99 mg/g; IC50 1.75 μM), were high, compared to other compounds. The legendary uses of E. supina could be attributed to the high content of polyphenols, particularly ellagic acid, isoquercitrin 6″-gallate, and astragalin 6″-gallate as active principles.

  20. Simultaneous transcriptome analysis of Colletotrichum gloeosporioides and tomato fruit pathosystem reveals novel fungal pathogenicity and fruit defense strategies.

    Science.gov (United States)

    Alkan, Noam; Friedlander, Gilgi; Ment, Dana; Prusky, Dov; Fluhr, Robert

    2015-01-01

    The fungus Colletotrichum gloeosporioides breaches the fruit cuticle but remains quiescent until fruit ripening signals a switch to necrotrophy, culminating in devastating anthracnose disease. There is a need to understand the distinct fungal arms strategy and the simultaneous fruit response. Transcriptome analysis of fungal-fruit interactions was carried out concurrently in the appressoria, quiescent and necrotrophic stages. Conidia germinating on unripe fruit cuticle showed stage-specific transcription that was accompanied by massive fruit defense responses. The subsequent quiescent stage showed the development of dendritic-like structures and swollen hyphae within the fruit epidermis. The quiescent fungal transcriptome was characterized by activation of chromatin remodeling genes and unsuspected environmental alkalization. Fruit response was portrayed by continued highly integrated massive up-regulation of defense genes. During cuticle infection of green or ripe fruit, fungi recapitulate the same developmental stages but with differing quiescent time spans. The necrotrophic stage showed a dramatic shift in fungal metabolism and up-regulation of pathogenicity factors. Fruit response to necrotrophy showed activation of the salicylic acid pathway, climaxing in cell death. Transcriptome analysis of C. gloeosporioides infection of fruit reveals its distinct stage-specific lifestyle and the concurrent changing fruit response, deepening our perception of the unfolding fungal-fruit arms and defenses race.

  1. Automated method for simultaneous lead and strontium isotopic analysis applied to rainwater samples and airborne particulate filters (PM10).

    Science.gov (United States)

    Beltrán, Blanca; Avivar, Jessica; Mola, Montserrat; Ferrer, Laura; Cerdà, Víctor; Leal, Luz O

    2013-09-03

    A new automated, sensitive, and fast system for the simultaneous online isolation and preconcentration of lead and strontium by sorption on a microcolumn packed with Sr-resin using an inductively coupled plasma mass spectrometry (ICP-MS) detector was developed, hyphenating lab-on-valve (LOV) and multisyringe flow injection analysis (MSFIA). Pb and Sr are directly retained on the sorbent column and eluted with a solution of 0.05 mol L(-1) ammonium oxalate. The detection limits achieved were 0.04 ng for lead and 0.03 ng for strontium. Mass calibration curves were used since the proposed system allows the use of different sample volumes for preconcentration. Mass linear working ranges were between 0.13 and 50 ng and 0.1 and 50 ng for lead and strontium, respectively. The repeatability of the method, expressed as RSD, was 2.1% and 2.7% for Pb and Sr, respectively. Environmental samples such as rainwater and airborne particulate (PM10) filters as well as a certified reference material SLRS-4 (river water) were satisfactorily analyzed obtaining recoveries between 90 and 110% for both elements. The main features of the LOV-MSFIA-ICP-MS system proposed are the capability to renew solid phase extraction at will in a fully automated way, the remarkable stability of the column which can be reused up to 160 times, and the potential to perform isotopic analysis.

  2. Simultaneous analysis of micro-RNA and DNA for determining the body fluid origin of DNA profiles.

    Science.gov (United States)

    van der Meer, Donny; Uchimoto, Mari L; Williams, Graham

    2013-07-01

    Micro-RNAs (miRNAs) can be specifically expressed in forensically relevant body fluids such as blood or saliva. The aim of the study was to develop a simultaneous extraction and analysis protocol that allows for the acquisition of a DNA profile and the identity of the body fluid using a single process. DNA and micro-RNA were extracted from blood and saliva before undergoing a cDNA synthesis step by using stem-loop reverse transcription PCR. The resulting extracts containing DNA and cDNA synthesized from body fluid-specific miRNA markers then underwent standard STR analysis using a modified ABI AmpFℓSTR(®) NGM SElect™ kit. In all samples, a full DNA profile was obtained along with additional peaks corresponding to the miRNA marker targeted. In all cases, blood samples profiled exhibited a peak indicating the presence of the blood-specific miRNA marker and the saliva sample profiled exhibited a peak indicating the presence of the saliva-specific miRNA marker. © 2013 American Academy of Forensic Sciences.

  3. ASE extraction method for simultaneous carbon and nitrogen stable isotope analysis in soft tissues of aquatic organisms

    Energy Technology Data Exchange (ETDEWEB)

    Bodin, Nathalie [ISM/LPTC, UMR 5255 CNRS, Universite de Bordeaux 1, 351 Cours de la Liberation, 33405 Talence (France); CRH, UMR 212 EME, Institut de Recherche et de Developpement, Av Jean Monnet BP171, 34203 Sete (France); Budzinski, Helene, E-mail: h.budzinski@ism.u-bordeaux1.fr [ISM/LPTC, UMR 5255 CNRS, Universite de Bordeaux 1, 351 Cours de la Liberation, 33405 Talence (France); Le Menach, Karyn; Tapie, Nathalie [ISM/LPTC, UMR 5255 CNRS, Universite de Bordeaux 1, 351 Cours de la Liberation, 33405 Talence (France)

    2009-06-08

    Since lipids are depleted in {sup 13}C relative to proteins and carbohydrates, variations in lipid composition among species and within individuals significantly influence {delta}{sup 13}C and may result in misleading ecological interpretations. Whereas lipid extraction before IRMS analysis constitutes a way of stable isotope result lipid-normalisation, such a procedure was given up because of the un-controlled effects of the methods used (i.e., 'Bligh and Dyer', Soxhlet, etc.) on {delta}{sup 15}N. The aim of this work was to develop a simple, rapid and efficient lipid extraction method allowing for simultaneous C and N stable isotope analysis in the biological soft tissues of aquatic organisms. The goal was to be free from the lipid influence on {delta}{sup 13}C values without interfering with {delta}{sup 15}N values. For that purpose, the modern automated pressurized liquid extraction technique ASE (accelerated solvent extraction) was selected. Eel muscles representative of a broad range of fat contents were extracted via ASE by using different semi-polar solvents (100% dichloromethane and 80% n-hexane/20% acetone) and by operating at different temperature (ambient temperature and 100 deg. C) and pressure (750 and 1900 psi) conditions. The results were discussed in terms of lipid extraction efficiency as well as {delta}{sup 13}C and {delta}{sup 15}N variability.

  4. Fractal analysis of alveolarization in hyperoxia-induced rat models of bronchopulmonary dysplasia.

    Science.gov (United States)

    Porzionato, Andrea; Guidolin, Diego; Macchi, Veronica; Sarasin, Gloria; Grisafi, Davide; Tortorella, Cinzia; Dedja, Arben; Zaramella, Patrizia; De Caro, Raffaele

    2016-04-01

    No papers are available about potentiality of fractal analysis in quantitative assessment of alveolarization in bronchopulmonary dysplasia (BPD). Thus, we here performed a comparative analysis between fractal [fractal dimension (D) and lacunarity] and stereological [mean linear intercept (Lm), total volume of alveolar air spaces, total number of alveoli, mean alveolar volume, total volume and surface area of alveolar septa, and mean alveolar septal thickness] parameters in experimental hyperoxia-induced models of BPD. At birth, rats were distributed between the following groups: 1) rats raised in ambient air for 2 wk; 2) rats exposed to 60% oxygen for 2 wk; 3) rats raised in normoxia for 6 wk; and 4) rats exposed to 60% hyperoxia for 2 wk and to room air for further 4 wk. Normoxic 6-wk rats showed increased D and decreased lacunarity with respect to normoxic 2-wk rats, together with changes in all stereological parameters except for mean alveolar volume. Hyperoxia-exposed 2-wk rats showed significant changes only in total number of alveoli, mean alveolar volume, and lacunarity with respect to equal-in-age normoxic rats. In the comparison between 6-wk rats, the hyperoxia-exposed group showed decreased D and increased lacunarity, together with changes in all stereological parameters except for septal thickness. Analysis of receiver operating characteristic curves showed a comparable discriminatory power of D, lacunarity, and total number of alveoli; Lm and mean alveolar volume were less discriminative. D and lacunarity did not show significant changes when different segmentation thresholds were applied, suggesting that the fractal approach may be fit to automatic image analysis. Copyright © 2016 the American Physiological Society.

  5. SNP and haplotype mapping for genetic analysis in the rat.

    NARCIS (Netherlands)

    Saar, K.; Beck, A.; Bihoreau, M.; Birney, E.; Brocklebank, D.; Chen, Y.; Cuppen, E.; Demonchy, S.; Dopazo, J.; Flicek, P.; Foglio, M.; Fujiyama, A.; Gut, I.G.; Gauguier, D.; Guigo, R.; Guryev, V.; Heinig, M.; Hummel, O.; Jahn, N.; Klages, S.; Kren, V.; Kube, M.; Kuhl, H.; Kuramoto, T.; Kuroki, Y.; Lechner, D.; Lee, Y.A.; Lopez-Bigas, N.; Lathrop, G.M.; Mashimo, T.; Medina, I.; Mott, R.; Patone, G.; Perrier-Cornet, J.A.; Platzer, M.; Pravenec, M.; Reinhardt, R.; Sakaki, Y.; Schilhabel, M.; Schulz, H.; Serikawa, T.; Shikhagaie, M.; Tatsumoto, S.; Taudien, S.; Toyoda, A.; Voigt, B.; Zelenika, D.; Zimdahl, H.; Hubner, N.

    2008-01-01

    The laboratory rat is one of the most extensively studied model organisms. Inbred laboratory rat strains originated from limited Rattus norvegicus founder populations, and the inherited genetic variation provides an excellent resource for the correlation of genotype to phenotype. Here, we report a s

  6. SNP and haplotype mapping for genetic analysis in the rat

    NARCIS (Netherlands)

    Saar, Kathrin; Beck, Alfred; Bihoreau, Marie-Thérèse; Birney, Ewan; Brocklebank, Denise; Chen, Yuan; Cuppen, Edwin; Demonchy, Stephanie; Dopazo, Joaquin; Flicek, Paul; Foglio, Mario; Fujiyama, Asao; Gut, Ivo G; Gauguier, Dominique; Guigo, Roderic; Guryev, Victor; Heinig, Matthias; Hummel, Oliver; Jahn, Niels; Klages, Sven; Kren, Vladimir; Kube, Michael; Kuhl, Heiner; Kuramoto, Takashi; Kuroki, Yoko; Lechner, Doris; Lee, Young-Ae; Lopez-Bigas, Nuria; Lathrop, G Mark; Mashimo, Tomoji; Medina, Ignacio; Mott, Richard; Patone, Giannino; Perrier-Cornet, Jeanne-Antide; Platzer, Matthias; Pravenec, Michal; Reinhardt, Richard; Sakaki, Yoshiyuki; Schilhabel, Markus; Schulz, Herbert; Serikawa, Tadao; Shikhagaie, Medya; Tatsumoto, Shouji; Taudien, Stefan; Toyoda, Atsushi; Voigt, Birger; Zelenika, Diana; Zimdahl, Heike; Hubner, Norbert

    2008-01-01

    The laboratory rat is one of the most extensively studied model organisms. Inbred laboratory rat strains originated from limited Rattus norvegicus founder populations, and the inherited genetic variation provides an excellent resource for the correlation of genotype to phenotype. Here, we report a s

  7. Simultaneous Determination of Eight Alkaloids in Rat Plasma by UHPLC-MS/MS after Oral Administration of Coptis deltoidea C. Y. Cheng et Hsiao and Coptis chinensis Franch

    Directory of Open Access Journals (Sweden)

    Lu Liu

    2016-07-01

    Full Text Available A ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS method was successfully developed and validated for the identification and determination of eight alkaloids: tetrahydropalmatine (A; palmatine (B; magnoflorine (C; columbamine (D; berberine (E; worenine (F; berberrubine (G and coptisine (H in rat plasma, which are the active components in Coptis deltoidea C. Y. cheng et Hsiao (CCY and Coptis chinensis Franch (CF. The chromatographic separation of analytes was successfully achieved on an Agilent SB-C18 column (1.8 µm, 150 mm × 2.1 mm using a programme with a mobile phase consisting of acetonitrile and water containing 0.3% acetic acid at a flow rate of 0.25 mL/min. The analytes were detected with a triple quadrupole tandem MS in multiple reaction monitoring (MRM mode and an electrospray ionization (ESI source in positive mode. The validated method showed good linearity over a wide concentration range (r2 > 0.991, and lower limits of quantification (LLOQ less than 1.1 ng/mL for all analytes, and matrix effects ranged from 85.2% to 106.8%. The mean extraction recoveries were no less than 86.4%, and the precision and accuracy were within the acceptable limits. All analytes were proven to be stable during sample storage and analysis procedures. The method validation results demonstrated that the proposed method was sensitive, specific, and reliable, which could lay a foundation for the pharmacokinetic study of eight analytes after oral administration of CCY and CF in subsequent studies.

  8. Simultaneous determination of baicalin, oroxylin A-7-O-glucuronide and wogonoside in rat plasma by UPLC-DAD and its application in pharmacokinetics of pure baicalin, Radix Scutellariae and Yinhuang granule.

    Science.gov (United States)

    Chen, Hui; Li, Zheng; Li, Yin-jie; Wu, Xiao-wen; Wang, Shi-rui; Chen, Kai; Zheng, Xiao-xiao; Du, Qian; Tang, Dao-quan

    2015-12-01

    A novel UPLC-DAD method was developed and validated for the simultaneous determination of baicalin (baicalein-7-glucuronide, BG), oroxylin A-7-O-glucuronide (OAG) and wogonoside (WG) in rat plasma using rutin as the internal standard. Plasma samples were precipitated using acetonitrile containing 0.1% formic acid. Separation was performed on an Agilent Eclipse Plus C18 column (2.1 × 50 mm, 1.8 µm) using gradient acetonitrile and 0.2% formic acid water solution as mobile phase. The flow-rate was set at 0.4 mL/min and the eluate was detected at 275 nm. The method was linear over the ranges of 0.075-17.50, 0.050-12.60 and 0.056-14.10 µg/mL for BG, OAG and WG, respectively. The intra- and inter-day precisions were respectively <4.8% and 6.4%. All of the limits of detection of three analytes in rat plasma were 0.01 µg/mL, whereas the limits of quantification were, respectively, 0.035, 0.025 and, 0.025 µg/mL. This assay has been successfully applied to pharmacokinetics of BG, OAG and WG in rats after oral administration of Yinhuang granule (YHG) and comparative pharmacokinetics of BG in rats following oral administration of the pure BG, Radix Scutellariae (RS) or YHG. We speculate that some co-existing ingredients in RS or YHG may increase the absorption and elimination of BG in rat. This work may be helpful for the quality control of Yinhuang granule.

  9. Simultaneous determination of l-tetrahydropalmatine and its active metabolites in rat plasma by a sensitive ultra-high-performance liquid chromatography with tandem mass spectrometry method and its application in a pharmacokinetic study.

    Science.gov (United States)

    Wang, Weihui; Liu, Jing; Zhao, Xiaoning; Peng, Yan; Wang, Nannan; Lee, David Y W; Dai, Ronghua

    2017-06-01

    A sensitive and reliable ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for simultaneous determination of l-tetrahydropalmatine (l-THP) and its active metabolites l-isocorypalmine (l-ICP) and L-corydalmine (l-CD) in rat plasma. The analytes were extracted by liquid-liquid extraction and separated on a Bonshell ASB C18 column (2.1 × 100 mm; 2.7 μm; Agela) using acetonitrile-formic acid aqueous as mobile phase at a flow rate of 0.2 mL/min in gradient mode. The method was validated over the concentration range of 4.00-2500 ng/mL for l-THP, 0.400-250 ng/mL for l-ICP and 1.00-625 ng/mL for l-CD. Intra- and inter-day accuracy and precision were within the acceptable limits of 0.99 for all analytes. The quantitative method was successfully applied for simultaneous determination of l-THP and its active metabolites in a pharmacokinetic study after oral administration with l-THP at a dose of 15 mg/kg to rats. Copyright © 2016 John Wiley & Sons, Ltd.

  10. Simultaneous analysis of cortisol and cortisone in saliva using XLC-MS/MS for fully automated online solid phase extraction.

    Science.gov (United States)

    Jones, Rachel L; Owen, Laura J; Adaway, Joanne E; Keevil, Brian G

    2012-01-15

    Salivary cortisol measurements are increasingly being used in the investigation of disorders of the hypothalamic-pituitary-adrenal axis. In the salivary gland, cortisol is metabolised to cortisone by the action of 11β-hydroxysteroid dehydrogenase type 2, and cortisone is partly responsible for the variable interference observed in current salivary cortisol immunoassays. The aim of this study was to validate an assay for the simultaneous analysis of salivary cortisol and cortisone using the Spark Holland Symbiosis™ in eXtraction liquid chromatography-tandem mass spectrometry (XLC-MS/MS) mode for fully automated online solid phase extraction (SPE). Saliva samples were diluted in water with the addition of internal standard (d4-cortisol and d7-cortisone). Online SPE was performed using the Spark Holland Symbiosis™ with HySphere™ C18 SPE cartridges and compounds were eluted onto a Phenomenex® C18 guard column attached to a Phenomenex® Onyx monolithic C18 column for chromatography. Mass spectrometry used the Waters® Xevo™ TQ MS in electrospray positive mode. Cortisol and cortisone eluted with their internal standards at 1.95 and 2.17 min, respectively, with a total run time of four minutes. No evidence of ion-suppression was observed. The assay was linear up to 3393 nmol/L for cortisol and 3676 nmol/L for cortisone, with lower limits of quantitation of 0.75 nmol/L and 0.50 nmol/L, respectively. Intra- and inter-assay imprecision was cortisone across three levels of internal quality control, with accuracy and recovery within accepted limits. High specificity was demonstrated following interference studies which assessed 29 structurally-related steroids at supra-physiological concentrations. We have successfully validated an assay for the simultaneous analysis of salivary cortisol and cortisone using XLC-MS/MS and fully automated online SPE. The assay benefits from increased specificity compared to immunoassay and minimal sample preparation which allows high

  11. Genetic and molecular analysis of radon-induced rat lung tumours; Analyse cytogenetique et moleculaire des tumeurs pulmonaires radon-induites chez le rat

    Energy Technology Data Exchange (ETDEWEB)

    Guilly, M.N.; Joubert, Ch.; Levalois, C.; Dano, L.; Chevillard, S. [CEA Fontenay-aux-Roses, Dept. de Radiobiologie et de Radiopathologie, 92 (France)

    2002-03-01

    We have a model of radon-induced rat lung tumours, which allow us to analyse the cytogenetic and molecular alterations of the tumours. The aim is to better understand the mechanisms of radio-induced carcinogenesis and to define if it exists a specificity of radio-induced genetic alterations as compared to the genetic alterations found in the sporadic tumours. We have started our analysis by developing global cytogenetic and molecular approaches. We have shown that some alterations are recurrent. The genes that are potentially involved are the oncogene MET and the tumour suppressor Bene p16, which are also frequently altered in human lung tumours. Simultaneously, we have focussed our analysis by targeting the search of mutation in the tumour suppressor gene TP3. We have found that 8 of 39 tumours were mutated by deletion in the coding sequence of TP53. This high frequency of deletion, which is not observed in the human p53 mutation database could constitute a signature of radio-induced alterations. On this assumption, this type of alteration should not be only found on TP53 Bene but also in other suppressor genes which are inactivated by a mutation such as p16 for example. The work we are carrying out on radio-induced tumours among humans and animals is directed to this end. (author)

  12. Current source density analysis: methods and application to simultaneously recorded field potentials of the rabbit's visual cortex.

    Science.gov (United States)

    Rappelsberger, P; Pockberger, H; Petsche, H

    1981-01-01

    This paper deals with the application of current source density (CSD) analysis to simultaneously recorded intracortical field potentials of the rabbit's visual cortex. Recordings were made with multielectrodes with either 8 contacts at distances of 300 microns, or 16 contacts at distances of 150 microns on one carrier needle. For synchronized activities, a spatial resolution of 150 microns turned out to be sufficient to record all depth-varying details of the field potentials; for seizure potentials even a spacing of 300 microns was adequate in most cases. For practical application, an appropriate spacing of the measuring points has to be chosen for a satisfactory estimation of the first and second derivatives of the field potentials. For this reason an interpolation procedure is applied to reduce the spacing from 300 microns or 150 microns electrode contact distances, respectively, and to obtain intermediate values at 75 microns distances. With this spacing satisfactory estimations of the second derivative are obtained. Theoretically, CSD analysis has to be made three-dimensionally, but under certain conditions which are discussed, a one-dimensional analysis can be applied. An unknown quantity is sigma z, the vertical conductivity. It turned out that average values obtained from different experiments are not representative and that the vertical conductivity has to be measured in every experiment. This is caused by the great individual differences of the cortices even if the same stereotactic coordinates are chosen. Therefore, in every experiment relative conductivity measurements are performed. The influence of different conductivity values within the various layers and the influence of a conductivity gradient is discussed and demonstrated by examples.

  13. Computational tool for morphological analysis of cultured neonatal rat cardiomyocytes.

    Science.gov (United States)

    Leite, Maria Ruth C R; Cestari, Idágene A; Cestari, Ismar N

    2015-08-01

    This study describes the development and evaluation of a semiautomatic myocyte edge-detector using digital image processing. The algorithm was developed in Matlab 6.0 using the SDC Morphology Toolbox. Its conceptual basis is the mathematical morphology theory together with the watershed and Euclidean distance transformations. The algorithm enables the user to select cells within an image for automatic detection of their borders and calculation of their surface areas; these areas are determined by adding the pixels within each myocyte's boundaries. The algorithm was applied to images of cultured ventricular myocytes from neonatal rats. The edge-detector allowed the identification and quantification of morphometric alterations in cultured isolated myocytes induced by 72 hours of exposure to a hypertrophic agent (50 μM phenylephrine). There was a significant increase in the mean surface area of the phenylephrine-treated cells compared with the control cells (p<;0.05), corresponding to cellular hypertrophy of approximately 50%. In conclusion, this edge-detector provides a rapid, repeatable and accurate measurement of cell surface areas in a standardized manner. Other possible applications include morphologic measurement of other types of cultured cells and analysis of time-related morphometric changes in adult cardiac myocytes.

  14. Simultaneous confirmatory analysis of different transgenic maize (zea mays) lines using multiplex polymerase chain reaction-restriction analysis and capillary gel electrophoresis with laser induced fluorescence detection.

    Science.gov (United States)

    García-Cañas, Virginia; Cifuentes, Alejandro

    2008-09-24

    A novel analytical procedure based on the combination of multiplex PCR, restriction analysis, and CGE-LIF to unambiguosly and simultaneously confirm the presence of multiple lines of genetically modified corn is proposed. This methodology is based on the amplification of event-specific DNA regions by multiplex PCR using 6-FAM-labeled primers. Subsequently, PCR products are digested by a mixture containing specific restriction endonucleases. Thus, restriction endonucleases selectively recognize DNA target sequences contained in the PCR products and cleave the double-stranded DNA at a given cleavage site. Next, the restriction digest is analyzed by CGE-LIF corroborating the length of the expected restriction fragments, confirming (or not) the existence of GMOs. For accurate size determination of the DNA fragments by CGE-LIF a special standard DNA mixture was produced in this laboratory for calibration. The suitability of this mixture for size determination of labeled DNA fragments is also demonstrated. The usefulness of the proposed methodology is demonstrated through the simultaneous detection and confirmatory analysis of samples containing 0.5% of GA21 and MON863 maize plus an endogenous gene of maize as control.

