A new issue of this journal has just been published. To see abstracts of the papers it contains (with links through to the full papers) click here:
Selected papers from the latest issue:
PCR-Ready Human DNA Extraction from Urine Samples Using Magnetic Nanoparticles
Publication year: 2011
Source: Journal of Chromatography B, Available online 6 December 2011
Zhi Shan, Zhongwu Zhou, Hui Chen, Zhiming Zhang, Yi Zhou, ...
Urine-derived human genomic DNA (gDNA) has wide application in a variety of disciplines including clinical medicine, sports, and forensic science. We describe a novel method for gDNA extraction from urine samples using carboxylated magnetic nanoparticles (CMNPs) as solid-phase adsorbents. Sedimentation associated with freezing of urine samples significantly reduces cell capture by CMNPs. However, the addition of 10 mM EDTA and subsequent pH modification (pH 6.0-7.1) can re-dissolve urine sediments. Purified gDNA ranged from around 0.1 kb to more than 23 kb. PCR using specific primers targetingK-ras,GAPDH,CYP3A4andGDF5amplified 100% of varying sized gene fragments, verifying the high quality of the isolated DNA. Successful PCR amplifications using DNA isolated from urine samples as small as 50 μl were demonstrated. Enrichment of urine cells and subsequent adsorption of DNA can be achieved with the same CMNPs, greatly simplifying extraction procedures. The CMNP gDNA extraction technique proved to be simple, rapid, sensitive and environmentally friendly, with application for routine laboratory use and potentially within automated urine extraction platforms.
Source: Journal of Chromatography B, Available online 6 December 2011
Zhi Shan, Zhongwu Zhou, Hui Chen, Zhiming Zhang, Yi Zhou, ...
Urine-derived human genomic DNA (gDNA) has wide application in a variety of disciplines including clinical medicine, sports, and forensic science. We describe a novel method for gDNA extraction from urine samples using carboxylated magnetic nanoparticles (CMNPs) as solid-phase adsorbents. Sedimentation associated with freezing of urine samples significantly reduces cell capture by CMNPs. However, the addition of 10 mM EDTA and subsequent pH modification (pH 6.0-7.1) can re-dissolve urine sediments. Purified gDNA ranged from around 0.1 kb to more than 23 kb. PCR using specific primers targetingK-ras,GAPDH,CYP3A4andGDF5amplified 100% of varying sized gene fragments, verifying the high quality of the isolated DNA. Successful PCR amplifications using DNA isolated from urine samples as small as 50 μl were demonstrated. Enrichment of urine cells and subsequent adsorption of DNA can be achieved with the same CMNPs, greatly simplifying extraction procedures. The CMNP gDNA extraction technique proved to be simple, rapid, sensitive and environmentally friendly, with application for routine laboratory use and potentially within automated urine extraction platforms.
Highlights
► A method for urine DNA extraction using carboxylated magnetic nanoparticles was developed. ► The addition of 10 mM EDTA and pH modification (pH 6.0-7.1) can re-dissolve urine sediments. ► Purified DNA ranged from around 0.1 kb to more than 23 kb. ► DNA quality was validated by its yield, molecular weight, and the ability to serve as PCR templates. ► The developed method proved to be simple, rapid, sensitive and environmentally friendly.Competitive binding between 4,4’-diphenylmethane-bis(methyl) carbamate and RAGE ligand MG-H1 on human umbilical vein endothelial cell by cell membrane chromatography
Publication year: 2011
Source: Journal of Chromatography B, Available online 6 December 2011
Liang Feng, You-hua Xu, Shan-shan Wang, Wai Au-yeung, Zhao-guang Zheng, ...
The compound 4,4’-diphenylmethane-bis(methyl) carbamate (CM1) has a protective activity on AGEs-induced endothelial dysfunction on human umbilical vein endothelial cell (HUVEC) in our previous study. It suggested that CM1 which may act as a competitive antagonist to the blockade of AGEs to receptor of AGEs (RAGE) and attenuate the HUVEC damage. In order to testify that hypothesis, the cell membrane chromatography (CMC) combined with high performance liquid chromatography (HPLC) was developed for analyzing the competitive binding properties on RAGE of HUVEC between CM1 and MG-H1, the agonist of RAGE. The results from saturation binding of CM1 and MG-H1 on cells demonstrated that dissociation equilibrium constants (Kd) of CM1 and MG-H1 were 3.653 nM and 4.12 nM, respectively; While maximum binding capacity (Bmax) of CM1 and MG-H1 were 30.08 and 18.72 fmol/mg protein, respectively. In competition experiments, IC50of CM1 with pre-incubation 10 M and 10 M MG-H1 were 1.37 × 10 M and 4.56 × 10 M, respectively. The present findings indicated that CM1 conjugated competitively to cells with RAGE ligand MG-H1. The primary study illustrated that CMC combined with HPLC analysis method could be an alternative, rapid and efficient approach for the interaction of drug molecule and receptor, and that CM1 intervene the AGEs inducing HUVEC damage may via the competitively block the AGEs-RAGE path way.
