HPLC-ANALYSIS OF ESTROGEN -ACTIVE ANABOLICA IN MEAT WITH THE ESTROGEN-RECEPTOR-TEST AS SPECIFIC DETECTION-SYSTEM

Abstract T25 is one of the 4 maize transformation events from which commercial lines have so far been authorized in Europe. It was created by polyethylene glycol-mediated transformation using a construct bearing one copy of the synthetic pat gene associated with both promoter and terminator of the 35S ribosomal gene from cauliflower mosaic virus. In this article, we report the sequencing of the whole T25 insert and the characterization of its integration site by using a genome walking strategy. Our results confirmed that one intact copy of the initial construct had been integrated in the plant genome. They also revealed, at the 5′ junction of the insert, the presence of a second truncated 35S promoter, probably resulting from rearrangements which may have occurred before or during integration of the plasmid DNA. The analysis of the junction fragments showed that the integration site of the insert presented high homologies with the Huck retrotransposon family. By using one primer annealing in the maize genome and the other in the 5′ end of the integrat ed DNA, we developed a reliable event-specific detection system for T25 maize. To provide means to comply with the European regulation, a real-time PCR test was designed for specific quantitation of T25 event by using Taqman® chemistry.

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Determination of Butaperazine in Biological Fluids by Gas Chromatography Using Nitrogen Specific Detection System

Journal of Chromatographic Science ◽

10.1093/chromsci/17.12.666 ◽

1979 ◽

Vol 17 (12) ◽

pp. 666-670 ◽

Cited By ~ 8

Author(s):

J. I. Javaid ◽

H. Dekirmenjian ◽

U. Liskevych ◽

J. M. Davis

Keyword(s):

Gas Chromatography ◽

Detection System ◽

Biological Fluids ◽

Specific Detection

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High Resolution Screening of Plant Natural Product Extracts for Estrogen Receptor a and f3 Binding Activity Using an Online HPLC-MS Biochemical Detection System

CrossRef Listing of Deleted DOIs ◽

10.1177/108705710100600503 ◽

2001 ◽

Vol 6 (5) ◽

pp. 291-303 ◽

Cited By ~ 3

Author(s):

Uwe Schobel ◽

Michel Frenay ◽

Danny A. Van Elswijk ◽

Joanne M. McAndrews ◽

Kelly R. Long ◽

...

Keyword(s):

Estrogen Receptor ◽

High Resolution ◽

Natural Product ◽

High Speed ◽

Detection System ◽

Binding Activity ◽

Biologically Active ◽

Fluorescence Detector ◽

Active Compounds ◽

Activation Assay

A new screening technology that combines biochemical analysis with the resolution power of high-performance liquid chromatography (HPLC), referred to here as high-resolution screening (HRS) technique, is described. The capability of the HRS technology to analyze biologically active compounds in complex mixtures is demonstrated by screening a plant natural product extract library for estrogen receptor (ER) a and fi binding activity. The simultaneous structure elucidation of biologically active components in crude extracts was achieved by operating the HRS system in combination with mass spectrometry (MS). In contrast to conventional microtiter-type bioassays, the interactions of the extracts with the ER and the employed label, coumestrol, proceeded at high speed in a closed, continuous-flow reaction detection system, which was coupled directly to the outlet of a HPLC separation column. The reaction products of this homogeneous fluorescence enhancement-type assay were detected online using a flow-through fluorescence detector. Primary screening of the extract library was performed in the fast-flow injection analysis mode (FlowScreening) wherein the chromatographic separation system was bypassed. The library was screened at high speed, using two assay lines in parallel. A total of 98% of the identified hits were confirmed in a traditional 96-well microplate-based fluorescence polarization assay, indicating the reliability of the FlowScreening process. Active extracts were reassayed in a transcriptional activation assay in order to assess the functional activity of the bioactive extracts. Only functional active extracts were processed in the more time-consuming HRS mode, which was operated in combination with MS. Information on the number of active compounds, their retention times, the molecular masses, and the MS/MS-fingerprints as a function of their biological activity was obtained from 50% of the functional active extracts in real time. This dramatically enhances the speed of biologically active compound characterization in natural product extracts compared to traditional fractionation approaches.

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Rapid and Specific Measurements of Superoxide Using Fluorescence Spectroscopy

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057112468765 ◽

2012 ◽

Vol 18 (4) ◽

pp. 498-503 ◽

Cited By ~ 67

Author(s):

Rafal R. Nazarewicz ◽

Alfiya Bikineyeva ◽

Sergey I. Dikalov

Keyword(s):

Superoxide Dismutase ◽

Fluorescence Spectroscopy ◽

High Performance ◽

Hplc Analysis ◽

Cultured Cells ◽

Oxidation Products ◽

Specific Detection ◽

Intact Cells ◽

Specific Product ◽

Plate Reader

Superoxide plays a key role in many pathological processes; however, detection of superoxide by one of the most common methods using dihydroethidium (DHE) may be unspecific because of overlapping fluorescence of the superoxide-specific product, 2-OH-ethidium (2OH-E), and the unspecific oxidation product, ethidium. Here, we show a new optimized fluorescence spectroscopy protocol that allows rapid and specific detection of superoxide in cell-free systems and intact cells using DHE. We defined new optimized fluorescent settings to measure the superoxide-specific product and minimize the interference of unspecific DHE oxidation products. Using this protocol, we studied real-time superoxide production by xanthine oxidase– and menadione-treated cultured cells. Specificity of the plate reader–based superoxide measurements was confirmed by the inhibition of fluorescence with superoxide dismutase and high-performance liquid chromatography (HPLC) analysis. We show that limitations of the HPLC-based analysis can be overcome by the optimized fluorescence spectroscopy.

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Specific Detection of Legionella in Samples from Patients with Community-Acquired Pneumonia by PCR and a Colorimetric Detection System (Reverse Dot Blot)

Legionella ◽

10.1128/9781555815660.ch13 ◽

2014 ◽

pp. 51-52

Author(s):

Jörn Kircher ◽

Alexander Kirchhoff ◽

Arndt Rolfs

Keyword(s):

Detection System ◽

Colorimetric Detection ◽

Community Acquired Pneumonia ◽

Dot Blot ◽

Specific Detection ◽

Reverse Dot Blot

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Design of Universal Detection System for Automobile Combination Dashboard

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.274.350 ◽

2013 ◽

Vol 274 ◽

pp. 350-354

Author(s):

Hui Juan Yuan ◽

Wei Liu ◽

Wei Hu ◽

Yong Quan Sun

Keyword(s):

Detection System ◽

Electronic Control Unit ◽

Control Unit ◽

Motor Vehicles ◽

Maximum Relative Error ◽

Electronic Control ◽

Specific Detection ◽

Detection Systems ◽

Automatic Detection System ◽

Universal Detection

The detection of automotive combination dashboard is usually implemented by adopting the specific detection system matched with models, and automotive combination dashboard of different models requires different detection systems. A universal automatic detection system is designed in this paper, which can detect different kinds and models of automotive combination dashboard. First, standard detection signal generated by the electronic control unit is added to every corresponding position of dashboard, second, cameras film the corresponding images of dashboard in the standard signal excitation, last, the image is sent to the host computer for image processing to get output data of dashboard. This system detects results of automotive combination dashboard with the maximum relative error less than 2.5%, which meets the specified accuracy requirements in the automotive industrial standard QC/T727-2007 “Instruments for Motor Vehicles and Motorcycles”

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