  15. Development of a validated UPLC method for simultaneous estimation of both free and entrapped (in solid lipid nanoparticles) all-trans retinoic acid and cholecalciferol (vitamin D3) and its pharmacokinetic applicability in rats.

    Science.gov (United States)

    Kumar, Manoj; Sharma, Gaurav; Singla, Dinesh; Singh, Sukhjeet; Sahwney, Sudhir; Chauhan, Anurag S; Singh, Gagandeep; Kaur, Indu Pal

    2014-03-01

    A sensitive ultra-performance liquid chromatography (UPLC) method was developed for simultaneous estimation of all-trans retinoic acid (ATRA) and cholecalciferol (vitamin D3) in rat plasma. The method was validated over the linear range of 1.0-5000ng/ml (r(2)=0.999) for both vitamins with a limit of detection of 0.5ng/ml. Chromatographic separation was achieved using liquid-liquid extraction (LLE) on an Acquity BEH RP 18 column (2.1mm×50mm, I.D. 1.7μm), with mobile phase comprising of acetonitrile:methanol:water (90:8:2, v/v/v), at a flow rate of 0.20ml/min and a total run time of 5min. Intra and inter-day variability (RSD) was ≤3.1%, and the accuracy varied between 95.4-99.9% and 95.3-101.1% respectively, for ATRA and 98.5-100.8% and 99.3-101.7%, respectively for vitamin D3. High recovery of ≥96.0% for ATRA and ≥87.80% for vitamin D3 was achieved. ATRA and vitamin D3 were stable in plasma under different storage and processing conditions. The method was applied to estimate the total drug content and entrapment efficiency of ATRA and vitamin D3 loaded solid lipid nanoparticles (SLNs). Concentration of these two agents was determined in rat plasma after simultaneous subcutaneous administration in free form or when loaded into SLNs thus establishing pharmacokinetic application of the developed procedure. Results indicated an improvement in AUC0-∞ by 5.4 times and 29.4 times for ATRA and vitamin D3, respectively, upon their incorporation into SLNs. Simultaneous administration of these two vitamins and their improved and prolonged bioavailability has scope for their use in treatment and control of tuberculosis.

  16. Histological analysis of heart after domoic acid administration in rats

    Directory of Open Access Journals (Sweden)

    Andrés Crespo Vieira

    2014-06-01

    All data were analysed with GraphPad Prism 5.0 by a one-way analysis of variance (ANOVA. Dunnett’s Multiple Comparison test was used for post hoc comparisons between control and treatment groups. Results were expressed as means ± SEM, with a P value of Zalophus californianus (Gulland et al. 2002; Zabka et al. 2009, although these effects were no studied under experimental conditions due to that they were natural intoxications. In vitro studies showed that DOM produced alterations in the metabolism of rat cardiac myoblasts (Nijjar et al. 1999; Vranyac-Tramoundanas et al. 2008, but few in vivo observations are reduced to a previous report by Vranyac-Tramoundanas (Vranyac-Tramoundanas et al. 2011, where heart lesions were seen 3 days i.p. administration. The early onset of the heart lesions, characteristic of acute damage, led us to realize the present assay, where we expected to observe some kind of heart damage or collagen alterations. The possible observed damage would be correlated with the presence of DOM by IHQ assays. The experimental dose (2.5 mg/kg and the antibody concentration (1:5000 were determined by previous experiments (Vieira et al., in press. Our previous observations showed an important presence of DOM only in hippocampus 6h and 10h after toxin i.p. administration (Vieira et al., in press. Neuropathological studies in rodents concluded that hippocampus constitutes the principal target of DOM, due to the high concentration of KA receptors in this area (Wisden 1993. In spite of the presence of GluRs in rat heart (Gill et al. 1998, the DOM affinity for these GluRs or the concentration of GluRs might not be high enough to detect DOM presence 6h, 10h or 24h after i.p. administration. The mild collagen alterations and the lack of observable damage in the first 24 h after toxin administration concluded that DOM seems to need several days in order to produce observable damage. Further experiments with long-time expositions to DOM will be done.

  17. Simultaneous Quantification of Diazepam and Dexamethasone in Plasma by High-Performance Liquid Chromatography with Tandem Mass Spectrometry and Its Application to a Pharmacokinetic Comparison between Normoxic and Hypoxic Rats

    Directory of Open Access Journals (Sweden)

    Wenwen Gong

    2015-04-01

    Full Text Available In order to investigate the pharmacokinetics of a combination of diazepam and dexamethasone under hypoxic conditions, a novel, sensitive and specific liquid chromatography with tandem mass spectrometry (LC-MS/MS method for the simultaneous determination of diazepam and dexamethasone in rat plasma was developed and validated. The chromatographic separation of analytes was successfully achieved on an XTerra® MS C18 column using a gradient elution of methanol and water containing 0.1% formic acid at a flow rate of 0.5 mL/min. This method demonstrated good linearity and no endogenous material interferences. The linear ranges were 1.0–100 ng/mL for diazepam and 2.0–200 ng/mL for dexamethasone. The intra- and inter-day precision for the two compounds in plasma were lower than 10.0%, and the accuracy was between −7.9% and 11.5%. Our method was then successfully applied in a pharmacokinetic comparison between normoxic and hypoxic rats. The results indicated that there were significant differences in the main pharmacokinetics parameters of diazepam and dexamethasone between normoxic and hypoxic rats. The results provide the important and valuable information for discovering and developing novel anti-hypoxia drug combinations, as well as a better understanding of the safety and efficacy of these drugs.

  18. An UPLC-MS/MS method for simultaneous quantitation of two coumarins and two flavonoids in rat plasma and its application to a pharmacokinetic study of Wikstroemia indica extract.

    Science.gov (United States)

    Wei, Lan; Wang, Xiaobo; Zhang, Peng; Sun, Yangyang; Jia, Lili; Zhao, Jingxin; Dong, Shikai; Sun, Lixin

    2016-01-01

    In this study, an ultra performance liquid chromatography tandem mass spectrometry method (UPLC-MS/MS) was developed for simultaneous determination of umbelliferone, apigenin, daphnoretin and genkwanin in total (free and conjugated) forms in rat plasma using psoralen as internal standard. Plasma samples were protein precipitated with acetonitrile followed by liquid-liquid extracted with ethyl acetate. Four ingredients were separated on an Acquity UPLC(®) BEH C18 column using gradient elution with the mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile, and detected by positive ion electrospray ionization (ESI) in multiple reaction monitoring mode (MRM). The method was linear for all analytes over investigated ranges with all correlation coefficients greater than 0.99. The validated lower limit of quantification was 3ng/mL for umbelliferone, 3ng/mL for apigenin, 12ng/mL for daphnoretin and 2ng/mL for genkwanin, respectively. Intra- and inter-day precisions (RSD%) were less than 15% and accuracy (RE%) ranged from -1.1% to 15%. The mean absolute extraction recoveries of analytes and IS from rat plasma were all more than 75%. The validated method was firstly and successfully applied to investigate the pharmacokinetics of four chemical ingredients after oral administration of Wikstroemia indica extract (WIE) to rats.

  19. Development and validation of high liquid performance chromatography-tandem mass spectrometry method for simultaneous determination of geniposidic acid and aucubin in rat plasma for pharmacokinetic study after oral administration of Du-zhong tea extract.

    Science.gov (United States)

    Zhang, Lin; Ma, Yu-Liang; Liu, Yang; Zu, Yuan-Gang

    2014-07-15

    A specific and sensitive high performance liquid chromatography coupled with tandem mass spectrometric (HPLC-MS/MS) method was developed and validated for the simultaneous determination of geniposidic acid and aucubin in rat plasma after oral administration of Du-zhong tea extract. The plasma samples were pretreated by protein precipitation with methanol and the chromatographic separation was performed on a Hypersil C18 column (4.6 mm×250 mm, 5 μm), using a gradient mobile phase system of water-methanol (0.05% formic acid). The detection was accomplished by multiple-reaction monitoring (MRM) scanning via electrospray ionization source operating in the negative ionization mode. The linear range was 1-1,000 ng/mL for geniposidic acid and 0.2-200 ng/mL for aucubin, respectively. The accuracy (relative error, R.E.%) were between -5.40 and 5.00%, while the intra-day and inter-day precisions were less than 7.95 and 7.87% for the two analytes, respectively. The method was fully validated for the sensitivity, selectivity, recovery, matrix effect and stability. Then this method was successfully applied to the pharmacokinetic study of geniposidic acid and aucubin after oral administration of Du-zhong tea extract to rats and the results indicated that this HPLC-MS/MS assay is a valuable method for the pharmacokinetic study of geniposidic acid and aucubin in rat plasma.

  20. Effects of perinatal simultaneous exposure to tributyltin (TBT) and p,p'-DDE [1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene) on male offspring of Wistar rats.

    Science.gov (United States)

    Makita, Yuji; Omura, Minoru; Ogata, Rika

    2004-03-12

    p,p'-DDE [1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene; DDE] and tributyltin (TBT) are ubiquitous in the environment and in Japan were shown to bioaccumulate in marine products. Thus these chemicals serve as a source of contaminant in the mammalian food chain. Fetuses and neonates through maternal ingestion may be exposed to DDE and TBT. Therefore, the effects of concurrent exposure to DDE and TBT were investigated in male Wistar rat offspring of dams ingesting these two contaminants. In this study, TBT suppressed the growth and delayed eye opening. However, both growth retardation and delayed eye opening produced by TBT failed to occur in the presence of DDE. Unexpectedly, the prostate weight of male rat offspring was significantly reduced with the administration of TBT but restored in the presence of DDE. These results indicate that TBT and DDE affected the development of male rat offspring following maternal exposure, and simultaneous administration of DDE prevented some of the observed effects of TBT, especially of an antagonistic nature, through a mechanism, still to be determined.

  1. Simultaneous Determination of Four Tanshinones by UPLC-TQ/MS and Their Pharmacokinetic Application after Administration of Single Ethanol Extract of Danshen Combined with Water Extract in Normal and Adenine-Induced Chronic Renal Failure Rats

    Directory of Open Access Journals (Sweden)

    Hong-Die Cai

    2016-11-01

    Full Text Available Salvia miltiorrhiza, one of the major traditional Chinese medicines, is commonly used and the main active ingredients—tanshinones—possess the ability to improve renal function. In this paper, the UPLC-TQ/MS method of simultaneously determining four tanshinones—tanshinone IIA, dihydrotanshinone I, tanshinone I, and cryptotanshinone—was established and applied to assess the pharmacokinetics in normal and chronic renal failure (CRF rat plasma. The pharmacokinetics of tanshinones in rats were studied after separately intragastric administration of Salvia miltiorrhiza ethanol extract (SMEE (0.65 g/kg, SMEE (0.65 g/kg combined with Salvia miltiorrhiza water extract (SMWE (1.55 g/kg. The results showed Cmax and AUC0–t of tanshinone IIA, tanshinone I, cryptotanshinone reduced by 50%~80% and CLz/F increased by 2~4 times (p < 0.05 in model group after administrated with SMEE. Nevertheless, after intragastric administration of a combination of SMWE and SMEE, the Cmax and AUC0–t of four tanshinones were upregulated and CLz/F was downregulated, which undulated similarity from the model group to the normal group with compatibility of SMEE and SMWE. These results hinted that SMWE could improve the bioavailability of tanshinones in CRF rats, which provides scientific information for further exploration the mechanism of the combination of SMWE and SMEE and offers a reference for clinical administration of Salvia miltiorrhiza.

  2. Simultaneous Quantification of Diazepam and Dexamethasone in Plasma by High-Performance Liquid Chromatography with Tandem Mass Spectrometry and Its Application to a Pharmacokinetic Comparison between Normoxic and Hypoxic Rats.

    Science.gov (United States)

    Gong, Wenwen; Liu, Shuhong; Xu, Pingxiang; Fan, Ming; Xue, Ming

    2015-04-16

    In order to investigate the pharmacokinetics of a combination of diazepam and dexamethasone under hypoxic conditions, a novel, sensitive and specific liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of diazepam and dexamethasone in rat plasma was developed and validated. The chromatographic separation of analytes was successfully achieved on an XTerra® MS C18 column using a gradient elution of methanol and water containing 0.1% formic acid at a flow rate of 0.5 mL/min. This method demonstrated good linearity and no endogenous material interferences. The linear ranges were 1.0-100 ng/mL for diazepam and 2.0-200 ng/mL for dexamethasone. The intra- and inter-day precision for the two compounds in plasma were lower than 10.0%, and the accuracy was between -7.9% and 11.5%. Our method was then successfully applied in a pharmacokinetic comparison between normoxic and hypoxic rats. The results indicated that there were significant differences in the main pharmacokinetics parameters of diazepam and dexamethasone between normoxic and hypoxic rats. The results provide the important and valuable information for discovering and developing novel anti-hypoxia drug combinations, as well as a better understanding of the safety and efficacy of these drugs.

  3. Comprehensive analysis of pharmaceutical products using simultaneous mixed-mode (ion-exchange/reversed-phase) and hydrophilic interaction liquid chromatography.

    Science.gov (United States)

    Kazarian, Artaches A; Nesterenko, Pavel N; Soisungnoen, Phimpha; Burakham, Rodjana; Srijaranai, Supalax; Paull, Brett

    2014-08-01

    Liquid chromatographic assays were developed using a mixed-mode column coupled in sequence with a hydrophilic interaction liquid chromatography column to allow the simultaneous comprehensive analysis of inorganic/organic anions and cations, active pharmaceutical ingredients, and excipients (carbohydrates). The approach utilized dual sample injection and valve-mediated column switching and was based upon a single high-performance liquid chromatography gradient pump. The separation consisted of three distinct sequential separation mechanisms, namely, (i) ion-exchange, (ii) mixed-mode interactions under an applied dual gradient (reversed-phase/ion-exchange), and (iii) hydrophilic interaction chromatography. Upon first injection, the Scherzo SS C18 column (Imtakt) provided resolution of inorganic anions and cations under isocratic conditions, followed by a dual organic/salt gradient to elute active pharmaceutical ingredients and their respective organic counterions and potential degradants. At the top of the mixed-mode gradient (high acetonitrile content), the mobile phase flow was switched to a preconditioned hydrophilic interaction liquid chromatography column, and the standard/sample was reinjected for the separation of hydrophilic carbohydrates, some of which are commonly known excipients in drug formulations. The approach afforded reproducible separation and resolution of up to 23 chemically diverse solutes in a single run. The method was applied to investigate the composition of commercial cough syrups (Robitussin®), allowing resolution and determination of inorganic ions, active pharmaceutical ingredients, excipients, and numerous well-resolved unknown peaks.

  4. Measurement of the heat of smoldering combustion in straws and stalks by means of simultaneous thermal analysis

    Energy Technology Data Exchange (ETDEWEB)

    He, Fang; Yi, Weiming; Zha, Jianwen [School of Light Industry and Agricultural Engineering, Shandong University of Technology, Zibo, Shandong 255049 (China)

    2009-01-15

    In order to investigate reaction heat of agro-stalks smoldering, wheat straw, corn stalk, cotton stalk, millet straw, sorghum stalk and sweet potato rattan powder were smoldered and pyrolyzed in a simultaneous thermal analyzer (STA). The samples were placed in a platinum crucible (5 mm x 5 mm) with a lid (with a 1 mm hole) on a high-accuracy differential scanning calorimetry-heat capacity (DSC-cp) holder in the furnace of an STA and heated from 303 to 1073 K at a heating rate of 10 K min{sup -1}. Sweeping gas with a flow rate of 25 ml min{sup -1} was air and nitrogen during smoldering and pyrolysis, respectively. Results showed that the heat emission characteristic of the smoldering process differed remarkably from the pyrolysis process. Based on the analysis of the DSC curves, oxidative polymer degradation heat and char oxidation heat were obtained from experimental data. It showed that the oxidative polymer degradation heat of the agro-stalks is more than 6.92 MJ kg{sup -1} consumed matter, higher than that of cellulose paper. And char oxidation heat is around 23 MJ kg{sup -1} consumed matter, similar to that of cellulose paper, but higher than that of cigarette. Total net heat emission of smoldering in STA was also obtained. These data can be used as reference data in analyzing smoldering of agro-stalks under similar conditions. (author)

  5. Novel implantable imaging system for enabling simultaneous multiplanar and multipoint analysis for fluorescence potentiometry in the visual cortex.

    Science.gov (United States)

    Kobayashi, Takuma; Motoyama, Mayumi; Masuda, Hiroyuki; Ohta, Yasumi; Haruta, Makito; Noda, Toshihiko; Sasagawa, Kiyotaka; Tokuda, Takashi; Tamura, Hideki; Ishikawa, Yasuyuki; Shiosaka, Sadao; Ohta, Jun

    2012-01-01

    Techniques for fast, noninvasive measurement of neuronal excitability within a broad area will be of major importance for analyzing and understanding neuronal networks and animal behavior in neuroscience field. In this research, a novel implantable imaging system for fluorescence potentiometry was developed using a complementary metal-oxide semiconductor (CMOS) technology, and its application to the analysis of cultured brain slices and the brain of a living mouse is described. A CMOS image sensor, small enough to be implanted into the brain, with light-emitting diodes and an absorbing filter was developed to enable real-time fluorescence imaging. The sensor, in conjunction with a voltage-sensitive dye, was certainly able to visualize the potential statuses of neurons and obtain physiological responses in both right and left visual cortex simultaneously by using multiple sensors for the first time. This accomplished multiplanar and multipoint measurement provides multidimensional information from different aspects. The light microsensors do not disturb the animal behavior. This implies that the imaging system can combine functional fluorescence imaging in the brain with behavioral experiments in a freely moving animal.

  6. Simultaneous high performance liquid chromatographic analysis of vitamins B1, B2 and B6 in royal jelly

    Directory of Open Access Journals (Sweden)

    Presoto Ana Elisa F

    2004-01-01

    Full Text Available Royal jelly is used as a food supplement, popularly known as rich in B vitamins. The present work has two objectives: firstly, to apply simultaneous quantitative determination by High Performance Liquid Chromatography of thiamin (vitamin B1, riboflavin (vitamin B2 and pyridoxine (vitamin B6 and secondly to compare the obtained data with the Dietary Reference Intake (DRI values. The values obtained showed no thiamin, a range from 20 to 171 ng g-1 of riboflavin and from 408 to 2 188 ng g-1 of pyridoxine in royal jelly. According to the Food and Nutrition Board (2000, the DRI of these vitamins varies from 0.2-1.4 mg for thiamin; 0.3-1.6 mg for riboflavin and 0.1-2.0 mg for pyridoxine, depending on age and sex. According to these recommendations, royal jelly is not a good source of vitamins B1, B2 or B6 as these vitamins appear only on order of ng g-1. The proposed method can be used in routine analysis for royal jelly, having the advantage of being simple, fast and reliable.

  7. Simultaneous blood flow and blood oxygenation measurements using a combination of diffuse speckle contrast analysis and near-infrared spectroscopy

    Science.gov (United States)

    Seong, Myeongsu; Phillips, Zephaniah; Mai, Phuong Minh; Yeo, Chaebeom; Song, Cheol; Lee, Kijoon; Kim, Jae Gwan

    2016-02-01

    A combined diffuse speckle contrast analysis (DSCA)-near-infrared spectroscopy (NIRS) system is proposed to simultaneously measure qualitative blood flow and blood oxygenation changes in human tissue. The system employs an optical switch to alternate two laser sources at two different wavelengths and a CCD camera to capture the speckle image. Therefore, an optical density can be measured from two wavelengths for NIRS measurements and a speckle contrast can be calculated for DSCA measurements. In order to validate the system, a flow phantom test and an arm occlusion protocol for arterial and venous occlusion were performed. Shorter exposure times (<1 ms) show a higher drop (between 50% and 66%) and recovery of 1/KS2 values after occlusion (approximately 150%), but longer exposure time (3 ms) shows more consistent hemodynamic changes. For four subjects, the 1/KS2 values dropped to an average of 82.1±4.0% during the occlusion period and the average recovery of 1/KS2 values after occlusion was 109.1±0.8%. There was also an approximately equivalent amplitude change in oxyhemoglobin (OHb) and deoxyhemoglobin (RHb) during arterial occlusion (max RHb=0.0085±0.0024 mM/DPF, min OHb=-0.0057±0.0044 mM/DPF). The sensitivity of the system makes it a suitable modality to observe qualitative hemodynamic trends during induced physiological changes.

  8. Simultaneous analysis of eight vitamin E isomers in Moringa oleifera Lam. leaves by ultra performance convergence chromatography.

    Science.gov (United States)

    Qi, Ningli; Gong, Xiao; Feng, Cuiping; Wang, Xiaoxi; Xu, Yongwei; Lin, Lijing

    2016-09-15

    A new method for simultaneous determination of eight vitamin E isomers including α-, β-, γ-, and δ-tocopherol and α-, β-, γ-, and δ-tocotrienol by ultra-performance convergence chromatography (UPC(2)) equipped with a diode array detector was reported. They were separated on a BEH 2-EP column (3.0 mm × 100 mm, 1.7 μm) using gradient elution (95:5-80:20) with a mobile phase consisted of CO2 and methanol:isopropanol (1:1, v/v), back pressure of 1800 psi, flow rate of 1.5 ml/min and detection at 294 nm. The results showed good linearity (R(2)=0.9990-0.9998) and high resolution (1.48-7.67). Limits of detection (LOD) and quantification (LOQ) ranged from 23-49 ng/L and 70-150 ng/L, respectively. Relative standard deviations (RSD) for repeatability and reproducibility were 0.62-3.16% and 0.82-3.34%, respectively. Moreover, this method was successfully applied to analysis the vitamin E isomers in Moringa oleifera leaf samples.

  9. Simultaneous analysis of multiple lipid oxidation products in vivo by liquid chromatographic-mass spectrometry (LC-MS).

    Science.gov (United States)

    Yin, Huiyong; Davis, Todd; Porter, Ned A

    2010-01-01

    Free radical-induced oxidation of polyunsaturated fatty acid (PUFAs) has been linked to a number of human diseases including atherosclerosis and neurodegenerative disorders. Oxidation of PUFAs generates hydroperoxides and cyclic peroxides that are reduced to lipid alcohol, such as hydroxyeicosatetraenoic acid (HETEs), and isoprostanes (IsoPs) respectively. The IsoPs are isomers of prostaglandins that are generated from autoxidation of arachidonic acid (C20:4). Quantification of F(2)-IsoPs has been regarded as the "gold standard" to assess oxidative stress status in various human diseases. We herein report the protocol of analyzing HETEs and F(2)-IsoPs using a triple quadrupole mass spectrometer coupled to reverse phase liquid chromatography. The selected reaction monitoring (SRM) mode selects the parent ion of interest in the first Quad (m/z 319 for HETE and m/z 353 for F(2)-IsoPs) and fragments it in the second while an ion characteristic of the analyte of interest is monitored in the third Quad. This highly selective technique permits the simultaneous analysis of multiple oxidation products such as the HETEs and F(2)-IsoPs. This LC-MS technique can be applied to study the free radical oxidation mechanism in vitro and assess the oxidative stress status in biological tissues and fluids.

  10. Phylogeny, classification and evolution of ladybird beetles (Coleoptera: Coccinellidae) based on simultaneous analysis of molecular and morphological data.

    Science.gov (United States)

    Seago, Ainsley E; Giorgi, Jose Adriano; Li, Jiahui; Slipiński, Adam

    2011-07-01

    Ladybird beetles (family Coccinellidae) are a species-rich, ecologically diverse group of substantial agricultural significance, yet have been consistently problematic to classify, with evolutionary relationships poorly understood. In order to identify major clades within Coccinellidae, evaluate the current classification system, and identify likely drivers of diversification in this polyphagous group, we conducted the first simultaneous Bayesian analysis of morphological and multi-locus molecular data for any beetle family. Addition of morphological data significantly improved phylogenetic resolution and support for early diverging lineages, thereby better resolving evolutionary relationships than either data type alone. On the basis of these results, we formally recognize the subfamilies Microweisinae and Coccinellinae sensuŚlipiński (2007). No significant support was found for the subfamilies Coccidulinae, Scymninae, Sticholotidinae, or Ortaliinae. Our phylogenetic results suggest that the evolutionary success of Coccinellidae is in large part attributable to the exploitation of ant-tended sternorrhynchan insects as a food source, enabled by the key innovation of unusual defense mechanisms in larvae. Copyright © 2011 Elsevier Inc. All rights reserved.

  11. A high-throughput, simultaneous analysis of carotenoids, chlorophylls and tocopherol using sub two micron core shell technology columns.

    Science.gov (United States)

    Chebrolu, Kranthi K; Yousef, Gad G; Park, Ryan; Tanimura, Yoshinori; Brown, Allan F

    2015-09-15

    A high-throughput, robust and reliable method for simultaneous analysis of five carotenoids, four chlorophylls and one tocopherol was developed for rapid screening large sample populations to facilitate molecular biology and plant breeding. Separation was achieved for 10 known analytes and four unknown carotenoids in a significantly reduced run time of 10min. Identity of the 10 analytes was confirmed by their UV-Vis absorption spectras. Quantification of tocopherol, carotenoids and chlorophylls was performed at 290nm, 460nm and 650nm respectively. In this report, two sub two micron particle core-shell columns, Kinetex from Phenomenex (1.7μm particle size, 12% carbon load) and Cortecs from Waters (1.6μm particle size, 6.6% carbon load) were investigated and their separation efficiencies were evaluated. The peak resolutions were >1.5 for all analytes except for chlorophyll-a' with Cortecs column. The ruggedness of this method was evaluated in two identical but separate instruments that produced CV<2 in peak retentions for nine out of 10 analytes separated.

  12. Validated LC-MS-MS Method for Simultaneous Analysis of 17 Barbiturates in Horse Plasma for Doping Control.

    Science.gov (United States)

    Liu, Ying; Uboh, Cornelius E; Li, Xiaoqing; Guan, Fuyu; You, Youwen; Maylin, George A; Zhu, Fengchang; Soma, Lawrence R

    2017-06-01

    A rapid and sensitive method for simultaneous screening, quantification and confirmation of 17 barbiturates in horse plasma using liquid chromatography-tandem mass spectrometry is described. Analytes were recovered from plasma by liquid-liquid extraction using methyl tert-butyl ether, separated on a C18 column, and analyzed in negative electrospray ionization mode. Multiple-reaction monitoring was employed for screening and quantification. Confirmation for the presence of the analytes was achieved by comparing ion intensity ratio. The ranges for limits of detection, quantification and confirmation were 0.003-1 ng/mL (S/N ≥ 3), 0.01-2.5 ng/mL and 0.02-5 ng/mL, respectively. The linear dynamic range of the method was 0.1-100 ng/mL. The precision and accuracy at 0.5, 5 and 50 ng/mL of all 17 barbiturates during intra-day assay were 1.6-8.6% and 96-106%, respectively. For inter-day assay, precision and accuracy at the same three concentrations were 2.6-8.9% and 96-106%, respectively. Analysis of all 17 analytes was completed within 7 min. Thus, the present method is fast, simple, sensitive and reproducibly reliable. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  13. Stability indicating simplified HPLC method for simultaneous analysis of resveratrol and quercetin in nanoparticles and human plasma.

    Science.gov (United States)

    Kumar, Sandeep; Lather, Viney; Pandita, Deepti

    2016-04-15

    Resveratrol and quercetin are well-known polyphenolic compounds present in common foods, which have demonstrated enormous potential in the treatment of a wide variety of diseases. Owing to their exciting synergistic potential and combination delivery applications, we developed a simple and rapid RP-HPLC method based on isosbestic point detection. The separation was carried out on phenomenex Synergi 4μ Hydro-RP 80A column using methanol: acetonitrile (ACN): 0.1% phosphoric acid (60:10:30) as mobile phase. The method was able to quantify nanograms of analytes simultaneously on a single wavelength (269 nm), making it highly sensitive, rapid as well as economical. Additionally, forced degradation studies of resveratrol and quercetin were established and the method's applicability was evaluated on PLGA nanoparticles and human plasma. The analytes peaks were found to be well resolved in the presence of degradation products and excipients. The simplicity of the developed method potentializes its suitability for routine in vitro and in vivo analysis of resveratrol and quercetin.

  14. Source analysis of median nerve stimulated somatosensory evoked potentials and fields using simultaneously measured EEG and MEG signals.

    Science.gov (United States)

    Mideksa, K G; Hellriegel, H; Hoogenboom, N; Krause, H; Schnitzler, A; Deuschl, G; Raethjen, J; Heute, U; Muthuraman, M

    2012-01-01

    The sources of somatosensory evoked potentials (SEPs) and fields (SEFs), which is a standard paradigm, is investigated using multichannel EEG and MEG simultaneous recordings. The hypothesis that SEP & SEF sources are generated in the posterior bank of the central sulcus is tested, and analyses are compared based on EEG only, MEG only, bandpass filtered MEG, and both combined. To locate the sources, the forward problem is first solved by using the boundary-element method for realistic head models and by using a locally-fitted-sphere approach for averaged head models consisting of a set of connected volumes, typically representing the skull, scalp, and brain. The location of each dipole is then estimated using fixed MUSIC and current-density-reconstruction (CDR) algorithms. For both analyses, the results demonstrate that the band-pass filtered MEG can localize the sources accurately at the desired region as compared to only EEG and unfiltered MEG. For CDR analysis, it looks like MEG affects EEG during the combined analyses. The MUSIC algorithm gives better results than CDR, and when comparing the two head models, the averaged and the realistic head models showed the same result.

  15. Multi-modal causality analysis of eyes-open and eyes-closed data from simultaneously recorded EEG and MEG.

    Science.gov (United States)

    Anwar, Abdul Rauf; Mideska, Kidist Gebremariam; Hellriegel, Helge; Hoogenboom, Nienke; Krause, Holger; Schnitzler, Alfons; Deuschl, Günther; Raethjen, Jan; Heute, Ulrich; Muthuraman, Muthuraman

    2014-01-01

    Owing to the recent advances in multi-modal data analysis, the aim of the present study was to analyze the functional network of the brain which remained the same during the eyes-open (EO) and eyes-closed (EC) resting task. The simultaneously recorded electroencephalogram (EEG) and magnetoencephalogram (MEG) were used for this study, recorded from five distinct cortical regions of the brain. We focused on the 'alpha' functional network, corresponding to the individual peak frequency in the alpha band. The total data set of 120 seconds was divided into three segments of 18 seconds each, taken from start, middle, and end of the recording. This segmentation allowed us to analyze the evolution of the underlying functional network. The method of time-resolved partial directed coherence (tPDC) was used to assess the causality. This method allowed us to focus on the individual peak frequency in the 'alpha' band (7-13 Hz). Because of the significantly higher power in the recorded EEG in comparison to MEG, at the individual peak frequency of the alpha band, results rely only on EEG. The MEG was used only for comparison. Our results show that different regions of the brain start to `disconnect' from one another over the course of time. The driving signals, along with the feedback signals between different cortical regions start to recede over time. This shows that, with the course of rest, brain regions reduce communication with each another.

  16. STING Millennium: a web-based suite of programs for comprehensive and simultaneous analysis of protein structure and sequence

    Science.gov (United States)

    Neshich, Goran; Togawa, Roberto C.; Mancini, Adauto L.; Kuser, Paula R.; Yamagishi, Michel E. B.; Pappas, Georgios; Torres, Wellington V.; Campos, Tharsis Fonseca e; Ferreira, Leonardo L.; Luna, Fabio M.; Oliveira, Adilton G.; Miura, Ronald T.; Inoue, Marcus K.; Horita, Luiz G.; de Souza, Dimas F.; Dominiquini, Fabiana; Álvaro, Alexandre; Lima, Cleber S.; Ogawa, Fabio O.; Gomes, Gabriel B.; Palandrani, Juliana F.; dos Santos, Gabriela F.; de Freitas, Esther M.; Mattiuz, Amanda R.; Costa, Ivan C.; de Almeida, Celso L.; Souza, Savio; Baudet, Christian; Higa, Roberto H.

    2003-01-01

    STING Millennium Suite (SMS) is a new web-based suite of programs and databases providing visualization and a complex analysis of molecular sequence and structure for the data deposited at the Protein Data Bank (PDB). SMS operates with a collection of both publicly available data (PDB, HSSP, Prosite) and its own data (contacts, interface contacts, surface accessibility). Biologists find SMS useful because it provides a variety of algorithms and validated data, wrapped-up in a user friendly web interface. Using SMS it is now possible to analyze sequence to structure relationships, the quality of the structure, nature and volume of atomic contacts of intra and inter chain type, relative conservation of amino acids at the specific sequence position based on multiple sequence alignment, indications of folding essential residue (FER) based on the relationship of the residue conservation to the intra-chain contacts and Cα–Cα and Cβ–Cβ distance geometry. Specific emphasis in SMS is given to interface forming residues (IFR)—amino acids that define the interactive portion of the protein surfaces. SMS may simultaneously display and analyze previously superimposed structures. PDB updates trigger SMS updates in a synchronized fashion. SMS is freely accessible for public data at http://www.cbi.cnptia.embrapa.br, http://mirrors.rcsb.org/SMS and http://trantor.bioc.columbia.edu/SMS. PMID:12824333

  17. Rapid detection and simultaneous antibiotic susceptibility analysis of Yersinia pestis directly from clinical specimens by use of reporter phage.

    Science.gov (United States)

    Vandamm, J P; Rajanna, C; Sharp, N J; Molineux, I J; Schofield, D A

    2014-08-01

    Yersinia pestis is a tier 1 agent due to its contagious pneumopathogenicity, extremely rapid progression, and high mortality rate. As the disease is usually fatal without appropriate therapy, rapid detection from clinical matrices is critical to patient outcomes. We previously engineered the diagnostic phage ΦA1122 with luxAB to create a "light-tagged" reporter phage. ΦA1122::luxAB rapidly detects Y. pestis in pure culture and human serum by transducing a bioluminescent signal response. In this report, we assessed the analytical specificity of the reporter phage and investigated diagnostic utility (detection and antibiotic susceptibility analysis) directly from spiked whole blood. The bioreporter displayed 100% (n = 59) inclusivity for Y. pestis and consistent intraspecific signal transduction levels. False positives were not obtained from species typically associated with bacteremia or those relevant to plague diagnosis. However, some non-pestis Yersinia strains and Enterobacteriaceae did elicit signals, albeit at highly attenuated transduction levels. Diagnostic performance was assayed in simple broth-enriched blood samples and standard aerobic culture bottles. In blood, antibiotics facilitated simultaneous antimicrobial susceptibility profiling. Consequently, the reporter phage demonstrated rapid detection and antibiotic susceptibility profiling directly from clinical samples, features that may improve patient prognosis during plague outbreaks.

  18. Simultaneous speciation analysis of inorganic nitrogen with the use of ion chromatography in highly salinated environmental samples.

    Science.gov (United States)

    Kurzyca, Iwona; Niedzielski, Przemyslaw; Frankowski, Marcin

    2016-09-01

    We present the development of a method for the simultaneous determination of inorganic nitrogen species in oxidized (NO2 (-) , NO3 (-) ) and reduced (NH4 (+) ) forms using ion chromatography with diode-array detection (205, 208, and 425 nm, respectively). The oxidized forms were determined directly after the separation in the anion exchanger, while the reduced form was determined in the column hold-up time after derivatization with the Nessler reagent. The use of an appropriate modifier (Seignette reagent) and mobile phase (NaCl) enabled the determination of inorganic nitrogen species in highly salinated environmental samples (water, sediments). Moreover, low detection limits were obtained of 0.04 mg/L for NH4 (+) and 0.006 and 0.005 mg/L for NO2 (-) and NO3 (-) , respectively. The analysis of environmental samples indicated NH4 (+) contents of up to 1161 ± 47 mg/kg and NO3 (-) of up to 148 ± 6 mg/kg for sediment samples, as well as the NH4 (+) concentrations of up to 0.98 ± 0.10 mg/L, NO2 (-) of up to 24 ± 1 mg/L and NO3 (-) of up to 20 ± 1 mg/L for water samples.

  19. Feature analysis for correlation studies of simultaneous EEG-fMRI data: A proof of concept for neurofeedback approaches.

    Science.gov (United States)

    Simoes, Simões; Lima, João; Direito, Bruno; Castelhano, João; Ferreira, Carlos; Carvalho, Paulo; Castelo-Branco, Miguel

    2015-01-01

    The identification and interpretation of facial expressions is an important feature of social cognition. This characteristic is often impaired in various neurodevelopmental disorders. Recent therapeutic approaches to intervene in social communication impairments include neurofeedback (NF). In this study, we present a NF real-time functional Magnetic Resonance Imaging (rt-fMRI), combined with electroencephalography (EEG) to train social communication skills. In this sense, we defined the right Superior Temporal Sulcus as our target region-of-interest. To analyze the correlation between the fMRI regions of interest and the EEG data, we transposed the sources located at the nearest cortical location to the target region. We extracted a set of 75 features from EEG segments and performed a correlation analysis with the brain activations extracted from rt-fMRI in the right pSTS region. The finding of significant correlations of simultaneously measured signals in distinct modalities (EEG and fMRI) is promising. Future studies should address whether the observed correlation levels between local brain activity and scalp measures are sufficient to implement NF approaches.

  20. Simultaneous determination of 76As, 122Sb and 153Sm in Chinese medicinal herbs by epithermal neutron activation analysis.

    Science.gov (United States)

    Chen, Chien-Yi

    2009-01-01

    Optimal conditions for the simultaneous determination of As, Sb and Sm in Chinese medicinal herbs using epithermal neutron activation analysis were investigated. The minimum detectable concentrations of 76As, 122Sb and 153Sm in lichen and medicinal herbs depended on the weight of the irradiated sample, and irradiation and decay durations. Optimal conditions were obtained by wrapping the irradiated target with 3.2 mm borated polyethylene neutron filters, which were adopted to screen the original reactor fission neutrons and to reduce the background activities of 38Cl, 24Na and 42K. Twelve medicinal herbs, commonly consumed by Taiwanese children as a diuretic treatment, were analysed since trace elements, such as As and Sb, in these herbs may be toxic when consumed in sufficiently large quantities over a long period. Various amounts of medicinal herbs, standardised powder, lichen and tomato leaves were weighed, packed into polyethylene bags, irradiated and counted under different conditions. The results indicated that about 350 mg of lichen irradiated for 24 h and counted for 20 min following a 30-60 h decay period was optimal for irradiation in a 10(11)n/cm s epithermal neutron flux. The implications of the content of the studied elements in Chinese medicinal herbs are discussed.

  1. CLONING AND ANALYSIS OF THE GENOMIC DNA SEQUENCE OF AUGMENTER OF LIVER REGENERATION FROM RAT

    Institute of Scientific and Technical Information of China (English)

    董菁; 成军; 王勤环; 施双双; 王刚; 斯崇文

    2002-01-01

    Objective.To search for genomic DNA sequence of the augmenter of liver regeneration (ALR) of rat.Methods.Polymerase chain reaction (PCR) with specific primers was used to amplify the sequence from the rat genome.Results.A piece of genomic DNA sequence and a piece of pseudogene of rat ALR were identified.The lengths of the gene and pseudogene are 1508 bp and 442 bp,respectively.The ALR gene of rat includes 3 exons and 2 introns.The 442 bp DNA sequence may represent a pseudogene or a ALR related peptide.Predicted amino acid sequence analysis showed that there were 14 different amino acid residues between the gene and pseudogene.ALR related peptide is 84 amino acid residues in length and relates closely to ALR protein.Conclusion.There might be a multigene family of ALR in rat.

  2. Origins of albino and hooded rats: implications from molecular genetic analysis across modern laboratory rat strains.

    Directory of Open Access Journals (Sweden)

    Takashi Kuramoto

    Full Text Available Albino and hooded (or piebald rats are one of the most frequently used laboratory animals for the past 150 years. Despite this fact, the origin of the albino mutation as well as the genetic basis of the hooded phenotype remained unclear. Recently, the albino mutation has been identified as the Arg299His missense mutation in the Tyrosinase gene and the hooded (H locus has been mapped to the ∼460-kb region in which only the Kit gene exists. Here, we surveyed 172 laboratory rat strains for the albino mutation and the hooded (h mutation that we identified by positional cloning approach to investigate possible genetic roots and relationships of albino and hooded rats. All of 117 existing laboratory albino rats shared the same albino missense mutation, indicating they had only one single ancestor. Genetic fine mapping followed by de novo sequencing of BAC inserts covering the H locus revealed that an endogenous retrovirus (ERV element was inserted into the first intron of the Kit gene where the hooded allele maps. A solitary long terminal repeat (LTR was found at the same position to the ERV insertion in another allele of the H locus, which causes the so called Irish (h(i phenotype. The ERV and the solitary LTR insertions were completely associated with the hooded and Irish coat patterns, respectively, across all colored rat strains examined. Interestingly, all 117 albino rat strains shared the ERV insertion without any exception, which strongly suggests that the albino mutation had originally occurred in hooded rats.

  3. Within-subject joint independent component analysis of simultaneous fMRI/ERP in an auditory oddball paradigm.

    Science.gov (United States)

    Mangalathu-Arumana, J; Beardsley, S A; Liebenthal, E

    2012-05-01

    The integration of event-related potential (ERP) and functional magnetic resonance imaging (fMRI) can contribute to characterizing neural networks with high temporal and spatial resolution. This research aimed to determine the sensitivity and limitations of applying joint independent component analysis (jICA) within-subjects, for ERP and fMRI data collected simultaneously in a parametric auditory frequency oddball paradigm. In a group of 20 subjects, an increase in ERP peak amplitude ranging 1-8 μV in the time window of the P300 (350-700 ms), and a correlated increase in fMRI signal in a network of regions including the right superior temporal and supramarginal gyri, was observed with the increase in deviant frequency difference. JICA of the same ERP and fMRI group data revealed activity in a similar network, albeit with stronger amplitude and larger extent. In addition, activity in the left pre- and post-central gyri, likely associated with right hand somato-motor response, was observed only with the jICA approach. Within-subject, the jICA approach revealed significantly stronger and more extensive activity in the brain regions associated with the auditory P300 than the P300 linear regression analysis. The results suggest that with the incorporation of spatial and temporal information from both imaging modalities, jICA may be a more sensitive method for extracting common sources of activity between ERP and fMRI. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Simultaneous analysis of some club drugs in whole blood using solid phase extraction and gas chromatography-mass spectrometry.

    Science.gov (United States)

    Castro, André L; Tarelho, Sónia; Silvestre, Armando; Teixeira, Helena M

    2012-02-01

    The use of psychoactive substances to improve social relations and increase body energy, in Rave Culture, has raised many legal and health public concerns, both for illicit trade and consumption. Therefore, forensic toxicology plays an important role in this area, mainly linked to the detection and quantitation of these substances, both in vivo and in post-mortem samples. In fact, at the moment, forensic sciences have been under public authorities' scrutiny and critical look, due to the increasing attention of the media and public opinion, always applying for the use of scientific knowledge to help solving forensic cases. However, forensic toxicology results are only reliable to solve legal cases if all the analytical methodologies used are appropriately validated. In this work, a methodology for the extraction and analysis of 7-aminoflunitrazepam, buprenorphine, flunitrazepam, ketamine, methadone, phencyclidine (PCP) and d-propoxyphene was developed for whole blood samples, with solid phase extraction (SPE), using OASIS(®) MCX SPE columns, and gas chromatography coupled to mass spectrometry. The procedure presented here proved to be reliable, specific, selective and sensitive, with good LODs and LOQs and good precision.The adoption of a SPE procedure with an automatic SPE extraction device, allowed an increased level of automation in sample treatment, being contemporarily less time-consuming, increasing productiveness, and allowing good recovery and appropriate selectivity being, also, simple and reproducible. The simultaneous detection and quantitation of all compounds by the same extraction and detection methodology is crucial and has a great potential for forensic toxicology and clinical analysis.

  5. Simultaneous analysis of dendritic spine density, morphology and excitatory glutamate receptors during neuron maturation in vitro by quantitative immunocytochemistry.

    Science.gov (United States)

    Nwabuisi-Heath, Evelyn; LaDu, Mary Jo; Yu, Chunjiang

    2012-06-15

    Alterations in the density and morphology of dendritic spines are characteristic of multiple cognitive disorders. Elucidating the molecular mechanisms underlying spine alterations are facilitated by the use of experimental and analytical methods that permit concurrent evaluation of changes in spine density, morphology and composition. Here, an automated and quantitative immunocytochemical method for the simultaneous analysis of changes in the density and morphology of spines and excitatory glutamate receptors was established to analyze neuron maturation, in vitro. In neurons of long-term neuron-glia co-cultures, spine density as measured by drebrin cluster fluorescence, increased from DIV (days in vitro)10 to DIV18 (formation phase), remained stable from DIV18 to DIV21 (maintenance phase), and decreased from DIV21 to DIV26 (loss phase). The densities of spine-localized NMDAR and AMPAR clusters followed a similar trend. Spine head sizes as measured by the fluorescence intensities of drebrin clusters increased from DIV10 to DIV21 and decreased from DIV21 to DIV26. Changes in the densities of NR1-only, GluR2-only, and NR1+GluR2 spines were measured by the colocalizations of NR1 and GluR2 clusters with drebrin clusters. The densities of NR1-only spines remained stable from the maintenance to the loss phases, while GluR2-only and NR1+GluR2 spines decreased during the loss phase, thus suggesting GluR2 loss as a proximal molecular event that may underlie spine alterations during neuron maturation. This study demonstrates a sensitive and quantitative immunocytochemical method for the concurrent analysis of changes in spine density, morphology and composition, a valuable tool for determining molecular events involved in dendritic spine alterations. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Simultaneous monitoring of protein adsorption at the solid-liquid interface from sessile solution droplets by ellipsometry and axisymmetric drop shape analysis by profile

    NARCIS (Netherlands)

    Wormeester, H; Busscher, HK

    1999-01-01

    In this paper two in situ techniques are combined to simultaneously examine protein adsorption at the solid-liquid interface from sessile solution droplets. With axisymmetric drop shape analysis by profile (ADSA-P) the change in solid-liquid interfacial tension is determined, while ellipsometry is e

  7. Simultaneous monitoring of protein adsorption at the solid–liquid interface from sessile solution droplets by ellipsometry and axisymmetric drop shape analysis by profile

    NARCIS (Netherlands)

    Noordmans, Jaap; Wormeester, Herbert; Busscher, Henk J.

    1999-01-01

    In this paper two in situ techniques are combined to simultaneously examine protein adsorption at the solid–liquid interface from sessile solution droplets. With axisymmetric drop shape analysis by profile (ADSA-P) the change in solid–liquid interfacial tension is determined, while ellipsometry is e

  8. Simultaneous screening analysis of 3-methyl-quinoxaline-2-carboxylic acid and quinoxaline-2-carboxylic acid residues in edible animal tissues by a competitive indirect immunoassay

    Science.gov (United States)

    Immunoassays contribute greatly to veterinary drug residue analysis and food safety, but there are no reported immunoassays on simultaneously detecting MQCA and QCA, the marker residues for carbadox and olaquindox. It is extremely difficult to produce broad-specificity antibodies that bind both res...

  9. The influence of aging on the metabolism of simultaneously administered hexobarbital enantiomers and antipyrine before and after phenobarbital induction in male rats: A longitudinal study

    NARCIS (Netherlands)

    Groen, K.; Breimer, D.D.; Jansen, E.J.; Bezooijen, C.F.A. van

    1994-01-01

    The influence of aging on the metabolism of antipyrine (AP) and hexobarbital enantiomers (R-HB and S-HB) with and without phenobarbital (PB) induction was investigated in a longitudinal study in rats aged 6, 12, 24 and 30 months. The metabolic clearances of AP (Cl(m AP)), R-HB (Cl(m R-HB)) and S-HB

  10. Simultaneous mutation detection of three homoeologous genes in wheat by High Resolution Melting analysis and Mutation Surveyor®

    Directory of Open Access Journals (Sweden)

    Vincent Kate

    2009-12-01

    Full Text Available Abstract Background TILLING (Targeting Induced Local Lesions IN Genomes is a powerful tool for reverse genetics, combining traditional chemical mutagenesis with high-throughput PCR-based mutation detection to discover induced mutations that alter protein function. The most popular mutation detection method for TILLING is a mismatch cleavage assay using the endonuclease CelI. For this method, locus-specific PCR is essential. Most wheat genes are present as three similar sequences with high homology in exons and low homology in introns. Locus-specific primers can usually be designed in introns. However, it is sometimes difficult to design locus-specific PCR primers in a conserved region with high homology among the three homoeologous genes, or in a gene lacking introns, or if information on introns is not available. Here we describe a mutation detection method which combines High Resolution Melting (HRM analysis of mixed PCR amplicons containing three homoeologous gene fragments and sequence analysis using Mutation Surveyor® software, aimed at simultaneous detection of mutations in three homoeologous genes. Results We demonstrate that High Resolution Melting (HRM analysis can be used in mutation scans in mixed PCR amplicons containing three homoeologous gene fragments. Combining HRM scanning with sequence analysis using Mutation Surveyor® is sensitive enough to detect a single nucleotide mutation in the heterozygous state in a mixed PCR amplicon containing three homoeoloci. The method was tested and validated in an EMS (ethylmethane sulfonate-treated wheat TILLING population, screening mutations in the carboxyl terminal domain of the Starch Synthase II (SSII gene. Selected identified mutations of interest can be further analysed by cloning to confirm the mutation and determine the genomic origin of the mutation. Conclusion Polyploidy is common in plants. Conserved regions of a gene often represent functional domains and have high sequence

  11. Simultaneous quantitative measurement and automated analysis of mitochondrial morphology, mass, potential, and motility in living human skin fibroblasts.

    NARCIS (Netherlands)

    Koopman, W.J.H.; Visch, H.J.; Smeitink, J.A.M.; Willems, P.H.G.M.

    2006-01-01

    BACKGROUND: Understanding the interdependence of mitochondrial and cellular functioning in health and disease requires detailed knowledge about the coupling between mitochondrial structure, motility, and function. Currently, no rapid approach is available for simultaneous quantification of these par

  12. Serum 5'nucleotidase activity in rats: a method for automated analysis and criteria for interpretation.

    Science.gov (United States)

    Carakostas, Michael C.; Power, Richard J.; Banerjee, Asit K.

    1990-01-01

    A manual kit for determining serum 5'nucleotidase (5'NT, EC 3.1.3.5) activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within-run C.V. = 2.14%; between-run C.V. = 5.5%). A comparison of the new automated method with a manual and semi-automated method gave regression statistics of y = 1.18X -3.66 (Sy. x = 4.54), and y = 0.733X + 1.97 (Sy. x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9-week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.

  13. Simultaneous determination of 18 D-amino acids in rat plasma by an ultrahigh-performance liquid chromatography-tandem mass spectrometry method: application to explore the potential relationship between Alzheimer's disease and D-amino acid level alterations.

    Science.gov (United States)

    Xing, Yuping; Li, Xiaoyan; Guo, Xingjie; Cui, Yan

    2016-01-01

    D-Amino acids are increasingly being recognized as important signaling molecules, and abnormal levels of D-amino acids have been implicated in the pathogenesis of Alzheimer's disease. To evaluate the potential relationship between Alzheimer's disease and D-amino acids, a simple, sensitive, and reliable UPLC-MS/MS method with pre-column derivatization was developed and validated for simultaneous determination of 18 D-amino acids in rat plasma. The analytes were extracted from plasma samples by a protein precipitation procedure, and then derivatized with (S)-N-(4-nitrophenoxycarbonyl) phenylalanine methoxyethyl ester [(S)-NIFE]. Chromatographic separation was achieved using an ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with a mobile phase composed of acetonitrile containing 8 mM ammonium hydrogen carbonate at a flow rate of 0.6 mL min(-1). The analytes were detected by electrospray ionization in positive ion multiple reaction monitoring modes. Under the optimum experimental conditions, all the linear regressions were acquired with r > 0.9932. The limits of quantitation of all derivatized D-amino acids were within 0.05-40.0 ng mL(-1) in rat plasma. The intra- and inter-day precisions, expressed as percentage relative standard deviations (%RSD), were within the range of 12.3 and 10.1%, respectively. The recoveries for all the analytes were observed over the range of 82.8-100.5% with RSD values less than 12.5%. Finally, the proposed method was successfully applied to simultaneous determination of the 18 D-amino acids in plasma from Alzheimer's disease rats and age-matched normal controls. Results showed that the concentrations of D-serine, D-aspartate, D-alanine, D-leucine, and D-proline in Alzheimer's disease rat plasma were significantly decreased compared with those in normal controls, while D-phenylalanine levels increased. It was revealed that some of these D-amino acids would be potential diagnostic biomarkers for Alzheimer's disease.

  14. High throughput LC-MS/MS method for the simultaneous analysis of multiple vitamin D analytes in serum.

    Science.gov (United States)

    Jenkinson, Carl; Taylor, Angela E; Hassan-Smith, Zaki K; Adams, John S; Stewart, Paul M; Hewison, Martin; Keevil, Brian G

    2016-03-01

    Recent studies suggest that vitamin D-deficiency is linked to increased risk of common human health problems. To define vitamin D 'status' most routine analytical methods quantify one particular vitamin D metabolite, 25-hydroxyvitamin D3 (25OHD3). However, vitamin D is characterized by complex metabolic pathways, and simultaneous measurement of multiple vitamin D metabolites may provide a more accurate interpretation of vitamin D status. To address this we developed a high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to analyse multiple vitamin D analytes, with particular emphasis on the separation of epimer metabolites. A supportive liquid-liquid extraction (SLE) and LC-MS/MS method was developed to quantify 10 vitamin D metabolites as well as separation of an interfering 7α-hydroxy-4-cholesten-3-one (7αC4) isobar (precursor of bile acid), and validated by analysis of human serum samples. In a cohort of 116 healthy subjects, circulating concentrations of 25-hydroxyvitamin D3 (25OHD3), 3-epi-25-hydroxyvitamin D3 (3-epi-25OHD3), 24,25-dihydroxyvitamin D3 (24R,25(OH)2D3), 1,25-dihydroxyvitamin D3 (1α,25(OH)2D3), and 25-hydroxyvitamin D2 (25OHD2) were quantifiable using 220μL of serum, with 25OHD3 and 24R,25(OH)2D3 showing significant seasonal variations. This high-throughput LC-MS/MS method provides a novel strategy for assessing the impact of vitamin D on human health and disease.

  15. Validated TLC-densitometry method for the simultaneous analysis of pyrethroid insecticides in agricultural and domestic products

    Directory of Open Access Journals (Sweden)

    Musharraf Syed

    2012-08-01

    Full Text Available Abstract Background Pyrethroids are widely used for the control of pests and insects, as pyrethroids are believed to pose little risk to human health and environment. However, exposure to the pyrethroids exceeding the label directions might adversely affect human health and environment. Hence a careful selection of environment friendly household product is required that must contain exactly the label claimed pyrethroids amount. Results A sensitive and robust TLC-densitometric method for simultaneous quantification of commonly used synthetic pyrethroids including esbiothrin, alpha-cypermethrin and cis/trans permethrin in agricultural and domestic products has been developed and validated. TLC aluminum sheets, precoated with 0.2 mm thick layer of silica gel 60 F-254, were used for chromatographic process. Densitometric analysis of chromatoplates was carried out in absorbance mode at corresponding λmax of each pyrethroid. Equally valid common mobile phase for all pyrethroids consisted of hexane-dichloromethane-ethylacetate-formic acid (8:1.5:0.4:0.1 v/v/v/v which provided sharp and symmetrical peaks of esbiothrin, alpha-cypermethrin, trans-permethrin and cis-permethrin, at Rf 0.31, 0.53, 0.6 and 0.65, respectively. Linear regression data for respective calibration curves showed a good linearity for all pyrethroids with r = 0.991-0.996. Limits of detection (LOD and limits of quantification (LOQ for all pyrethroids were found in the range of 1.6-2.8 and 4.9-8.5 ng/spot, respectively. Conclusions The developed method is applicable for separating the mixture of pyrethroids and at the same time, it is also valid for separating their isomers. The method is reproducible, precise and accurate for the quantitative determination of pyrethroids in agricultural and domestic products.

  16. Deterioration of R-Wave Detection in Pathology and Noise: A Comprehensive Analysis Using Simultaneous Truth and Performance Level Estimation.

    Science.gov (United States)

    Kashif, Muhammad; Jonas, Stephan M; Deserno, Thomas M

    2017-09-01

    For long-term electrocardiography (ECG) recordings, accurate R-wave detection is essential. Several algorithms have been proposed but not yet compared on large, noisy, or pathological data, since manual ground-truth establishment is impossible on such large data. We apply the simultaneous truth and performance level estimation (STAPLE) method to ECG signals comparing nine R-wave detectors: Pan and Tompkins (1985), Chernenko (2007), Arzeno et al. (2008), Manikandan et al. (2012), Lentini et al. (2013), Sartor et al. (2014), Liu et al. (2014), Arteaga-Falconi et al. (2015), and Khamis et al. (2016). Experiments are performed on the MIT-BIH database, TELE database, PTB database, and 24/7 Holter recordings of 60 multimorbid subjects. Existing approaches on R-wave detection perform excellently on healthy subjects (F-measure above 99% for most methods), but performance drops to a range of F = 90.10% (Khamis et al.) to F = 30.10% (Chernenko) when analyzing the 37 million R-waves of multimorbid subjects. STAPLE improves existing approaches (ΔF = 0.04 for the MIT-BIH database and ΔF = 0.95 for the TELE database) and yields a relative (not absolute) scale to compare algorithms' performances. More robust R-wave detection methods or flexible combinations are required to analyze continuous data captured from pathological subjects or that is recorded with dropouts and noise. STAPLE algorithm has been adopted from image to signal analysis to compare algorithms on large, incomplete, and noisy data without manual ground truth. Existing approaches on R-wave detection weakly perform on such data.

  17. Quantitative analysis and simultaneous activity measurements of Cu, Zn-superoxide dismutase in red blood cells by HPLC-ICPMS.

    Science.gov (United States)

    Nuevo Ordoñez, Y; Montes-Bayón, M; Blanco-González, E; Sanz-Medel, A

    2010-03-15

    The interest on accurate and precise determination of metalloproteins such as Cu, Zn-superoxide dismutase (Cu, Zn-SOD) involved in the redox balance of living cells is increasing. For this purpose, analytical strategies that provide absolute protein concentration measurements have to be developed. The determination of Cu, Zn-SOD through the measurement of the Cu associated to the protein, which provides its enzymatic activity, by liquid chromatography with online inductively coupled plasma mass spectrometric (ICPMS) detection is described here. Postcolumn isotope dilution analysis (IDA) of Cu has been applied for quantification after evaluation of the column recovery for the total Cu and also Cu-SOD that turned out to be quantitative. When the concentration results obtained via IDA using high-performance liquid chromatography (HPLC)-ICPMS are plotted versus the activity measurements (using the spectrophotometric pyrogallol autoxidation method) a good correlation curve is obtained. Such results permit us, from ICPMS measurements, to obtain simultaneously the Cu, Zn-SOD absolute concentration as well as its enzymatic activity by interpolation in the previously obtained curve. This possibility was explored in real samples (red blood cells of control individuals and patients with metallic total hip arthroplasty) obtaining a good match between direct enzymatic activity measurements and those obtained by interpolation in the correlation curve. The actual protein identification in the red blood cell extract was conducted by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and two matrixes were compared in order to preserve as much as possible the protein-metal interactions during the MALDI process. Interestingly, using a solution containing trihydroxyacetophenone in citrate buffer permitted us to observe some metal-protein interactions in the MS spectrum of the intact Cu, Zn-SOD from red blood cells.

  18. Anatomic and histological analysis in a goat model used for maxillary sinus floor augmentation with simultaneous implant placement.

    Science.gov (United States)

    Derong, Zou; Lian, Guo; Jiayu, Lu; Xiuli, Zhang; Zhiyuan, Zhang; Xinquan, Jiang

    2010-01-01

    The aim of the present study was to carry out an anatomic survey on the goat maxillary sinus in order to provide accurate and definite anatomic parameters for the design of sinus floor elevation and dental implantation studies in this valuable preclinic animal model. The anatomic topographic structure of the maxillary sinuses was studied bilaterally in 10 adult goats by a gross survey as well as a histological analysis with parasagittal or coronal sections. Then following parameters were defined and measured: (1) maxillary alveolar height (MAH): vertical height from the alveolar crest to the sinus floor; (2) sinus lateral floor width (SLFW): horizontal distance from the lateral border of the anteroposterior bone crest to the sinus lateral wall; (3) infraorbital canal diameter (ICD); and (4) maxillary sinus volume (MSV): the volume occupied by water injected into the sinus. The data were presented with mean + or - SD on both sides. The goat has a maxillary sinus similar to humans, with a slender pyramidal shape that pneumatizes the entire maxilla, and a sinus wall covered with a mucosal lining. From the maxillary sinus floor, there is an anteroposterior bone crest protruding with the infraorbital canal enveloped. It divides the maxillary sinus floor into two parts. The SLFW of the lateral part of the maxillary sinus floor becomes broader, about 5.905 + or - 1.475 mm in the third premolar site, and the MAH increases towards the posterior area, where the maxillary sinus floor is close to the related teeth roots. According to original metrical data, we also proposed a possible operation procedure for sinus floor augmentation. There is enough space in the lateral floor of the maxillary sinus for dental implantation, and the third premolar area might be a suitable position suggested for maxillary sinus augmentation with simultaneous implant placement in a goat model.

  19. Practical approaches to principal component analysis for simultaneously dealing with missing and censored elements in chemical data.

    Science.gov (United States)

    Stanimirova, I

    2013-09-24

    Multivariate chemical data often contain elements that are missing completely at random and the so-called left-censored elements whose values are only known to be below a definite threshold value (reporting limit). In the last several years, attention has been paid to developing methods for dealing with data containing missing elements and those that can handle data with missing elements and outliers. However, processing data with both missing and left-censored elements is still an ongoing problem. The aim of this work was to investigate which method is most suitable for handling left-censored and missing completely at random elements that are present simultaneously in chemical data by using a comparison of the generalized nonlinear iterative partial least squares (NIPALS(1)) algorithm that has been recently proposed, methods that include uncertainty information like maximum likelihood principal component analysis, MLPCA(2), and replacement methods. The results of the Monte Carlo simulation study for artificial and real data sets showed that substitution with half of the reporting limit can be used when the percentage of left-censored elements per variable is up to 30-40%. The generalized NIPALS algorithm is generally recommended for a large percentage of left-censored elements per variable and particularly when a large number of variables are censored. The expectation-maximization approach applied to data with censored elements substituted with half of the reporting limits can be a strategy for dealing with missing and left-censored elements in data, but if the converge criterion is not fulfilled, then the generalized NIPALS algorithm can be applied. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Wavelet-based analysis of gastric microcirculation in rats with ulcer bleedings

    Science.gov (United States)

    Pavlov, A. N.; Rodionov, M. A.; Pavlova, O. N.; Semyachkina-Glushkovskaya, O. V.; Berdnikova, V. A.; Kuznetsova, Ya. V.; Semyachkin-Glushkovskij, I. A.

    2012-03-01

    Studying of nitric oxide (NO) dependent mechanisms of regulation of microcirculation in a stomach can provide important diagnostic markers of the development of stress-induced ulcer bleedings. In this work we use a multiscale analysis based on the discrete wavelet-transform to characterize a latent stage of illness formation in rats. A higher sensitivity of stomach vessels to the NO-level in ill rats is discussed.

  1. Proteomics analysis of cerebral cortex in Wistar rats

    Institute of Scientific and Technical Information of China (English)

    Xiaofeng ZHAO; Jingrong WEN; Shu WANG; Xuemin SHI

    2008-01-01

    To analyze the protein expression pattern of the cerebral cortex in Wistar rats using the proteomics approach, proteins were separated by two-dimensional gel electrophoresis, stained with Coomassie brilliant blue and digested with trypsin. Then, we analyzed the peptide section using a matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and identified the protein by indexing special database (SwissProt) according to the finger printing of the peptide quality. Eighty-four protein spots were identified, includ-ing metabolic enzymes, skeleton proteins, heat shock pro-teins, antioxidant proteins, signaling proteins, proteasome related proteins, neuron and glial specific proteins and serum associated proteins. The result of this study enriches the database of the proteome in the cerebral cortex of rats and lays a foundation for further research of neurological disorders in rat models.

  2. Simultaneous analysis of carbohydrates and organic acids by HPLC-DAD-RI for monitoring goat's milk yogurts fermentation.

    Science.gov (United States)

    da Costa, Marion Pereira; Frasao, Beatriz da Silva; Lima, Bruno Reis Carneiro da Costa; Rodrigues, Bruna Leal; Conte Junior, Carlos Adam

    2016-05-15

    During yogurt manufacture, the lactose fermentation and organic acid production can be used to monitor the fermentation process by starter cultures and probiotic bacteria. In the present work, a simple, sensitive and reproducible high-performance liquid chromatography with dual detectors, diode array detector and refractive index was validated by simultaneous analysis of carbohydrates and organic acids in goat milk yogurts. In addition, pH and bacterial analysis were performed. Separation of all the compounds was performed on an Aminex HPX-87H column (300×7.8 mm, 9 µm) utilizing a 3 mmol L(-1) sulfuric acid aqueous mobile phase under isocratic conditions. Lactose, glucose, galactose, citric, lactic and formic acids were used to evaluate the following performance parameters: selectivity, linearity, precision, limit of detection (LOD), limit of quantification (LOQ), decision limits (CCα), detection capabilities (CCβ), recovery and robustness. For the method application a six goat milk yogurts were elaborated: natural, probiotic, prebiotic, symbiotic, cupuassu fruit pulp, and probiotic with cupuassu fruit pulp. The validated method presented an excellent selectivity with no significant matrix effect, and a broad linear study range with coefficients of determination higher than 0.995. The relative standard deviation was lower than 10% under repeatability and within-laboratory reproducibility conditions for the studied analytes. The LOD of the method was defined from 0.001 to 0.003 µg g(-1), and the LOQ from 0.003 to 0.013 µg g(-1). The CCα was ranged from 0.032 to 0.943 µg g(-1), and the CCβ from 0.053 to 1.604 µg g(-1). The obtained recovery values were from 78% to 119%. In addition, the method exhibited an appropriate robustness for all parameter evaluated. Base in our data, it was concluded that the performance parameters demonstrated total method adequacy for the detection and quantification of carbohydrates and organic acids in goat milk yogurts. The

  3. Correlative Analysis of Behavioral and Physiological Concomitants of Labor in Pregnant Rats

    Science.gov (United States)

    Baer, L. A.; Wade, C. E.; Ronca, A. E.; Dalton, Bonnie (Technical Monitor)

    2002-01-01

    During parturition, rats exhibit characteristic behavioral expressions of labor. Lordosis contractions, consisting of an elongation of the dams body, are observed beginning several hours prior to neonate births, whereas vertical contractions, repeated rapid lifts of the abdomen, occur immediately preceding the birth of each neonate. We analyzed underlying changes in intrauterine pressure (IUP) using a telemetric sensor that we modified for use in freely-moving rats. This technique enabled us to correlate behavioral expressions of labor contractions with IUP. A small telemetric blood pressure sensor was fitted within a fluid-filled balloon, similar in size to a full term rat fetus. On Gestational day 19 of the rats' 22-day pregnancy, a unit was surgically implanted within the uterus. The dams were simultaneously videotaped, enabling us to directly correlate IUP signals with behavioral expressions of labor contractions. Earlier phases of labor, consisting predominantly of lordosis contractions were characterized by lower pressures relative to later phases during which higher pressures and vertical contractions were frequently observed.

  4. Effect of Major Royal Jelly Proteins on Spatial Memory in Aged Rats: Metabolomics Analysis in Urine.

    Science.gov (United States)

    Chen, Di; Liu, Fang; Wan, Jian-Bo; Lai, Chao-Qiang; Shen, Li-Rong

    2017-04-19

    Royal jelly (RJ) produced by worker honeybees is the sole food for the queen bee throughout her life as well as the larvae of worker bees for the first 3 days after hatching. Supplementation of RJ in the diet has been shown to increase spatial memory in rodents. However, the key constituents in RJ responsible for improvement of cognitive function are unknown. Our objective was to determine if the major royal jelly proteins (MRJPs) extracted from RJ can improve the spatial memory of aged rats. The spatial memory assay using the Morris water maze test was administered once to rats after a 14-week feeding. Metabolomics analysis based on quadrupole time-of-flight mass spectrometry was conducted to examine the differences in compounds from urine. Aged male rats fed MRJPs showed improved spatial memory up to 48.5% when compared to the control male aged rats fed distilled water. The metabolite pattern of the MRJPs-fed aged rats was regressed to that of the young rats. Compounds altered by MRJPs were mapped to nicotinate and nicotinamide metabolism, cysteine taurine metabolism, and energy metabolism pathways. In summary, MRJPs may improve spatial memory and possess the potential for prevention of cognitive impairment via the cysteine and taurine metabolism and energy metabolism pathways in aged rats.

  5. Method validation and simultaneous determination of retinol, retinyl palmitate, β-carotene, α-tocopherol and vitamin C in rat serum treated with 7,12 dimethylbenz[a]anthracene and Plantago major L. by high- performance liquid chromatography using diode-array detection.

    Science.gov (United States)

    Levent, Abdulkadi; Oto, Gokhan; Ekin, Suat; Berber, Ismet

    2013-02-01

    A new and simple high-performance liquid chromatography method was developed and validated for the simultaneous determination of retinol, retinyl palmitate, β-carotene, α-tocopherol and vitamin C in rat serum treated with Plantago Major L. and 7,12 dimethylbenz[a]anthracene. High-performance liquid chromatography analysis was performed utilizing an Inertsil ODS3 reversed phase column with methanol-tetrahydrofuran-water as mobile phase under gradient conditions, at 1.5 mL min(-1) flow rate and 25 °C. Diode-array detection was at 325, 450, 290 and 270 nm (retinol and retinyl palmitate), β-carotene, α-tocopherol and vitamin C, respectively and runnig time 18 min. The high-performance liquid chromatography assay and extraction procedure proposed are simple, rapid, sensitive and accurate. The method was then applied for the determination of retinol, retinyl palmitate, β-carotene, α-tocopherol and vitamin C in rat serum. Results of this study demonstrated that; at 60th day DMBA-treated group, there was a significant decrease in vitamin levels compared to the levels of control group. A significant increase was observed in vitamin levels of 7,12 dimethylbenz[α]anthracene+Plantago Major L.-treated group compared to the DMBA-treated group. Additionally, the results obtained in the study are found to be in agreement with data reported in the literature.

  6. Proteomic Analysis of Rat Hippocampus under Simulated Microgravity

    Science.gov (United States)

    Wang, Yun; Li, Yujuan; Zhang, Yongqian; Liu, Yahui; Deng, Yulin

    It has been found that microgravity may lead to impairments in cognitive functions performed by CNS. However, the exact mechanism of effects of microgravity on the learning and memory function in animal nervous system is not elucidated yet. Brain function is mainly mediated by membrane proteins and their dysfunction causes degeneration of the learning and memory. To induce simulated microgravity, the rat tail suspension model was established. Comparative O (18) labeling quantitative proteomic strategy was applied to detect the differentially expressed proteins in rat brain hippocampus. The proteins in membrane fraction from rat hippocampus were digested by trypsin and then the peptides were separated by off-gel for the first dimension with 24 wells device encompassing the pH range of 3 - 10. An off-gel fraction was subjected into LC-ESI-QTOF in triplicate. Preliminary results showed that nearly 77% of the peptides identified were specific to one fraction. 676 proteins were identified among which 108 proteins were found differentially expressed under simulated microgravity. Using the KOBAS server, many enriched pathways, such as metabolic pathway, synaptic vesicle cycle, endocytosis, calcium signaling pathway, and SNAREs pathway were identified. Furthermore, it has been found that neurotransmitter released by Ca (2+) -triggered synaptic vesicles fusion may play key role in neural function. Rab 3A might inhibit the membrane fusion and neurotransmitter release. The protein alteration of the synaptic vesicle cycle may further explain the effects of microgravity on learning and memory function in rats. Key words: Microgravity; proteomics; synaptic vesicle; O (18) ({}) -labeling

  7. A sensitive liquid chromatographic-mass spectrometric method for simultaneous quantification of six iridoid glycosides from Zhi-zi-chi Decoction in rat plasma and its application to a pharmacokinetic study.

    Science.gov (United States)

    Qu, Kankan; Dai, Jinna; Zhao, Longshan; Lu, Yanan; Li, Bin; Zhao, Xu; Hou, Pengyi; Zhang, Yuanting; Bi, Kaishun; Chen, Xiaohui

    2013-05-01

    A sensitive liquid chromatographic-mass spectrometric (LC-MS) method was developed and validated for simultaneous determination of geniposide, geniposidic acid, scandoside methyl ester, gardenoside, deacetyl asperulosidic acid methyl ester and genipin-1-β-gentiobioside after oral administration of Zhi-zi-chi Decoction in rat plasma. The six iridoid glycosides were extracted from plasma samples by protein precipitation, and then separated on an Apollo C18 column (250 mm × 4.6mm, 5 μm) through the application of a gradient elution. The analytes were monitored in positive electrospray ionization by selected ion monitoring mode (SIM). The lower limits of quantitation (LLOQ) of the six analytes were all lower than 6 ng/mL. The accuracy (relative error, RE%) was between -7.0% and 9.9%, while the intra- and inter-day precisions (relative standard deviation, RSD%) were less than 6.3% and 9.8% for the six analytes, respectively. The developed method was successfully applied to a comparative pharmacokinetic study of the six iridoids in rat plasma after oral administration of Zhi-zi-chi Decoction and Gardenia jasminoides extract.

  8. Simultaneous determination of corosolic acid and euscaphic acid in the plasma of normal and diabetic rat after oral administration of extract of Potentilla discolor Bunge by high-performance liquid chromatography/electrospray ionization mass spectrometry.

    Science.gov (United States)

    Li, Jing-jing; Li, Yi; Bai, Min; Tan, Jing-fu; Wang, Qiang; Yang, Jie

    2014-05-01

    Potentilla discolor Bunge has been used for diabetes in China for a long time. Corosolic acid (CA) and euscaphic acid (EA), with significant anti-diabetic activity, are two major triterpenoids in P. discolor. In this study, a specific, sensitive and convenient LC-MS method has been developed for simultaneous determination of CA and EA in the plasma of normal and diabetic rats after oral administration of the extract of P. discolor. The chromatographic separation was achieved using an Alltima C18 column (53 × 7.0 mm, i.d., 3 µm) with a mobile phase composed of 0.1% formic acid water and 0.1% formic acid acetonitrile at a flow rate of 1.0 mL/min. The detection was performed by MS with electrospray ionization interface in negative selected ion monitoring mode. All the validation data, such as specificity, linearity (r(2)  > 0.9991 within 0.025-10.0 µg/mL), lower limit of quantitation (2.5 ng/mL), precision (intra- and inter-day <14.7%), accuracy (<15.0%), recovery (85.7-110.8%) and stability were determined and all of them were within the required limits. This method was successfully applied for the evaluation of the pharmacokinetic behaviors of these two compounds in the plasma of normal and diabetic rats.

  9. A sensitive and selective liquid chromatography-tandem mass spectrometry method for simultaneous determination of five isoquinoline alkaloids from Chelidonium majus L. in rat plasma and its application to a pharmacokinetic study.

    Science.gov (United States)

    Zhou, Qiuhong; Liu, Youping; Wang, Xin; Di, Xin

    2013-01-01

    Chelidonium majus L. is one of the most important medicinal plants of the family Papaveraceae. Its pharmacological effects have been primarily attributed to the presence of a number of alkaloids. In the present study, a sensitive and selective liquid chromatography-tandem mass spectrometry method for simultaneous determination of five isoquinoline alkaloids from Chelidonium majus L. was developed and validated. The analytes (protopine, chelidonine, coptisine, sanguinarine and chelerythrine), together with the internal standard (palmatine), were extracted from acidified rat plasma with ethyl acetate-dichloromethane (4:1, v/v). Chromatographic separation was carried out on a Diamonsil C(18) column with an isocratic mobile phase consisting of acetonitrile and water (adjusted to pH 2.3 with formic acid) (30:70, v/v) at a flow rate of 0.4 ml/min. Mass spectrometric detection was performed by selected reaction monitoring mode via electrospray ionization source operating in positive ionization mode. The assay exhibited good linearity (r ≥ 0.9933) for all the analytes. The lower limits of quantification were 0.197-1.27 ng/ml using only 50 µl of plasma sample. The intra- and inter-day precisions were less than 11.9%, and the accuracy was between -6.3% and 9.3%. The method was successfully applied to the pharmacokinetic study of the five alkaloids in rats after intragastric administration of Chelidonium majus L. extract. Copyright © 2013 John Wiley & Sons, Ltd.

  10. Ultra-fast LC-ESI-MS/MS method for the simultaneous determination of six highly toxic Aconitum alkaloids from Aconiti kusnezoffii radix in rat plasma and its application to a pharmacokinetic study.

    Science.gov (United States)

    Liu, Jingjing; Li, Qing; Yin, Yidi; Liu, Ran; Xu, Huarong; Bi, Kaishun

    2014-01-01

    A fast, sensitive, and efficient ultra-fast LC-ESI-MS/MS method was developed for the simultaneous quantitation of six highly toxic Aconitum alkaloids, that is, aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine, and benzoylhypaconine, in rat plasma after oral administration of crude ethanol extracts from Aconiti kusnezoffii radix by ultrasonic extraction, reflux extraction for 1 h, and reflux extraction for 3 h, respectively. The separation of six Aconitum alkaloids and aminopyrine (internal standard) was performed on an InertSustain® C18 column, and the quantification of the analytes was performed on a 4000Q ultra-fast LC-MS/MS system with turbo ion spray source in the positive ion and multiple-reaction monitoring mode. Absolute recoveries ranged within 65.06-85.1% for plasma samples. The intra- and interday precision and accuracy of analytes were satisfactory. The methods were validated with sensitivity reaching the lower LOQ for aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine, and benzoylhypaconine, which were 0.025, 0.025, 0.050, 0.025, 0.025, and 0.100 ng/mL, respectively. The method was successfully applied to a pharmacokinetic study of six Aconitum alkaloids in rat plasma after oral administration of crude ethanol extracts from the raw root of Aconitum kusnezoffii Reichb. by three different extraction processes.

  11. Simultaneous determination of calycosin-7-O-β-d-glucoside, calycosin, formononetin, astragaloside IV and schisandrin in rat plasma by LC-MS/MS: application to a pharmacokinetic study after oral administration of Shenqi Wuwei chewable tablets.

    Science.gov (United States)

    Sun, Xuehui; Zhang, Pingping; Wu, Xiujun; Wu, Qiong; Zhang, Mengmeng; An, Ye; Shi, Guobing

    2014-08-01

    A rapid, sensitive and reliable high-performance liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous quantification of the five main bioactive components, calycosin, calycosin-7-O-β-d-glucoside, formononetin, astragaloside IV and schisandrin in rat plasma after oral administration of Shenqi Wuwei chewable tablets. Plasma samples were extracted using solid-phase extraction separated on a CEC18 column and detected by MS with an electrospray ionization interface in multiple-reaction monitoring mode. Calibration curves offered linear ranges of two orders of magnitude with r > 0.995. The method had a lower limit of quantitation of 0.1, 0.02, 0.1, 1 and 0.1 ng/mL for calycosin, calycosin-7-O-β-d-glucoside, formononetin, astragaloside IV and schisandrin, respectively. Intra- and inter-day precisions (relative standard deviation) for all analytes ranged from 0.97 to 7.63% and from 3.45 to 10.89%, respectively. This method was successfully applied to the pharmacokinetic study of the five compounds in rats after oral administration of Shenqi Wuwei chewable tablets.

  12. Simultaneous determination of 14-thienyl methylene matrine and matrine in rat plasma by high-performance liquid chromatography-tandem mass spectrometry and its application in a pharmacokinetic study.

    Science.gov (United States)

    Jiang, Minjie; Wang, Lisheng; Jiang, Weizhe; Huang, Shulin

    2015-01-01

    A rapid, sensitive and selective high-performance liquid chromatography-tandem mass spectrometric method (HPLC-MS) has been developed and validated for the simultaneous determination of 14-thienyl methylene matrine (TMM) and matrine (MT) in rat plasma in the present study. The analytes were separated on a C18 column (1.9 μm, 2.1 mm × 100 mm) with a security guard C18 column (5 μm, 2.1 mm × 10 mm) and a triple-quadrupole mass spectrometry equipped with an electrospray ionization (ESI) source was applied for detection. With pseudoephedrine hydrochloride as internal standard, sample pretreatment involved in a one-step protein precipitation with isopropanol:ethyl acetate (v/v, 20:80). The method was linear over the concentration ranges of 5-1000 ng/ml for TMM and 10-2000 ng/ml for MT. The intra-day and inter-day relative standard deviations (RSD) were less than 15% and the relative errors (RE) were all within 15%. The proposed method enables unambiguous identification and quantification of TMM and MT in vivo. This was the first report on determination of the TMM and MT in rat plasma after oral administration of TMM. The results provided a meaningful basis for evaluating the clinical applications of the medicine.

  13. Development of a UFLC-MS/MS method for the simultaneous determination of seven tea catechins in rat plasma and its application to a pharmacokinetic study after administration of green tea extract.

    Science.gov (United States)

    Huo, Yanshuang; Zhang, Qian; Li, Qing; Geng, Bingjie; Bi, Kaishun

    2016-06-05

    A rapid, sensitive and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed and validated for simultaneous determination of seven green tea catechins including catechin (C), (-)-epicatechin (EC), gallocatechin (GC), (-)-epigallocatechin (EGC), (-)-epicatechin-3-gallate (ECG), gallocatechin-3-gallate (GCG) and (-)-epigallocatechin-3-gallate (EGCG) in rat plasma. The plasma samples were firstly hydrolysed with the mixture of β-glucuronidase and sulfatase, which were then extracted by liquid-liquid extraction with ethyl acetate-isopropanol (1:1, v/v). The analytes were separated on a Venusil MP C18 column (Venusil, China) with a gradient elution at a flow rate of 0.4mL/min. The detection was performed in negative ionization and multiple reaction monitoring (MRM) mode. All the calibration curves exhibited good linearity (r>0.9943) with intra- and inter-day precisions of less than 14.3% and the accuracy deviations ranging from -8.8% to 7.5%. The extraction recoveries of the analytes and ethyl gallate (internal standard) were all more than 72%. The validated method was successfully applied to a pharmacokinetic study of seven catechins in rat plasma after oral administration of the green tea extract at different doses of 0.4, 1.2 and 2.0g/kg.

  14. Simultaneous determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and geniposide in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study after administration of Reduning injection.

    Science.gov (United States)

    Wang, Yanjuan; Wen, Jing; Zheng, Weihua; Zhao, Longshan; Fu, Xiaohuan; Wang, Zhenzhong; Xiong, Zhili; Li, Famei; Xiao, Wei

    2015-01-01

    A simple, specific and sensitive ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established and validated for simultaneous determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and geniposide in rat plasma using puerarin as an internal standard (IS). Plasma samples were pretreated by a one-step direct protein precipitation procedure with acetonitrile after acidified using as little as 50 μL plasma. Chromatographic separation was performed on an Acquity BEH C18 column (100 × 2.1 mm, 1.7 µm) at a flow rate of 0.2 mL/min by a gradient elution, using 0.2% acetic acid-methanol as mobile phase. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring via electrospray ionization source with negative ion mode. Calibration curves showed good linearity (r > 0.995) over wide concentration ranges. The intra- and inter-day precisions were <15%, and the accuracy was within ±8.0%. The validated method was successfully applied to a pharmacokinetic study of the four bioactive components in rats after intravenous administration of Reduning injection.

  15. Simultaneous determination of six alkaloid components in rat plasma and its application to pharmacokinetic study of Danmu preparations by an ultra fast liquid chromatography-electrospray ionization-tandem mass spectrometry.

    Science.gov (United States)

    Yin, Rong; Chen, Jiaquan; Zhao, Yonggang; Jia, Xiaobin; Zhang, Zhiyuan; Feng, Liang; Wang, Hui; Wang, Jingjing; Zhu, Fenxia

    2015-03-01

    Danmu injection and Danmu tablet are two widely used traditional Chinese medicine made of Nauclea officinalis (commonly known as Danmu), in which the alkaloids are the major active substances. In this paper, an ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed for simultaneous determination and the pharmacokinetic characteristics study of six main active alkaloids (naucleamide A-10-O-β-d-glucopyranosid, naucleamide G, pumiloside, 3-epi-pumiloside, strictosamide and vincosamide) of the two above-mentioned Danmu preparations in rat plasma. In the course of the experiment, following sample preparation by protein precipitation with methanol-ethyl acetate (2:1, v/v), the nitrogen-dried extraction was reconstituted in methanol and assayed on a C18 column using a gradient elution program with mobile phase consisting of acetonitrile and water containing 0.1% formic acid. The MS detection was performed in positive ionization mode with selected ion transitions. The established method was fully validated and proved to be sensitive and specific with lower limits of quantification (LLOQs) all less than 0.32ng/mL in rat plasma and matrix effects ranged from 88.87 to 108.27%. Good linearities of six alkaloids were obtained in respective concentration ranges (r(2)>0.995). The average extract recoveries for each compound at three quality control concentration levels were no less than 79.70%, and the precision and accuracy were within the acceptable limits. The validated method was successfully applied to the pharmacokinetic study of six alkaloid components of Danmu injection and tablet in rat plasma. The obtained results may be helpful to reveal the action mechanism and guide the clinical application of Danmu preparations.

  16. Simultaneous and in situ analysis of thermal and volumetric properties of starch gelatinization over wide pressure and temperature ranges.

    Science.gov (United States)

    Randzio, Stanislaw L; Orlowska, Marta

    2005-01-01

    A method for simultaneous and in situ analysis of thermal and volumetric properties of starch gelatinization from 0.1 to 100 MPa and from 283 to 430 K is described. The temperature of a very sensitive calorimetric detector containing a starch-water emulsion at a selected pressure is programmed to rise at a slow rate; volume variations are performed automatically to keep the selected pressure constant while the heat exchange rate and the volume are recorded. The method is demonstrated with a novel investigation of pressure effects on a sequence of three phase transitions in an aqueous emulsion of wheat starch (56 wt % water). The volume changes during the main endothermic transition (M), associated with melting of the crystalline part of the starch granules and a helix-coil transformation in amylopectin, but also with an important swelling, were separated into a volume increase associated with swelling and a volume decrease associated with the transition itself. Thermodynamic parameters for this transition together with their pressure dependencies have been obtained from four independent experiments at each pressure. The data are thermodynamically consistent, but are poorly described by the Clapeyron equation. The negative volume change of the slow exothermic transition (A) appearing just after the main endothermic transition (M) is small, spread out over a wide temperature interval, and occurs at higher temperatures with increasing pressures. This transition is probably associated with reassociation of the unwound helixes of amylopectin with parts of amylopectin molecules other than their original helix duplex partner. The positive volume change of the high-temperature, endothermic transition (N) with a small enthalpy change is probably associated with a nematic-isotropic transformation ending the formation of a homogeneous SOL phase (in the sense of Flory), and is also pushed to higher temperatures with increasing pressures. Knowledge of the state of wheat starch

  17. A sensitive LC-MS/MS method for simultaneous determination of six flavonoids in rat plasma: application to a pharmacokinetic study of total flavonoids from mulberry leaves.

    Science.gov (United States)

    He, Jun; Feng, Ying; Ouyang, Hui-Zi; Yu, Bin; Chang, Yan-Xu; Pan, Gui-Xiang; Dong, Gai-Ying; Wang, Tao; Gao, Xiu-Mei

    2013-10-01

    A simple and sensitive LC-MS/MS method has been developed and validated for the determination of rutin, isoquercitrin, astragalin, quercetin, kaempferol and isorhamnetin in rat plasma using naringin as the internal standard (IS). The plasma samples were pretreated and extracted by liquid-liquid extraction. Chromatographic separation was accomplished on a C18 column with a 10 min gradient elution using acetonitrile and 0.1% formic acid aqueous solution as mobile phase at a flow rate of 0.3 mL min(-1). A tandem mass spectrometric detection was conducted using multiple reaction monitoring (MRM) via an electrospray ionization (ESI) source and operating in the negative ionization mode. The lower limit of quantitation (LLOQ) of each analyte was lower than 1 ng mL(-1). Intra-day and inter-day precisions were less than 11.9%. The relative errors of accuracy were in the range of -9.2% to 6.1%. The mean recoveries of flavonoids and IS were higher than 53.8%. The proposed method was further applied to investigate the pharmacokinetics of all analytes after a single oral administration of total flavonoids from mulberry leaves to rats.

  18. Sensitive liquid chromatography/tandem mass spectrometry method for the simultaneous determination of olanzapine, risperidone, 9-hydroxyrisperidone, clozapine, haloperidol and ziprasidone in rat brain tissue.

    Science.gov (United States)

    Zhang, Guodong; Terry, Alvin V; Bartlett, Michael G

    2007-10-15

    One prerequisite for therapeutic effects of psychiatric drugs is the ability to pass the blood brain barrier. Hence, it is important to know the concentration of antipsychotic drugs in brain tissue. In general, determinations of lipophilic compounds from lipophilic matricies such as the brain are a challenge. Here we have adapted a plasma assay for antipsychotics for the target organ the brain. Using modified sample preparation and chromatographic strategies, the analytes were extracted from rat brain homogenate and analyzed by LC-MS/MS. The method used a Waters Atlantis dC-18 (30 mm x 2.1 mm i.d., 3 microm) column with a mobile phase of acetonitrile/5 mM ammonium formate (pH 6.1 adjusted with formic acid) and gradient elution. All analytes were detected in positive ion mode using multiple-reaction monitoring. The method was validated and the linearity, lower limit of quantitation, precision, accuracy, recoveries, specificity and stability were determined. This method was then successfully used to quantify the rat brain tissue concentration of the analytes after chronic treatment with these antipsychotic drugs.

  19. Simultaneous determination of a novel diphenylpiperazine calcium channel blocker and its four metabolites in rat liver microsomes by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Guo, Wei; Kong, Dezhi; Du, Yingfeng; Shi, Xiaowei; Wang, Wei; Wang, Yongli

    2012-01-01

    Dipfluzine hydrochloride (Dip), a novel diphenylpiperazine calcium channel blocker, has revealed the characteristics of a promising candidate for the treatment of cerebral vascular diseases in preclinical studies. Our research identified and quantified Dip and its 4 metabolites (M1, M2, M4 and M5) in rat liver microsomes by liquid chromatography tandem mass spectrometry. The results showed that Dip was firstly metabolized to M1 and M5 by 1- and 4-dealkylation from a piperazine nitrogen, and then the latter was subsequently metabolized to M2 and M4. The concentrations of Dip, M1, M2 and M5 were 557.3 ± 26.3, 854.3 ± 46.0, 2796.7± 126.9, 2473.3 ± 82.6 and 4.0 ± 0.4, 2.4 ± 0.1, 318.2 ± 8.7 and 27.4 ± 1.5 ng/ml in male and female rats, respectively. M4 (404.2 ± 22.2 ng/ml) was detected only in males not in females, suggesting that there is gender difference in the metabolism of Dip.

  20. A sensitive and specific liquid chromatography mass spectrometry method for simultaneous determination of berberine, palmatine, coptisine, epiberberine and jatrorrhizine from Coptidis Rhizoma in rat plasma

    Science.gov (United States)

    Yu, Sen; Pang, Xiaoyan; Deng, Yuanxiong; Liu, Li; Liang, Yan; Liu, Xiaodong; Xie, Lin; Wang, Guangji; Wang, Xinting

    2007-11-01

    A sensitive and specific liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method has been developed and validated for the identification and quantification of five protoberberine alkaloids, which are berberine, palmatine, coptisine, epiberberine and jatrorrhizine, in rat plasma using tetrahydroberberine as an internal standard. Following solid-phase extraction, the analytes were separated by linear gradient elution on a Shim-pack ODS (4.6 [mu]m, 150 mm × 2.0 mm i.d.) column and analyzed in selected ion monitoring (SIM) mode with a positive electrospray ionization (ESI) interface using the respective [M]+ and [M + H]+ ions, [M]+ = 336 for berberine; 320 for coptisine; 336 for epiberberine; 338 for jatrorrhizine; 352 for palmatine and [M + H]+ = 340 for the internal standard. The method was validated over the concentration range of 0.31-20 ng mL-1 for all the five protoberberine alkaloids. Within-batch and between-batch precisions (R.S.D.%) were all within 15% and accuracy (%Er) ranged from -5 to 5%. The lower limits of quantification were 0.31 ng mL-1 for all analytes. The extraction recoveries were on average 80.8% for berberine, 67E0% for coptisine, 66.2% for epiberberine, 71.8% for jatrorrhizine and 73E2% for palmatine. The validated method was used to study the pharmacokinetic profile of the five protoberberine alkaloids in rat plasma after oral administration of Coptidis Rhizoma extract.

  1. Multivariate analysis of DSC-XRD simultaneous measurement data: a study of multistage crystalline structure changes in a linear poly(ethylene imine) thin film.

    Science.gov (United States)

    Kakuda, Hiroyuki; Okada, Tetsuo; Otsuka, Makoto; Katsumoto, Yukiteru; Hasegawa, Takeshi

    2009-01-01

    A multivariate analytical technique has been applied to the analysis of simultaneous measurement data from differential scanning calorimetry (DSC) and X-ray diffraction (XRD) in order to study thermal changes in crystalline structure of a linear poly(ethylene imine) (LPEI) film. A large number of XRD patterns generated from the simultaneous measurements were subjected to an augmented alternative least-squares (ALS) regression analysis, and the XRD patterns were readily decomposed into chemically independent XRD patterns and their thermal profiles were also obtained at the same time. The decomposed XRD patterns and the profiles were useful in discussing the minute peaks in the DSC. The analytical results revealed the following changes of polymorphisms in detail: An LPEI film prepared by casting an aqueous solution was composed of sesquihydrate and hemihydrate crystals. The sesquihydrate one was lost at an early stage of heating, and the film changed into an amorphous state. Once the sesquihydrate was lost by heating, it was not recovered even when it was cooled back to room temperature. When the sample was heated again, structural changes were found between the hemihydrate and the amorphous components. In this manner, the simultaneous DSC-XRD measurements combined with ALS analysis proved to be powerful for obtaining a better understanding of the thermally induced changes of the crystalline structure in a polymer film.

  2. Qualitative and quantitative analysis of bones in adult rats by repeated intraperitoneal administration of paclitaxel (taxol) using radioisotope EDXRF technique

    Energy Technology Data Exchange (ETDEWEB)

    Oezdemir, Y. E-mail: yozdemir25@yahoo.com; Iyiguen, Ibrahim; Durak, Ridvan

    2003-05-15

    Energy dispersive X-ray fluorescence analysis of right back leg bone samples taken from adult female rats administered paclitaxel (taxol) was carried out using standard addition method. Because qualitative and quantitative data analysis of rat bone samples is not found in the literature, comparison was not made with other experimental results. Our experimental results are presented and discussed in this study.

  3. Gene expression profile analysis of genes in rat hippocampus from antidepressant treated rats using DNA microarray

    Directory of Open Access Journals (Sweden)

    Shin Minkyu

    2010-11-01

    Full Text Available Abstract Background The molecular and biological mechanisms by which many antidepressants function are based on the monoamine depletion hypothesis. However, the entire cascade of mechanisms responsible for the therapeutic effect of antidepressants has not yet been elucidated. Results We used a genome-wide microarray system containing 30,000 clones to evaluate total RNA that had been isolated from the brains of treated rats to identify the genes involved in the therapeutic mechanisms of various antidepressants, a tricyclic antidepressant (imipramine. a selective serotonin reuptake inhibitor (fluoxetine, a monoamine oxidase inhibitor (phenelzine and psychoactive herbal extracts of Nelumbinis Semen (NS. To confirm the differential expression of the identified genes, we analyzed the amount of mRNA that was isolated from the hippocampus of rats that had been treated with antidepressants by real-time RT-PCR using primers specific for selected genes of interest. These data demonstrate that antidepressants interfere with the expression of a large array of genes involved in signaling, survival and protein metabolism, suggesting that the therapeutic effect of these antidepressants is very complex. Surprisingly, unlike other antidepressants, we found that the standardized herbal medicine, Nelumbinis Semen, is free of factors that can induce neurodegenerative diseases such as caspase 8, α-synuclein, and amyloid precursor protein. In addition, the production of the inflammatory cytokine, IFNγ, was significantly decreased in rat hippocampus in response to treatment with antidepressants, while the inhibitory cytokine, TGFβ, was significantly enhanced. Conclusions These results suggest that antidepressants function by regulating neurotransmission as well as suppressing immunoreactivity in the central nervous system.

  4. Simultaneous determination of ledipasvir, sofosbuvir and its metabolite in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study.

    Science.gov (United States)

    Pan, Chenwei; Chen, Yongping; Chen, Weilai; Zhou, Guangyao; Jin, Lingxiang; Zheng, Yi; Lin, Wei; Pan, Zhenzhen

    2016-01-01

    In this work, a rapid and sensitive ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the determination of ledipasvir, sofosbuvir and its metabolite GS-331007 in rat plasma was developed. The analytes and the internal standard (diazepam) were separated on an Acquity UPLC BEH C18 chromatography column (2.1mm×50mm, 1.7μm) using gradient elution with a mobile phase of acetonitrile and 0.1% formic acid in water at a flow rate of 0.4mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z 889.8→130.1 for ledipasvir, m/z 530.3→243.1 for sofosbuvir, m/z 261.5→113.1 for GS-331007 and m/z 285.2→193.1 for diazepam (IS) using a positive electrospray ionization interface. The method was validated over a concentration range of 2-500ng/mL for ledipasvir, 10-2000ng/mL for sofosbuvir and 10-2000ng/mL for GS-331007. Total time for each chromatography was 3.0min. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations (RSD)sofosbuvir and GS-331007 in rats.

  5. Simultaneous Determination of Eight Ginsenosides in Rat Plasma by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry: Application to Their Pharmacokinetics

    Directory of Open Access Journals (Sweden)

    Li-Yuan Ma

    2015-12-01

    Full Text Available A high-performance liquid chromatography–electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS method was successfully developed and validated for the identification and determination of eight ginsenosides: ginsenoside Rg1 (1; 20(S-ginsenoside Rh1 (2; 20(S-ginsenoside Rg2 (3; 20(R-ginsenoside Rh1 (4; 20(R-ginsenoside Rg2 (5; ginsenoside Rd (6; 20(S-ginsenoside Rg3 (7; and 20(R-ginsenoside Rg3 (8 in rat plasma. The established rapid method had high linearity, selectivity, sensitivity, accuracy, and precision. The method has been used successfully to study the pharmacokinetics of abovementioned eight ginsenosides for the first time. After an oral administration of total saponins in the stems-leaves of Panax ginseng C. A. Meyer (GTSSL at a dose of 400 mg/kg, the ginsenosides 6, 7, and 8, belonging to protopanaxadiol-type saponins, exhibited relatively long tmax values, suggesting that they were slowly absorbed, while the ginsenosides 1–5, belonging to protopanaxatriol-type saponins, had different tmax values, which should be due to their differences in the substituted groups. Compounds 2 and 4, 3 and 5, 7 and 8 were three pairs of R/S epimerics at C-20, which was interesting that the t1/2 of 20(S-epimers were always longer than those of 20(R-epimers. This pharmacokinetic identification of multiple ginsenosides of GTSSL in rat plasma provides a significant basis for better understanding the clinical application of GTSSL.

  6. Purification and sequence analysis of two rat tissue inhibitors of metalloproteinases

    Science.gov (United States)

    Roswit, W. T.; McCourt, D. W.; Partridge, N. C.; Jeffrey, J. J.

    1992-01-01

    Two protein inhibitors of metalloproteinases (TIMP) were isolated from medium conditioned by the clonal rat osteosarcoma line UMR 106-01. Initial purification of both a 30-kDa inhibitor and a 20-kDa inhibitor was accomplished using heparin-Sepharose chromatography with dextran sulfate elution followed by DEAE-Sepharose and CM-Sepharose chromatography. Purification of the 20-kDa inhibitor to homogeneity was completed with reverse-phase high-performance liquid chromatography. The 20-kDa inhibitor was identified as rat TIMP-2. The 30-kDa inhibitor, although not purified to homogeneity, was identified as rat TIMP-1. Amino terminal amino acid sequence analysis of the 30-kDa inhibitor demonstrated 86% identity to human TIMP-1 for the first 22 amino acids while the sequence of the 20-kDa inhibitor was identical to that of human TIMP-2 for the first 22 residues. Treatment with peptide:N-glycosidase F indicated that the 30-kDa rat inhibitor is glycosylated while the 20-kDa inhibitor is apparently unglycosylated. Inhibition of both rat and human interstitial collagenase by rat TIMP-2 was stoichiometric, with a 1:1 molar ratio required for complete inhibition. Exposure of UMR 106-01 cells to 10(-7) M parathyroid hormone resulted in approximately a 40% increase in total inhibitor production over basal levels.

  7. Purification and sequence analysis of two rat tissue inhibitors of metalloproteinases

    Science.gov (United States)

    Roswit, W. T.; McCourt, D. W.; Partridge, N. C.; Jeffrey, J. J.

    1992-01-01

    Two protein inhibitors of metalloproteinases (TIMP) were isolated from medium conditioned by the clonal rat osteosarcoma line UMR 106-01. Initial purification of both a 30-kDa inhibitor and a 20-kDa inhibitor was accomplished using heparin-Sepharose chromatography with dextran sulfate elution followed by DEAE-Sepharose and CM-Sepharose chromatography. Purification of the 20-kDa inhibitor to homogeneity was completed with reverse-phase high-performance liquid chromatography. The 20-kDa inhibitor was identified as rat TIMP-2. The 30-kDa inhibitor, although not purified to homogeneity, was identified as rat TIMP-1. Amino terminal amino acid sequence analysis of the 30-kDa inhibitor demonstrated 86% identity to human TIMP-1 for the first 22 amino acids while the sequence of the 20-kDa inhibitor was identical to that of human TIMP-2 for the first 22 residues. Treatment with peptide:N-glycosidase F indicated that the 30-kDa rat inhibitor is glycosylated while the 20-kDa inhibitor is apparently unglycosylated. Inhibition of both rat and human interstitial collagenase by rat TIMP-2 was stoichiometric, with a 1:1 molar ratio required for complete inhibition. Exposure of UMR 106-01 cells to 10(-7) M parathyroid hormone resulted in approximately a 40% increase in total inhibitor production over basal levels.

  8. Cadmium toxicity in perinatal rat hepatocytes: Electron microscopy, X-ray microanalysis, and morphometric analysis

    Energy Technology Data Exchange (ETDEWEB)

    Kawahara, A.; Yoshizuka, M.; Hirano, T.; Ohsato, K.; Fujimoto, S. (Univ. of Occupational and Environmental Health, Kitakyushu (Japan))

    1990-10-01

    Effects of cadmium on the fetal and postnatal rat hepatocytes were studied with an electron microscope and an X-ray microanalyzer. Pregnant and lactating Wistar rat dams at 15 and 21 days of pregnancy and at 3 days after delivery received intraperitoneal injections of cadmium sulfate (1 mg/kg body weight) for 3 days. On the day following the last injection, the livers were isolated from the fetal and suckling rats and provided for electron microscopy. The livers from the untreated fetal and newborn rats served as control. Large bile canaliculi, which were formed by five or more hepatocytes, were frequently observed in the cadmium-treated perinatal rat livers. The intercellular space between each adjacent hepatocyte was widened. By X-ray microanalysis, cadmium peaks were preferentially detected out from intramitochondrial granules of the cadmium-treated hepatocytes. By morphometric analysis, the increase both in the mitochondria volume and in the number of intramitochondrial granules was evident in the cadmium-treated hepatocytes when compared to those of control. These data suggest the preferential accumulation of cadmium in mitochondria of the hepatocytes interferes with the morphogenesis of the perinatal rat liver.

  9. Evaluation of non-uniform attenuation correction using simultaneous transmission and emission computed tomography. Basic analysis with myocardial phantom

    Energy Technology Data Exchange (ETDEWEB)

    Otake, Hidenori; Yukihiro, Masashi; Fukushima, Yumi [Gunma Univ., Maebashi (Japan). School of Medicine] [and others

    1996-03-01

    Simultaneous transmission emission protocol (STEP), developed for the non-uniform attenuation correction of single photon emission computed tomography (SPECT) was evaluated using the cardiac phantom prepared with and without a myocardial wall defect. Emission computed tomography (ECT) of the cardiac phantom using {sup 201}Tl was acquired. Transmission data (TCT) were taken using a line source of {sup 99m}Tc. Myocardial images with STEP method were superior in the homogeneity of intramyocardial radioactivity and spatial resolution to the conventional SPECT images. This is an excellent method because of the accurate matching position between TCT and ECT images and shortening the examination time by simultaneous data acquisition. It would be clinically useful for diagnosing various myocardial diseases. (author).

  10. [Evaluation of non-uniform attenuation correction using simultaneous transmission and emission computed tomography--basic analysis with myocardial phantom].

    Science.gov (United States)

    Otake, H; Yukihiro, M; Fukushima, Y; Imai, T; Hosono, K; Hatori, N; Watanabe, N; Hirano, T; Inoue, T; Takahashi, M; Ban, R; Endo, K

    1996-03-01

    Simultaneous transmission emission protocol (STEP), developed for the non-uniform attenuation correction of single photon emission computed tomography (SPECT) was evaluated using the cardiac phantom prepared with and without a myocardial wall defect. Emission computed tomography (ECT) of the cardiac phantom using 201Tl was acquired. Transmission data (TCT) were taken using a line source of 99mTc. Myocardial images with STEP method were superior in the homogeneity of intramyocardial radioactivity and spatial resolution to the conventional SPECT images. This is an excellent method because of the accurate matching position between TCT and ECT images and shortening the examination time by simultaneous data acquisition. It would be clinically useful for diagnosing various myocardial diseases.

  11. Analysis of morphometric and ultrastructural characteristics of blood microcirculation in hippocampus of rats with alloxan diabetes under administration of citicoline

    Directory of Open Access Journals (Sweden)

    Владимир Иванович Жилюк

    2015-07-01

    Full Text Available Aim: determine morphometric and ultrastructural features of blood microcirculation in hippocampus of rats with alloxan diabetes under experimental therapy with citicoline.Materials and methods. The research was carried out on 48 white male Wistar rats (250-300 g randomized in 3 groups by 16 animals: I – intact (distilled water, intragastrically; II – animals with diabetes (distilled water, intragastrically; III – animals with diabetes + citicoline (500 mg/kg, intragastrically. Diabetes was reproduced by single subcutaneous injection of alloxan monohydrate (150 mg/kg. Blood glucose level was determined on the 11th day after administration of alloxan using a glucometer. Citicoline and distilled water were introduced once a day during 20 days starting from the 11th day after administration of alloxan. In morpho-functional study of hippocampal CA1-zone neurons the histological sections were deparaffinized and dyed with gallocyanin-chrome alum by Einarson for specific detection of RNA. Pictures were received with a microscope Axioskop (Zeiss, Germany. Using a computer-based picture analysis system VIDAS-386 (Kontron Elektronik, Germany the density of endothelial nuclei, the nucleus area and the concentration of RNA in the nucleus were determined. The study of ultrastructural changes of blood microcirculation in hippocampus was carried out using transmission electron microscope PEM-100-01 («SELMI», Ukraine by standard procedure and in accordance with common standards. Statistical difference between the groups were assessed by non-parametric Mann-Whitney U-test.Results. It has been established that citicoline contributed to recovery of proliferative activity of endothelium in vascular bed of hippocampus manifested with increase of endothelial nuclei density, their area and levels of nuclear RNA by 19,4% (p < 0,001, 17,2% (p < 0,05 and 25,8% (p < 0,01. Simultaneously it has been determined that hematoencephalic barrier damage in hippocampus of

  12. On-line analysis of middle latency auditory evoked potentials (MLAEP) for monitoring depth of anaesthesia in laboratory rats

    DEFF Research Database (Denmark)

    Jensen, E W; Nygaard, M; Henneberg, S W

    1998-01-01

    using neuromuscular blocking agents (NMBA). A number of studies suggest that the Middle Latency Auditory Evoked Potentials (MLAEP) contain information about the state of consciousness in humans. The purpose of this study was to examine whether the AEP could serve as an indicator of depth of anaesthesia...... and decreasing gradually to a level between 50 and 20 as the rat was anaesthetised. Nine rats were anaesthetised and included in the study. Four doses of Hypnorm vet. and Dormicum were given as a total, each with 5 minutes interval. Clinical signs of the level of anaesthesia were observed simultaneously...... with the AEP. The results showed that in four rats DAI decreased to a level below 30 while anaesthetised. In the remaining five rats the AEP was only decreased to a level below 45. The results indicated that a simple dosing regimen based on weight was unable to give the same depth of anaesthesia in individual...

  13. HYPOLIPEDEMIC EFFECT OF CYNODON DACTYLON ON HISTOPATHOLOGICAL STUDY AND DNA FRAGMENTATION ANALYSIS IN EXPERIMENTALLY INDUCED HYPERCHOLESTEREMIC RATS

    Directory of Open Access Journals (Sweden)

    C. Selva Kumar

    2011-11-01

    Full Text Available Hypercholesteremia is one of the risk factors for coronary artery disease. The present study highlights the efficacy of Ayurvedic herbal formulation Cynodon dactylon (Bermuda grass on histopathological study and DNA fragmentation analysis in experimentally induced hypercholesteremic rats. Four groups of rats were employed namely control, hypercholesterolemia rats (4% Cholesterol+1% cholic acid, Cynodon dactylon treatment in hypercholesteremic rats and Cynodon dactylon alone treated rats. Results of DNA fragmentation was found to be greater in High cholesterol diet fed groups. Lesser fragmentation was found in high cholesterol diet + Cynodon dactylon treated group when compared to the high cholesterol diet group. Cynodon dactylon alone treated group is comparably similar to that of normal group (lesser fragmentation. Histopathological study of thoracic aorta of Cynodon dactylon treated group shows decrease in atherogenicity compared to untreated high cholesterol diet fed rats. The data demonstrated that Cynodon dactylon formulation was associated with hypolipidemic effects on the experimentally induced hypercholesteremic rats.

  14. Simultaneous determination of curcumin diethyl disuccinate and its active metabolite curcumin in rat plasma by LC-MS/MS: Application of esterase inhibitors in the stabilization of an ester-containing prodrug.

    Science.gov (United States)

    Ratnatilaka Na Bhuket, Pahweenvaj; Niwattisaiwong, Nuansri; Limpikirati, Patanachai; Khemawoot, Phisit; Towiwat, Pasarapa; Ongpipattanakul, Boonsri; Rojsitthisak, Pornchai

    2016-10-15

    Four esterase inhibitors, ethylenediamine tetraacetic acid disodium (Na2EDTA), sodium fluoride (NaF), bis(4-nitrophenyl) phosphate (BNPP) and phenylmethanesulfonyl fluoride (PMSF), were evaluated for their inhibitory effects on enzymatic hydrolysis of labile phenolate esters in curcumin diethyl disuccinate (CDD), a prodrug of curcumin (CUR), in rat plasma. BNPP and PMSF at 10mM exhibited stabilization by preventing degradation of CDD. BNPP at a final concentration of 10mM was subsequently selected to prevent ex vivo metabolism of CDD throughout LC-MS/MS analysis of CDD and CUR in rat plasma. A simple protein precipitation technique using acetonitrile as a precipitating agent was used to extract CDD, CUR and dimethylcurcumin (DMC), an internal standard, from rat plasma. Chromatographic separation was performed on a Halo C8 column (4.6×50mm, 2.7μm) using an isocratic mobile phase containing acetonitrile-0.2% formic acid in water (73:27v/v) with a flow rate of 0.4mLmin(-1). An AB SCIEX QTRAP(®) 6500 mass spectrometer was operated using a positive ion electrospray mode for ionization and detection of analytes and internal standard. Calibration curves for CDD and CUR were established using 50μL of rat plasma over the concentration range of 1-500ngmL(-1). The developed method was fully validated according to US Food and Drug Administration (FDA) guidelines for selectivity, sensitivity, linearity, accuracy, precision, dilution integrity, recovery, matrix effect, and stability. The validated method was applied to evaluate the pharmacokinetics of CDD and CUR in rats after a single intravenous dose of 40mgkg(-1). The method using BNPP as an esterase inhibitor was successful in determining the remaining CDD in rat plasma. The pharmacokinetic results indicate that CDD in rats is converted instantaneously to CUR after intravenous administration and a higher CUR plasma concentration at 5min is achieved in comparison with direct intravenous injection of CUR. Copyright © 2016

  15. COMPUTER-ASSISTED SEMEN ANALYSIS OF RAT SPERMATOZOA AFTER AN INTRAPERITONEAL ADMINISTRATION OF INSECTICIDE DIAZINON

    Directory of Open Access Journals (Sweden)

    R. TOMAN

    2013-12-01

    Full Text Available The aim of this study was to reveal the effect of diazinon on the rat spermatozoa motility characteristics using the computer-assisted semen analysis (CASA. Motility, progressive motility, DAP, DCL, DSL, VAP, VCL, VSL, STR, LIN, WOB, ALH, and BCF after the diazinon i.p. administration of 20 mg/kg b.w. were evaluated. 36 hours after the diazinon administration, only slight decrease in VCL, DCL and increase in percentage of progressive motility in the diazinon-treated group. Significant decrease (P<0.01 was only observed in BCF in diazinon-treated group. Computer-assisted semen analysis (CASA of rat sperm motility showed that acute diazinon administration slightly affected the rat sperm motility which can be the first step in the decreased fertilization capacity caused by pesticides. Further investigation of reproductive effects of diazinon is needed.

  16. Multiway analysis methods applied to the fluorescence excitation-emission dataset for the simultaneous quantification of valsartan and amlodipine in tablets

    Science.gov (United States)

    Dinç, Erdal; Ertekin, Zehra Ceren; Büker, Eda

    2017-09-01

    In this study, excitation-emission matrix datasets, which have strong overlapping bands, were processed by using four different chemometric calibration algorithms consisting of parallel factor analysis, Tucker3, three-way partial least squares and unfolded partial least squares for the simultaneous quantitative estimation of valsartan and amlodipine besylate in tablets. In analyses, preliminary separation step was not used before the application of parallel factor analysis Tucker3, three-way partial least squares and unfolded partial least squares approaches for the analysis of the related drug substances in samples. Three-way excitation-emission matrix data array was obtained by concatenating excitation-emission matrices of the calibration set, validation set, and commercial tablet samples. The excitation-emission matrix data array was used to get parallel factor analysis, Tucker3, three-way partial least squares and unfolded partial least squares calibrations and to predict the amounts of valsartan and amlodipine besylate in samples. For all the methods, calibration and prediction of valsartan and amlodipine besylate were performed in the working concentration ranges of 0.25-4.50 μg/mL. The validity and the performance of all the proposed methods were checked by using the validation parameters. From the analysis results, it was concluded that the described two-way and three-way algorithmic methods were very useful for the simultaneous quantitative resolution and routine analysis of the related drug substances in marketed samples.

  17. Biosynthesis and stereoselective analysis of (-)- and (+)-zaltoprofen glucuronide in rat hepatic microsomes and its application to the kinetic analysis.

    Science.gov (United States)

    Wang, Haina; Ji, Jianbo; Zeng, Su

    2011-08-15

    Zaltoprofen, available commercially as a racemic mixture, is a propionic acid derivative of non-steroidal anti-inflammatory drugs (NSAIDs). Firstly, (+)- and (-)-zaltoprofen glucuronide was biosynthesized and purified. Then a simple and rapid RP-HPLC analysis method for direct determination of (+)- and (-)-zaltoprofen glucuronide in rat hepatic microsomes was developed and validated. The calibration curves of (+)- and (-)-zaltoprofen glucuronide both showed good linearity in the concentration range from 0.15 to 31.13 μM. The lower limit of quantification was 0.15 μM. Finally, this method was used to investigate the enantioselectivity of zaltoprofen glucuronidation in rat hepatic microsomes. The kinetics of zaltoprofen glucuronidation in rat hepatic microsomes for 40 min incubation fit the Michaelis-Menten model. Kinetic analysis indicated that (-)-zaltoprofen had a higher glucuronidation rate in rat liver microsome than that of (+)-zaltoprofen. The catalyzing efficiency (V(max)/K(m)) ratio of (+)-zaltoprofen to (-)-enantiomer is 0.8 times in rat liver microsomes.

  18. Quality assessment of Cinnamomi Ramulus by the simultaneous analysis of multiple active components using high-performance thin-layer chromatography and high-performance liquid chromatography.

    Science.gov (United States)

    Wu, Xiaoxue; He, Jiao; Xu, Huarong; Bi, Kaishun; Li, Qing

    2014-09-01

    A novel and improved method for the quality assessment of Cinnamomi Ramulus was developed and completely validated. The method was established using fingerprint technology and simultaneous quantitative determination of six main marker compounds including coumarin, cinnamic alcohol, cinnamic acid, 2-methoxy cinnamic acid, cinnamaldehyde, and 2-methoxy cinnamaldehyde in the herbal medicine for the first time. A newly developed high-performance thin-layer chromatography method, which achieved simultaneous definition of five marker components by comparing the colors and retardation factor values of the bands in high-performance thin-layer chromatography, was first used for the authentication of Cinnamomi Ramulus. The fingerprints of 26 batches of herbal samples from different regions of China showed very similar chromatographic patterns that were evaluated by similarity analysis and hierarchical clustering analysis. In addition, six marker compounds were simultaneously determined using single standard to determine multiple components by the relative response factors. Compared with the external standard method, the new quantitative method was validated to determine multiple compounds in 26 batches of Cinnamomi Ramulus samples. All results demonstrated that the simple and rapid method could be effectively utilized for the quality control of Cinnamomi Ramulus. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Integrative proteomic and microRNA analysis of the priming phase during rat liver regeneration.

    Science.gov (United States)

    Geng, Xiaofang; Chang, Cuifang; Zang, Xiayan; Sun, Jingyan; Li, Pengfei; Guo, Jianli; Xu, Cunshuan

    2016-01-10

    The partial hepatectomy (PH) model provides an effective medium for study of liver regeneration (LR). Considering that LR is regulated by microRNAs (miRNAs), investigation of the regulatory role of miRNAs is critical for revealing how regenerative processes are initiated and controlled. Using high-throughput sequencing technology, we examined miRNA expression profiles of the regenerating rat liver after PH, and found that 23 miRNAs were related to rat LR. Among them, several miRNAs were significantly altered at 2h and 6h after PH, corresponding to the priming phase of LR. Furthermore, we examined the protein profiles in the regenerating rat liver at 2h and 6h after PH by iTRAQ coupled with LC-MS/MS, and found that 278 proteins were significantly changed. Subsequently, an integrative proteomic and microRNA analysis by Ingenuity Pathway Analysis 9.0 (IPA) software showed that miR-125a, miR-143, miR-150, miR-181c, miR-182, miR-183, miR-199a, miR-429 regulated the priming phase of rat LR by modulating the expression of proteins involved in networks critical for cell apoptosis, cell survival, cell cycle, inflammatory response, metabolism, etc. Thus, our studies provide novel evidence for a functional molecular network populated by the down-regulated targets of the up-regulated miRNAs in the priming phase of rat LR.

  20. Simultaneously Excitatory and Inhibitory Effects of Transcranial Alternating Current Stimulation Revealed Using Selective Pulse-Train Stimulation in the Rat Motor Cortex.

    Science.gov (United States)

    Khatoun, Ahmad; Asamoah, Boateng; Mc Laughlin, Myles

    2017-09-27

    Transcranial alternating current stimulation (tACS) uses sinusoidal, subthreshold, electric fields to modulate cortical processing. Cortical processing depends on a fine balance between excitation and inhibition and tACS acts on both excitatory and inhibitory cortical neurons. Given this, it is not clear whether tACS should increase or decrease cortical excitability. We investigated this using transcranial current stimulation of the rat (all males) motor cortex consisting of a continuous subthreshold sine wave with short bursts of suprathreshold pulse-trains inserted at different phases to probe cortical excitability. We found that when a low-rate, long-duration, suprathreshold pulse-train was used, subthreshold cathodal tACS decreased cortical excitability and anodal tACS increased excitability. However, when a high-rate, short-duration, suprathreshold pulse-train was used this pattern was inverted. An integrate-and-fire model incorporating biophysical differences between cortical excitatory and inhibitory neurons could predict the experimental data and helped interpret these results. The model indicated that low-rate suprathreshold pulse-trains preferentially stimulate excitatory cortical neurons, whereas high-rate suprathreshold pulse-trains stimulate both excitatory and inhibitory neurons. If correct, this indicates that suprathreshold pulse-train stimulation may be able to selectively control the excitation-inhibition balance within a cortical network. The excitation-inhibition balance then likely plays an important role in determining whether subthreshold tACS will increase or decrease cortical excitability.SIGNIFICANCE STATEMENT Transcranial alternating current stimulation (tACS) is a noninvasive neuromodulation method that uses weak sinusoidal electric fields to modulate cortical activity. In healthy volunteers tACS can modulate perception, cognition, and motor function but the underlying neural mechanism is poorly understood. In this study, using rat motor

  1. Proteomic analysis of rat cerebral cortex following subchronic acrolein toxicity

    Energy Technology Data Exchange (ETDEWEB)

    Rashedinia, Marzieh; Lari, Parisa [Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of); Abnous, Khalil, E-mail: Abnouskh@mums.ac.r [Pharmaceutical Research Center, Department of Medicinal Chemistry, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of); Hosseinzadeh, Hossein, E-mail: Hosseinzadehh@mums.ac.ir [Pharmaceutical Research Center, Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of)

    2013-10-01

    Acrolein, a member of reactive α,β-unsaturated aldehydes, is a major environmental pollutant. Acrolein is also produced endogenously as a toxic by-product of lipid peroxidation. Because of high reactivity, acrolein may mediate oxidative damages to cells and tissues. It has been shown to be involved in a wide variety of pathological states including pulmonary, atherosclerosis and neurodegenerative diseases. In this study we employed proteomics approach to investigate the effects of subchronic oral exposures to 3 mg/kg of acrolein on protein expression profile in the brain of rats. Moreover effects of acrolein on malondialdehyde (MDA) levels and reduced glutathione (GSH) content were investigated. Our results revealed that treatment with acrolein changed levels of several proteins in diverse physiological process including energy metabolism, cell communication and transport, response to stimulus and metabolic process. Interestingly, several differentially over-expressed proteins, including β-synuclein, enolase and calcineurin, are known to be associated with human neurodegenerative diseases. Changes in the levels of some proteins were confirmed by Western blot. Moreover, acrolein increases the level of MDA, as a lipid peroxidation biomarker and decreased GSH concentrations, as a non-enzyme antioxidant in the brain of acrolein treated rats. These findings suggested that acrolein induces the oxidative stress and lipid peroxidation in the brain, and so that may contribute to the pathophysiology of neurological disorders. - Highlights: • Acrolein intoxication increased lipid peroxidation and deplete GSH in rat brain. • Effect of acrolein on protein levels of cerebral cortex was analyzed by 2DE-PAGE. • Levels of a number of proteins with different biological functions were increased.

  2. Simultaneous determination of zinc, cadmium and lead in environmental water samples by potentiometric stripping analysis (PSA) using multiwalled carbon nanotube electrode

    Energy Technology Data Exchange (ETDEWEB)

    Teixeira Tarley, Cesar Ricardo, E-mail: ctarleyquim@yahoo.com.br [Departamento de Ciencias Exatas, Universidade Federal de Alfenas, Unifal-MG, Alfenas, MG 37130-000 (Brazil); Silva Santos, Vivian; Lobo Baeta, Bruno Eduardo [Departamento de Ciencias Exatas, Universidade Federal de Alfenas, Unifal-MG, Alfenas, MG 37130-000 (Brazil); Cesar Pereira, Arnaldo; Tatsuo Kubota, Lauro [Instituto de Quimica, Departamento de Quimica Analitica, Universidade Estadual de Campinas, Unicamp, Campinas, SP 13083-970 (Brazil)

    2009-09-30

    The present paper has focused on the potential application of multiwalled carbon nanotube for the development of a new, simple and highly selective electrochemical method for simultaneous Zn (II), Cd (II) and Pb (II) monitoring in water samples (lake and effluent waters), by using potentiometric stripping analysis (PSA). The electrochemical method is based on simultaneous preconcentration/reduction of metal ions onto a multiwall carbon nanotube electrode at -1.3 V (versus Ag/AgCl{sub sat}) in 0.3 mol L{sup -1} acetate solution containing 15 mg L{sup -1} Hg (II) ions during 180 s, followed by subsequent chemical stripping. The analytical curve for all analytes covered the linear range varying from 58.4 up to 646.2 {mu}g L{sup -1} with correlation coefficients higher than 0.981. The limits of detection for Zn (II), Cd (II) and Pb (II) were found to be 28.0, 8.4 and 6.6 {mu}g L{sup -1}, while the relative standard deviation (RSD) at 352 {mu}g L{sup -1} was 5.6, 7.1 and 5.6% (n = 5), respectively. The behavior of the simultaneous determination in the presence of following ions Co (II), Cr (III) and Cu (II) was affected by using the analyte:interferent ratio 1:10. Therefore, by using standard addition method, Zn (II), Cd (II) and Pb (II) ions in lake and effluent water samples were determined after the spiking procedure and the results were successfully compared with those obtained by atomic absorption spectrometry (AAS). The obtained results suggest that the proposed method can be applied as a simple and efficient alternative for the simultaneous monitoring of heavy metals in water samples, according to those established requirements from environmental organizations. In addition, this method demonstrates the powerful application of carbon nanotubes in the field of potentiometric stripping analysis.

  3. OBJECTIVE EVALUATION OF HYPERACTIVATED MOTILITY IN RAT SPERMATOZA USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

    Science.gov (United States)

    Objective evaluation of hyperactivated motility in rat spermatozoa using computer-assisted sperm analysis.Cancel AM, Lobdell D, Mendola P, Perreault SD.Toxicology Program, University of North Carolina, Chapel Hill, NC 27599, USA.The aim of this study was t...

  4. Pixe analysis of trace elements in tissues of rats treated with anticonvulsants

    Science.gov (United States)

    Hurd, R. W.; Van Rinsvelt, H. A.; Kinyua, A. M.; O'Neill, M. P.; Wilder, B. J.; Houdayer, A.; Hinrichsen, P. F.

    1987-04-01

    Several lines of evidence implicate metals in epilepsy. Anticonvulsant drugs are noted to alter levels of metals in humans and animals. PIXE analysis was used to investigate effects of three anticonvulsant drugs on tissue and brain cortex trace elements. The content of zinc and copper was increased in liver and spleen of rats treated with anticonvulsants while selenium was decreased in cortex.

  5. PIXE analysis of trace elements in tissues of rats treated with anticonvulsants

    Energy Technology Data Exchange (ETDEWEB)

    Hurd, R.W.; Van Rinsvelt, H.A.; Kinyua, A.M.; O' Neill, M.P.; Wilder, B.J.; Houdayer, A.; Hinrichsen, P.F.

    1987-04-01

    Several lines of evidence implicate metals in epilepsy. Anticonvulsant drugs are noted to alter levels of metals in humans and animals. PIXE analysis was used to investigate effects of three anticonvulsant drugs on tissue and brain cortex trace elements. The content of zinc and copper was increased in liver and spleen of rats treated with anticonvulsants while selenium was decreased in cortex.

  6. OBJECTIVE EVALUATION OF HYPERACTIVATED MOTILITY IN RAT SPERMATOZA USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

    Science.gov (United States)

    Objective evaluation of hyperactivated motility in rat spermatozoa using computer-assisted sperm analysis.Cancel AM, Lobdell D, Mendola P, Perreault SD.Toxicology Program, University of North Carolina, Chapel Hill, NC 27599, USA.The aim of this study was t...

  7. Combined sequence-based and genetic mapping analysis of complex traits in outbred rats

    NARCIS (Netherlands)

    Baud, Amelie; Hermsen, Roel; Guryev, Victor; Stridh, Pernilla; Graham, Delyth; McBride, Martin W.; Foroud, Tatiana; Calderari, Sophie; Diez, Margarita; Ockinger, Johan; Beyeen, Amennai D.; Gillett, Alan; Abdelmagid, Nada; Guerreiro-Cacais, Andre Ortlieb; Jagodic, Maja; Tuncel, Jonatan; Norin, Ulrika; Beattie, Elisabeth; Huynh, Ngan; Miller, William H.; Koller, Daniel L.; Alam, Imranul; Falak, Samreen; Osborne-Pellegrin, Mary; Martinez-Membrives, Esther; Canete, Toni; Blazquez, Gloria; Vicens-Costa, Elia; Mont-Cardona, Carme; Diaz-Moran, Sira; Tobena, Adolf; Hummel, Oliver; Zelenika, Diana; Saar, Kathrin; Patone, Giannino; Bauerfeind, Anja; Bihoreau, Marie-Therese; Heinig, Matthias; Lee, Young-Ae; Rintisch, Carola; Schulz, Herbert; Wheeler, David A.; Worley, Kim C.; Muzny, Donna M.; Gibbs, Richard A.; Lathrop, Mark; Lansu, Nico; Toonen, Pim; Ruzius, Frans Paul; de Bruijn, Ewart; Hauser, Heidi; Adams, David J.; Keane, Thomas; Atanur, Santosh S.; Aitman, Tim J.; Flicek, Paul; Malinauskas, Tomas; Jones, E. Yvonne; Ekman, Diana; Lopez-Aumatell, Regina; Dominiczak, Anna F.; Johannesson, Martina; Holmdahl, Rikard; Olsson, Tomas; Gauguier, Dominique; Hubner, Norbert; Fernandez-Teruel, Alberto; Cuppen, Edwin; Mott, Richard; Flint, Jonathan

    2013-01-01

    Genetic mapping on fully sequenced individuals is transforming understanding of the relationship between molecular variation and variation in complex traits. Here we report a combined sequence and genetic mapping analysis in outbred rats that maps 355 quantitative trait loci for 122 phenotypes. We i

  8. CLONING, EXPRESSION, AND MUTATIONAL ANALYSIS OF RAT S-ADENOSYL-1-METHIONINE: ARSENIC (III) METHYLTRANSFERASE

    Science.gov (United States)

    CLONING, EXPRESSION, AND MUTATIONAL ANALYSIS OF RAT S-ADENOSYL-L-METHIONINE: ARSENIC(III) METHYLTRANSFERASEStephen B. Waters, Ph.D., Miroslav Styblo, Ph.D., Melinda A. Beck, Ph.D., University of North Carolina at Chapel Hill; David J. Thomas, Ph.D., U.S. Environmental...

  9. Simultaneous central nervous system distribution and pharmacokinetic–pharmacodynamic modelling of the electroencephalogram effect of norfloxacin administered at a convulsant dose in rats

    Science.gov (United States)

    Chenel, Marylore; Marchand, Sandrine; Dupuis, Antoine; Lamarche, Isabelle; Paquereau, Joël; Pariat, Claudine; Couet, William

    2004-01-01

    The objective of this study was to investigate the contribution of norfloxacin blood–brain barrier (BBB) transport to its delayed electroencephalogram (EEG) effect in rats. Norfloxacin was injected as a bolus dose of 150 mg kg−1. Blood samples were collected for total norfloxacin plasma concentration measurements. The corresponding unbound levels were determined in brain extracellular fluid (ECF) using microdialysis. Quantitative EEG recording was conducted during 9 h post-dose. Brain ECF norfloxacin concentrations were much lower than plasma levels (AUC ratio=9.7±2.8%) but peaked very early, and concentration versus time profiles were parallel in both biological fluids. The best pharmacokinetic (PK) modelling was obtained by considering that ECF concentrations were part of the central compartment, with a proportionality factor. The peak of EEG effect was delayed and the effect versus plasma concentration curves exhibited a dramatic hysteresis. A PK–pharmacodynamic (PD) effect compartment model with a spline function to describe the relationship between effect and concentration at the effect site successfully described the data. Comparisons of PK–PD parameters estimated from plasma and ECF concentrations show that most of the delayed norfloxacin EEG effect is not due to BBB transport, but also that PD parameters derived from plasma data must be carefully interpreted when drug distribution at the effect site is restricted, as may often be the case for centrally acting drugs. PMID:15155539

  10. Simultaneous Voltammetric Measurements of Glucose and Dopamine Demonstrate the Coupling of Glucose Availability with Increased Metabolic Demand in the Rat Striatum.

    Science.gov (United States)

    Smith, Samantha K; Lee, Christie A; Dausch, Matthew E; Horman, Brian M; Patisaul, Heather B; McCarty, Gregory S; Sombers, Leslie A

    2017-02-15

    Cerebral blood flow ensures delivery of nutrients, such as glucose, to brain sites with increased metabolic demand. However, little is known about rapid glucose dynamics at discrete locations during neuronal activation in vivo. Acute exposure to many substances of abuse elicits dopamine release and neuronal activation in the striatum; however, the concomitant changes in striatal glucose remain largely unknown. Recent developments have combined fast-scan cyclic voltammetry with glucose oxidase enzyme modified carbon-fiber microelectrodes to enable the measurement of glucose dynamics with subsecond temporal resolution in the mammalian brain. This work evaluates several waveforms to enable the first simultaneous detection of endogenous glucose and dopamine at single recording sites. These molecules, one electroactive and one nonelectroactive, were found to fluctuate in the dorsal striatum in response to electrical stimulation of the midbrain and systemic infusion of cocaine/raclopride. The data reveal the second-by-second dynamics of these species in a striatal microenvironment, and directly demonstrate the coupling of glucose availability with increased metabolic demand. This work provides a foundation that will enable detailed investigation of local mechanisms that regulate the coupling of cerebral blood flow with metabolic demand under normal conditions, and in animal studies of drug abuse and addiction.

  11. Theoretical Analysis on the Simultaneous Operation of a Color-Center Laser and a Q-Switched Nd: YAG Laser

    Institute of Scientific and Technical Information of China (English)

    WU Xing; DING Yongkui; ZHANG Rui

    2000-01-01

    The simultaneous operation for the Q- switched Nd: YAG laser at 1.06 and the tunable color-center laser over 1.12~1.26 μm has been realized by using a LiF: F2- color-center crystal both as the Q-switcher for the Nd:YAG laser and as the active medium for the color-center laser. The interaction of the two lasers has been analyzed and calculated with the rate equations. The pulse duration of YAG laser is compressed, in agreement with the experimental results.

  12. Simultaneous high performance liquid chromatographic analysis of vitamins B1, B2 and B6 in royal jelly

    OpenAIRE

    2004-01-01

    Royal jelly is used as a food supplement, popularly known as rich in B vitamins. The present work has two objectives: firstly, to apply simultaneous quantitative determination by High Performance Liquid Chromatography of thiamin (vitamin B1), riboflavin (vitamin B2) and pyridoxine (vitamin B6) and secondly to compare the obtained data with the Dietary Reference Intake (DRI) values. The values obtained showed no thiamin, a range from 20 to 171 ng g-1 of riboflavin and from 408 to 2 188 ng g-1 ...

  13. Proteomic analysis of rat cerebral cortex following subchronic acrolein toxicity.

    Science.gov (United States)

    Rashedinia, Marzieh; Lari, Parisa; Abnous, Khalil; Hosseinzadeh, Hossein

    2013-10-01

    Acrolein, a member of reactive α,β-unsaturated aldehydes, is a major environmental pollutant. Acrolein is also produced endogenously as a toxic by-product of lipid peroxidation. Because of high reactivity, acrolein may mediate oxidative damages to cells and tissues. It has been shown to be involved in a wide variety of pathological states including pulmonary, atherosclerosis and neurodegenerative diseases. In this study we employed proteomics approach to investigate the effects of subchronic oral exposures to 3mg/kg of acrolein on protein expression profile in the brain of rats. Moreover effects of acrolein on malondialdehyde (MDA) levels and reduced glutathione (GSH) content were investigated. Our results revealed that treatment with acrolein changed levels of several proteins in diverse physiological process including energy metabolism, cell communication and transport, response to stimulus and metabolic process. Interestingly, several differentially over-expressed proteins, including β-synuclein, enolase and calcineurin, are known to be associated with human neurodegenerative diseases. Changes in the levels of some proteins were confirmed by Western blot. Moreover, acrolein increases the level of MDA, as a lipid peroxidation biomarker and decreased GSH concentrations, as a non-enzyme antioxidant in the brain of acrolein treated rats. These findings suggested that acrolein induces the oxidative stress and lipid peroxidation in the brain, and so that may contribute to the pathophysiology of neurological disorders.

  14. Simultaneous determination of puerarin, daidzin, daidzein, paeoniflorin, albiflorin, liquiritin and liquiritigenin in rat plasma and its application to a pharmacokinetic study of Ge-Gen Decoction by a liquid chromatography-electrospray ionization-tandem mass spectrometry.

    Science.gov (United States)

    Yan, Yan; Chai, Cheng-Zhi; Wang, Da-Wei; Wu, Jie; Xiao, Hong-He; Huo, Li-Xia; Zhu, Dan-Ni; Yu, Bo-Yang

    2014-07-01

    A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for simultaneous determination of seven constituents including puerarin, daidzin, daidzein, paeoniflorin, albiflorin, liquiritin and liquiritigenin in rat plasma using schisandrin as the internal standard (IS). The plasma samples were pretreated by a one-step direct protein precipitation with acetonitrile. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and water (containing 0.1% formic acid and 5mM ammonium acetate). All analytes and IS were quantitated through electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. The mass transitions were as follows: m/z 417.5→297.2 for puerarin, m/z 417.1→255.2 for daidzin, m/z 255.2→152.4 for daidzein, m/z 498.1→179.3 for paeoniflorin, m/z 481.1→197.3 for albiflorin, m/z 436.2→257.3 for liquiritin, m/z 257.2→137.3 for liquiritigenin and m/z 415.0→384.2 for IS, respectively. All calibration curves exhibited good linearity (r>0.9979) over a wide concentration range for all components. The intra-day and inter-day precisions (RSD) at three different levels were both less than 14.3% and the accuracies (RE) ranged from -13.2% to 14.8%. The extraction recoveries of the seven compounds ranged from 72.9% to 117.4%. The validated method was successfully applied to pharmacokinetic study of the seven components in female rat plasma after oral administration of Ge-Gen Decoction aqueous extract.

  15. Ultra-performance liquid chromatography-tandem mass spectrometric assay for the simultaneous determination of brucine, strychnine and brucine N-oxide in rat plasma: application to a pharmacokinetic study.

    Science.gov (United States)

    Gu, Wei; Wang, Dongyue; Pan, Zihao; Liu, Xiao; Cai, Baochang; Chen, Jun

    2016-07-01

    A rapid, simple and sensitive UHPLC-MS/MS method was developed and validated for the simultaneous determination of brucine, strychnine and brucine N-oxide in rat plasma using huperzine A as an internal standard (IS) after protein precipitation with methanol. The analytes were separated on a Purospher® STAR RP18 UHPLC column (2 µm, 2.1 × 100 mm) by gradient elution using a mobile phase composed of methanol and water (containing 0.1% formic acid) at a flow rate of 0.3 mL/min. Brucine, strychnine, brucine N-oxide and IS were detected in positive ion multiple reaction monitoring mode by means of an electrospray ionization interface (m/z 395.2 → 324.1, m/z 335.2 → 184.1, m/z 411.2 → 394.2, m/z 243.1 → 226.1). The calibration curve was linear over the range of 1-500 ng/mL for brucine and strychnine and 0.2-50 ng/mL for brucine N-oxide. The intra- and inter-day precisions of these analytes were all within 15% and the accuracy ranged from 85 to 115%. The stability experiment indicated that the plasma samples at three concentration levels were stable under different conditions. The developed method was successfully applied for the first time to pharmacokinetic studies of brucine, strychnine and brucine N-oxide following a single oral and intravenous administration of modified total alkaloid fraction in rats. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  16. A UPLC-MS/MS method for simultaneous quantitation of three monoterpene glycosides and four alkaloids in rat plasma: application to a comparative pharmacokinetic study of Huo Luo Xiao Ling Dan and single herb extract.

    Science.gov (United States)

    Ai, Yu; Wu, Yun; Wang, Fenrong; Ma, Wen; Bian, Qiaoxia; Lee, David Y-W; Dai, Ronghua

    2015-03-01

    The objective of this study was to develop a sensitive and reliable ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous quantitation of three monoterpene glycosides (paeoniflorin, alibiflorin and oxypaeoniflorin) and four alkaloids (tetrahydropalmatine, corydaline, dehydrocorydaline and berberine), the main active ingredients of Radix Paeoniae Rubra extract (RPE) and Corydalis yanhusuo extract (CYE) in Huo Luo Xiao Ling Dan (HLXLD), and to compare the pharmacokinetics of these active ingredients in normal and arthritic rats orally administrated with HLXLD or RPE/CYE alone. The analytes and internal standard (IS) (geniposide) were separated on a XBridge C18 column (150 × 4.6 mm, 3.5 µm) using gradient elution with the mobile phase consisting of methanol and 0.01% formic acid in water at a flow rate of 0.6 ml/min. The detection of the analytes was performed on Acquity UPLC-MS/MS system with an electrospray ionization and multiple reaction monitoring mode via polarity switching between negative (for monoterpene glycosides) and positive (for alkaloids) ionization mode. The lower limits of quantification were 2.5, 1, 0.5, 0.2, 0.2, 0.02 and 0.01 ng/ml for paeoniflorin, alibiflorin, oxypaeoniflorin, tetrahydropalmatine, corydaline, dehydrocorydaline and berberine, respectively. Intra-day and inter-day precision and accuracy of analytes were well within acceptance criteria (15%). The mean extraction recoveries of analytes and IS from rat plasma were all more than 83.1%. The validated method has been successfully applied to determination of the analytes. Results showed that there were remarkable differences in pharmacokinetic properties of the analytes between herbal formula and single herb group, normal and arthritic group.

  17. A UHPLC-MS/MS method for simultaneous determination of six flavonoids, gallic acid and 5,8-dihydroxy-1,4-naphthoquinone in rat plasma and its application to a pharmacokinetic study of Cortex Juglandis Mandshuricae extract.

    Science.gov (United States)

    Sun, Zhi; Zhao, Longshan; Zuo, Lihua; Qi, Chao; Zhao, Pan; Hou, Xiaohong

    2014-05-01

    Cortex Juglandis Mandshuricae is used as a folk remedy for treating cancer, diarrhea and dysentery in traditional Chinese medicine for many years. Six flavonoids (myricitrin, quercitrin, taxifolin, myricetin, quercetin and naringenin), gallic acid and 5,8-dihydroxy-1,4-naphthoquinone are major bioactive components in Cortex Juglandis Mandshuricae extract. In this study, an ultrahigh performance liquid chromatography and tandem mass spectrometry method was developed for simultaneous determination of eight ingredients in rat plasma using chloromycetin as an internal standard. Plasma samples added vitamin C (antioxygen) were acidified with hydrochloric acid and extracted by liquid-liquid extraction with ethyl acetate. Eight ingredients were separated on a Venusil ASB C18 column and detected by multiple reaction monitoring mode using electrospray ionization in the negative ion mode. The method was linear for all analytes over investigated range with all correlation coefficients greater than 0.9900. The validated lower limit of quantification was 20ng/mL for gallic acid, 5ng/mL for myricitrin, 3ng/mL for quercitrin, 10ng/mL for taxifolin, 6ng/mL for myricetin, 3ng/mL for quercetin, 2ng/mL for naringenin and 1μg/mL for 5,8-dihydroxy-1,4-naphthoquinone, respectively. Intra- and inter-day precisions (RSD%) were less than 15% and accuracy (RE%) ranged from -6.9% to 6.9%. The validated method was successfully applied to investigate the pharmacokinetics of the eight analytes after oral administration of Cortex Juglandis Mandshuricae extract to rats.

  18. Development and validation of an LC/MS/MS method for simultaneous determination of shionone and epi-friedelinol in rat plasma for pharmacokinetic study after oral administration of Aster tataricus extract.

    Science.gov (United States)

    Yin, De-Feng; Zhou, Kai; Liu, Ji-Tao; Hu, Li; Liu, Ying; Deng, Jun; Wang, Song-Ping; Xiong, Ying; Zhong, Wu

    2016-07-01

    A rapid and sensitive liquid chromatography tandem mass spectrometry (LC/MS/MS) method was developed and validated using spinasterol as the internal standard (IS) for the simultaneous determination of shionone and epi-friedelinol in rat plasma. Plasma samples were pretreated using liquid-liquid extraction with ethyl ether. Chromatographic separation was achieved on a C18 column (100 × 2.1 mm, 5 μm) with an isocratic elution consisting of acetonitrile-0.1% formic acid water (75:25, v/v) at a flow rate of 0.30 mL/min. Detection was performed under the selected reaction monitoring scan using an electrospray ionization in the positive ion mode. The mass transitions were as follows: m/z 427.4 → 95.1 for shionone, m/z 411.4 → 205.2 for epi-friedelinol and m/z 395.3 → 105.2 for IS. All calibration curves exhibited good linearity (r > 0.995) over the concentration range for both components. The intra- and inter-day precisions at three QC and lower limit of quantitation levels were both extraction recoveries of the compounds ranged from 82.07 to 89.81%. The developed method was successfully applied to the pharmacokinetic study of shionone and epi-friedelinol after oral administration of Aster tataricus extract to rats. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Development of a LC-MS/MS method for simultaneous determination of metoprolol and its metabolites, α-hydroxymetoprolol and O-desmethylmetoprolol, in rat plasma: application to the herb-drug interaction study of metoprolol and breviscapine.

    Science.gov (United States)

    Rao, Zhi; Ma, Yan-rong; Qin, Hong-yan; Wang, Ya-feng; Wei, Yu-hui; Zhou, Yan; Zhang, Guo-qiang; Wang, Xing-dong; Wu, Xin-an

    2015-09-01

    A simple, specific and sensitive LC-MS/MS method was developed and validated for the simultaneous determination of metoprolol (MET), α-hydroxymetoprolol (HMT) and O-desmethylmetoprolol (DMT) in rat plasma. The plasma samples were prepared by protein precipitation, then the separation of the analytes was performed on an Agilent HC-C18 column (4.6 × 250 mm, 5 µm) at a flow rate of 1.0 mL/min, and post-column splitting (1:4) was used to give optimal interface flow rates (0.2 mL/min) for MS detection; the total run time was 8.5 min. Mass spectrometric detection was achieved using a triple-quadrupole mass spectrometer equipped with an electrospray source interface in positive ionization mode. The method was fully validated in terms of selectivity, linearity, accuracy, precision, stability, matrix effect and recovery over a concentration range of 3.42-7000 ng/mL for MET, 2.05-4200 ng/mL for HMT and 1.95-4000 ng/mL for DMT. The analytical method was successfully applied to herb-drug interaction study of MET and breviscapine after administration of breviscapine (12.5 mg/kg) and MET (40 mg/kg). The results suggested that breviscapine have negligible effect on pharmacokinetics of MET in rats; the information may be beneficial for the application of breviscapine in combination with MET in clinical therapy.

  20. Rat sciatic nerve crush injury and recovery tracked by plantar test and immunohistochemistry analysis.

    Science.gov (United States)

    Pavić, Roman; Pavić, Michele L; Tvrdeić, Ante; Tot, Ozana K; Heffer, Marija

    2011-01-01

    An experimental crush injury to the sciatic nerve, with a crush force of 49.2 N (pressure p=1.98x10(8) Pa), was inflicted in 30 male rats (Wistar). A control group (sham), with the same number of rats, was also operated upon exactly as the experimental group but without the crush injury. We tested the sensory and motor recovery of the sciatic nerve with Hargreaves method, using an apparatus from Ugo Basile, Italy. Testing was continued for both legs of each rat, injured and uninjured, starting preoperatively (0 day), and then 1, 7, 14, 21, and 28 days postoperatively. The same experiment was run simultaneously with the sham group. The Plantar test showed recovery of the sensory and motor function of the sciatic nerve, though not complete recovery, by 28 days. An immunohistochemical experiment was run in parallel with the plantar test on L3-L6 segments of the spinal cord from where the sciatic nerve extends. We used antibodies for Myelin-associated glycoprotein (MAG), and gangliosides GD1a and GT1b on the aforesaid part of the spinal cord. The immunohistochemical methods showed changes in sensory and motor axons in the spinal cord segment L3-L6 which suggest correspondence with the results of the Plantar test, in terms of recovery of the sensory and motor function after injury of the sciatic nerve. The immunohistochemical results also show ipsilateral and contralateral changes following injury. Results of the plantar test are suggestive that the rat shows compensation for an injury in its contralateral leg.

  1. Proteomic Profiling Analysis Reveals a Link between Experimental Autoimmune Uveitis and Complement Activation in Rats.

    Science.gov (United States)

    Guo, D D; Hu, B; Tang, H Y; Sun, Y Y; Liu, B; Tian, Q M; Bi, H S

    2017-05-01

    Uveitis is an autoimmune disease that usually damages the vision function, leading to poor visual quality in patients. As an autoimmune ocular inflammatory disease, the pathogenesis of uveitis is associated with abnormal expression of some proteins and aberrant regulation of multiple signalling pathways. Nevertheless, the detailed mechanism remains unclear. In this study, we induced an experimental autoimmune uveitis (EAU) model in rats. We determined the levels of C3a and membrane attack complex C5b-9 (soluble C5b-9, sC5b-9) in both plasma and aqueous humour, identified the differentially expressed proteins in plasma by liquid chromatography-tandem mass spectrometry and employed bioinformatics algorithms to analyse differentially expressed proteins in EAU rat plasma. The results demonstrate that there were 168 differentially expressed plasma proteins in EAU rats versus control subjects. The levels of sC5b-9 and C3a were elevated in the plasmas and aqueous humours of EAU rats. Gene ontology enrichment analysis showed that the differentially expressed proteins in EAU rat plasma were mainly involved in metabolic and immune processes. Kyoto encyclopedia of genes and genomes (KEGG) pathway annotation, database for annotation, visualization and integrated discovery (DAVID) and protein-protein interaction analyses revealed that the differentially expressed proteins in EAU rat plasmas were closely associated with complement and coagulation cascades, metabolic pathways, NF-kappa B, PI3K-Akt, Toll-like receptors and autophagy. Overall, the differentially expressed proteins in EAU rat plasmas are mainly involved in the complement and coagulation cascades. The pathogenesis of uveitis closely correlates with complement activation. © 2017 The Foundation for the Scandinavian Journal of Immunology.

  2. Zinc fixation preserves flow cytometry scatter and fluorescence parameters and allows simultaneous analysis of DNA content and synthesis, and intracellular and surface epitopes

    DEFF Research Database (Denmark)

    Jensen, Uffe Birk; Owens, David; Pedersen, Søren

    2010-01-01

    Zinc salt-based fixation (ZBF) has proved advantageous in histochemical analyses conducted on intact tissues but has not been exploited in flow cytometry procedures that focus on quantitative analysis of individual cells. Here, we show that ZBF performs equally well to paraformaldehyde in the pre......Zinc salt-based fixation (ZBF) has proved advantageous in histochemical analyses conducted on intact tissues but has not been exploited in flow cytometry procedures that focus on quantitative analysis of individual cells. Here, we show that ZBF performs equally well to paraformaldehyde...... allowing subsequent quantitative PCR analysis or labeling for incorporation of the thymidine analog EdU following surface and intracellular epitope staining. Finally, ZBF treatment allows for long-term storage of labeled cells with little change in these parameters. Thus, we present a protocol for zinc...... salt fixation of cells that allows for the simultaneous analysis of DNA and intracellular and cell surface proteins by flow cytometry....

  3. Preclinical analysis of nonsteroidal anti-inflammatory drug usefulness for the simultaneous prevention of steatohepatitis, atherosclerosis and hyperlipidemia

    Science.gov (United States)

    Madrigal-Perez, Violeta M; García-Rivera, Alejandro; Rodriguez-Hernandez, Alejandrina; Ceja-Espiritu, Gabriel; Briseño-Gomez, Xochitl G; Galvan-Salazar, Hector R; Soriano-Hernandez, Alejandro D; Guzman-Esquivel, Jose; Martinez-Fierro, Margarita L; Newton-Sanchez, Oscar A; Buenrostro, Bertha A Olmedo; Rodriguez-Sanchez, Iram P; López-Lemus, Uriel A; Lara-Esqueda, Agustin; Delgado-Enciso, Ivan

    2015-01-01

    Nonalcoholic steatohepatitis (NASH) is currently one of the primary liver diseases. Recent studies have shown a clinical relation between NASH and atherosclerosis. There is much interest in these two diseases because they are both associated with great morbidity and mortality. Inflammation and the overexpression of COX-2 participate in the pathophysiology of the two diseases, and therefore simultaneous treatment is feasible. The role of the four NSAIDs, meclofenamate, mefenamate, flufenamate, and aspirin, was analyzed in a mouse model of NASH, as well as preclinical atherosclerosis induced by a high-fat diet (HFD). Six mouse groups were formed. Five of the groups were fed a high-fat diet for 6 months and one group was fed a standard diet, acting as the normality reference. Of the five groups fed a high-fat diet, four received a NSAID, each of them identified by the specific drug administered. One group received no treatment. Serum markers (cholesterol, triglycerides, ALT, and AST) and histologic changes in the aorta and liver were analyzed for the study. Aspirin significantly reduced the hepaticsteatosis. All the drugs significantly reduced the hepatic inflammatory infiltrate. In relation to atherosclerosis, there were significant reductions in all the study variables with the use of aspirin and flufenamate. The four medications were able to stop steatosis from progressing into steatohepatitis by reducing inflammation. However, aspirin was the most beneficial, simultaneously reducing steatosis, atherosclerosis, and serum cholesterol levels. PMID:26885230

  4. [Comparative Analysis of Behavior in The Open-field Test in Wild Grey Rats (Rattus norvegicus) and in Grey Rats Subjected to Prolonged Selection for Tame And Aggressive Behavior].

    Science.gov (United States)

    Kozhemyakina, R V; Konoshenko, M Yu; Sakharov, D G; Smagin, D A; Markel, A L

    2016-01-01

    The aim of this work is analysis of the open-field behavior in grey rats selected for the tame and aggressive behavior in comparison with the wild grey rats. Significant influences of the rat group factor on the 13 of 19 behavioral features studied in the open-field were found. This effect, in general, depends on existence of great differences between behaviors of the wild rats from the one hand and behaviors of the tame and aggressive rats from the other. The behaviors of the rats from the last two groups are practically identical. Multidimensional analysis confirms the distinct separation in coordinates of the two main components of the wild rat behavior from the behavior of both the tame and selectively bred aggressive rats. The first main component dimension corresponds to the grade of fear, which was significantly enhanced in the wild rats. So, in spite of the equality of behavioral aggressiveness of the wild rats and the rats selected for aggression with the glove test, the behavior of selected aggressive rats in the open-field is analogous to behavior of the rats selected for tameness. Comparison of behavioral features with the hormonal stress responsiveness allowed us to conclude that the aggressive behavior of the wild and se lected for aggression rats based on different motivational and neuroendocrine processes.

  5. A novel multispecific competitor fragment for quantitative PCR analysis of cytokine gene expression in rats.

    Science.gov (United States)

    Siegling, A; Lehmann, M; Platzer, C; Emmrich, F; Volk, H D

    1994-12-28

    Competitive polymerase chain reaction (PCR) is a sensitive method for quantification of cytokine mRNA expression. Co-amplification of an internal standard serves as control for comparing the efficiency of PCR in different samples. We have developed a novel control fragment for multiple analyses of rat cytokine gene expression containing primers for IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, TNF-alpha, TGF-beta 1, IFN-gamma and MIP-2. Additional primers were incorporated to analyse the content of T cells (CD3), activated T cells (CD25) and housekeeping genes (beta-actin and HPRT). As an example we demonstrate analysis of IL-2 mRNA expression in small pieces of kidney tissue obtained from rats after kidney allotransplantation. The IL-2 expression decreased tenfold during treatment with an anti-rat CD4 monoclonal antibody as compared to untreated animals.

  6. Metabonomic analysis of Allium macrostemon Bunge as a treatment for acute myocardial ischemia in rats.

    Science.gov (United States)

    Li, Fang; Xu, Qian; Zheng, Ting; Huang, Fang; Han, Lintao

    2014-01-01

    Myocardial ischemia (MI) refers to a pathological state of the heart caused by reduced cardiac blood perfusion, which leads to a decreased oxygen supply in the heart and an abnormal myocardial energy metabolism. Acute myocardial ischemia (AMI) has posed a significant health risk for humans. Allium macrostemon Bunge (AMB), a popular traditional Chinese medicine, is used for MI treatment. The therapeutic effects of AMB were assessed and the detailed mechanisms of AMB for AMI treatment were investigated. We characterized the metabonomic variations in rats from the sham surgery, AMI, and AMB-pretreated AMI groups through a combination of nuclear magnetic resonance (NMR) spectroscopy and multivariate statistical analysis. Thirty-five metabolites including carbohydrates, a range of amino acids, and organic acids were detected. The (1)H NMR spectra of the rat serum were analyzed using the principal component analysis (PCA) and orthogonal projection to latent structures discriminate analysis (OPLS-DA). Results showed that AMI induced some physiological changes in rats and also led to metabolic disorders related to glycolysis promotion, amino acid metabolism disruption, and other metabolite metabolism perturbation. AMB pretreatment reduced the AMI injury and maintained metabolic balance, possibly by limiting the change in energy metabolism and regulating amino acid metabolism. These findings provide a comprehensive insight on the metabolic response of AMI rats to AMB pretreatment and are important for the use of AMB for AMI therapy.

  7. Quantitative Analysis of Micro-CT Imaging and Histopathological Signatures of Experimental Arthritis in Rats

    Directory of Open Access Journals (Sweden)

    Matthew D. Silva

    2004-10-01

    Full Text Available Micro-computed tomographic (micro-CT imaging provides a unique opportunity to capture 3-D architectural information in bone samples. In this study of pathological joint changes in a rat model of adjuvant-induced arthritis (AA, quantitative analysis of bone volume and roughness were performed by micro-CT imaging and compared with histopathology methods and paw swelling measurement. Micro-CT imaging of excised rat hind paws (n = 10 stored in formalin consisted of approximately 600 30-μm slices acquired on a 512 × 512 image matrix with isotropic resolution. Following imaging, the joints were scored from H&E stained sections for cartilage/bone erosion, pannus development, inflammation, and synovial hyperplasia. From micro-CT images, quantitative analysis of absolute bone volumes and bone roughness was performed. Bone erosion in the rat AA model is substantial, leading to a significant decline in tarsal volume (27%. The result of the custom bone roughness measurement indicated a 55% increase in surface roughness. Histological and paw volume analyses also demonstrated severe arthritic disease as compared to controls. Statistical analyses indicate correlations among bone volume, roughness, histology, and paw volume. These data demonstrate that the destructive progression of disease in a rat AA model can be quantified using 3-D micro-CT image analysis, which allows assessment of arthritic disease status and efficacy of experimental therapeutic agents.

  8. Gene expression microarray analysis of early oxygen-induced retinopathy in the rat.

    Science.gov (United States)

    Tea, Melinda; Fogarty, Rhys; Brereton, Helen M; Michael, Michael Z; Van der Hoek, Mark B; Tsykin, Anna; Coster, Douglas J; Williams, Keryn A

    2009-12-12

    Different inbred strains of rat differ in their susceptibility to oxygen-induced retinopathy (OIR), an animal model of human retinopathy of prematurity. We examined gene expression in Sprague-Dawley (susceptible) and Fischer 344 (resistant) neonatal rats after 3 days exposure to cyclic hyperoxia or room air, using Affymetrix rat Genearrays. False discovery rate analysis was used to identify differentially regulated genes. Such genes were then ranked by fold change and submitted to the online database, DAVID. The Sprague-Dawley list returned the term "response to hypoxia," absent from the Fischer 344 output. Manual analysis indicated that many genes known to be upregulated by hypoxia-inducible factor-1alpha were downregulated by cyclic hyperoxia. Quantitative real-time RT-PCR analysis of Egln3, Bnip3, Slc16a3, and Hk2 confirmed the microarray results. We conclude that combined methodologies are required for adequate dissection of the pathophysiology of strain susceptibility to OIR in the rat. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12177-009-9041-7) contains supplementary material, which is available to authorized users.

  9. PCR Conditions for 16S Primers for Analysis of Microbes in the Colon of Rats.

    Science.gov (United States)

    Guillen, I A; Camacho, H; Tuero, A D; Bacardí, D; Palenzuela, D O; Aguilera, A; Silva, J A; Estrada, R; Gell, O; Suárez, J; Ancizar, J; Brown, E; Colarte, A B; Castro, J; Novoa, L I

    2016-09-01

    The study of the composition of the intestinal flora is important to the health of the host, playing a key role in maintaining intestinal homeostasis and the evolution of the immune system. For these studies, various universal primers of the 16S rDNA gene are used in microbial taxonomy. Here, we report an evaluation of 5 universal primers to explore the presence of microbial DNA in colon biopsies preserved in RNAlater solution. The DNA extracted was used for the amplification of PCR products containing the variable (V) regions of the microbial 16S rDNA gene. The PCR products were studied by restriction fragment length polymorphism (RFLP) analysis and DNA sequence, whose percent of homology with microbial sequences reported in GenBank was verified using bioinformatics tools. The presence of microbes in the colon of rats was quantified by the quantitative PCR (qPCR) technique. We obtained microbial DNA from rat, useful for PCR analysis with the universal primers for the bacteria 16S rDNA. The sequences of PCR products obtained from a colon biopsy of the animal showed homology with the classes bacilli (Lactobacillus spp) and proteobacteria, normally represented in the colon of rats. The proposed methodology allowed the attainment of DNA of bacteria with the quality and integrity for use in qPCR, sequencing, and PCR-RFLP analysis. The selected universal primers provided knowledge of the abundance of microorganisms and the formation of a preliminary test of bacterial diversity in rat colon biopsies.

  10. Effect of methylprednisolone in medullar injury in rats: a functional and histological analysis

    Directory of Open Access Journals (Sweden)

    Tebet Marcos Antônio

    2003-01-01

    Full Text Available The pharmacological effects of the methylprednisolone in the treatment of the spinal cord lesion, have been researched. An experimental study was performed with twelve wistar rats subjected to a contusion lesion of the spinal cord, by the NYU system. The rats were divided in two groups; the MP group which received the methylprednisolone after the contusion lesion and the control group, which received a 0,9% physiological serum. An analysis of the recuperation of the functional movement of the rats was accomplished on the 2nd, 7th and 14th day after the contusion, utilizing the BBB test. On the fourteenth day the rats were sacrificed and the histological findings of the spinal cord lesion were analyzed. It was noted that the rats from the MP group exhibited an improvement in the recuperation of their functional movement compared to that of the control group, and the histological findings of the spinal cord lesion couldn't be correlated with the recuperation of their functional movement.

  11. Comparative biochemical analysis of sea urchin peristome and rat tail tendon collagen.

    Science.gov (United States)

    Robinson, J J

    1997-06-01

    We report here a biochemical comparison between type 1 rat tail tendon collagen and collagen isolated from sea urchin peristome tissue. The sea urchin collagen consisted of two species of apparent mol masses, 140 and 116 kDa. Amino acid compositional analysis of the 140 and 116 kDa species revealed the presence of hydroxyproline and hydroxylysine as well as a glycine content of 28.1 mol.%. In solubility experiments the rat tail tendon collagen was found to precipitate at sodium chloride concentrations between 1 and 2 M while peristome collagen remained soluble at salt concentrations as high as 4 M. Incubation of the peristome and rat tail tendon collagen preparations with a sea urchin collagenase/gelatinase resulted in cleavage o