Source: Journal of Chromatography B, Available online 6 December 2011
Liang Feng, You-hua Xu, Shan-shan Wang, Wai Au-yeung, Zhao-guang Zheng, ...
The compound 4,4’-diphenylmethane-bis(methyl) carbamate (CM1) has a protective activity on AGEs-induced endothelial dysfunction on human umbilical vein endothelial cell (HUVEC) in our previous study. It suggested that CM1 which may act as a competitive antagonist to the blockade of AGEs to receptor of AGEs (RAGE) and attenuate the HUVEC damage. In order to testify that hypothesis, the cell membrane chromatography (CMC) combined with high performance liquid chromatography (HPLC) was developed for analyzing the competitive binding properties on RAGE of HUVEC between CM1 and MG-H1, the agonist of RAGE. The results from saturation binding of CM1 and MG-H1 on cells demonstrated that dissociation equilibrium constants (Kd) of CM1 and MG-H1 were 3.653 nM and 4.12 nM, respectively; While maximum binding capacity (Bmax) of CM1 and MG-H1 were 30.08 and 18.72 fmol/mg protein, respectively. In competition experiments, IC50of CM1 with pre-incubation 10 M and 10 M MG-H1 were 1.37 × 10 M and 4.56 × 10 M, respectively. The present findings indicated that CM1 conjugated competitively to cells with RAGE ligand MG-H1. The primary study illustrated that CMC combined with HPLC analysis method could be an alternative, rapid and efficient approach for the interaction of drug molecule and receptor, and that CM1 intervene the AGEs inducing HUVEC damage may via the competitively block the AGEs-RAGE path way.
Highlights
► CM1 competitively bind to RAGE with RAGE ligand MG-H1 ► A HPLC method was established for competition binding of CM1 with MG-H1 ► This method was an alternative way for competitive binding of drug to receptor ► This binding was performed on intact cells.The role of liquid chromatography-tandem mass spectrometry in the clinical laboratory
Publication year: 2011
Source: Journal of Chromatography B, Available online 6 December 2011
Johannes M.W. van den Ouweland, Ido P. Kema
Liquid chromatography coupled to mass spectrometry (LC-MS/MS) is increasingly used as a routine methodology in clinical laboratories for the analysis of low molecular weight molecules. The high specificity in combination with high sensitivity and multi-analyte potential makes it an attractive complementary method to traditional methodology used for routine applications. Its strength and weaknesses in this context will be discussed and examples of successful clinical applications will be given. For LC-MS/MS to truly fulfil its promise in clinical diagnosis, the prerequisite steps being sample pre-treatment, chromatographic separation and detection by selected reaction monitoring must become more integrated as they are in conventional clinical analyzers. The availability of ready-to-use reagents kits, eliminating efforts needed for method development and extensive validation, are likely to contribute to a wider acceptance of LC-MS/MS in clinical laboratories. Growing applicability of LC-MS/MS in the clinical laboratory field is expected from quantitative protein analysis.
Source: Journal of Chromatography B, Available online 6 December 2011
Johannes M.W. van den Ouweland, Ido P. Kema
Liquid chromatography coupled to mass spectrometry (LC-MS/MS) is increasingly used as a routine methodology in clinical laboratories for the analysis of low molecular weight molecules. The high specificity in combination with high sensitivity and multi-analyte potential makes it an attractive complementary method to traditional methodology used for routine applications. Its strength and weaknesses in this context will be discussed and examples of successful clinical applications will be given. For LC-MS/MS to truly fulfil its promise in clinical diagnosis, the prerequisite steps being sample pre-treatment, chromatographic separation and detection by selected reaction monitoring must become more integrated as they are in conventional clinical analyzers. The availability of ready-to-use reagents kits, eliminating efforts needed for method development and extensive validation, are likely to contribute to a wider acceptance of LC-MS/MS in clinical laboratories. Growing applicability of LC-MS/MS in the clinical laboratory field is expected from quantitative protein analysis.
Profiling and characterization of volatile secretions from the European stink bugGraphosoma lineatum(Heteroptera: Pentatomidae) by two-dimensional gas chromatography/time-of-flight mass spectrometry
Publication year: 2011
Source: Journal of Chromatography B, Available online 6 December 2011
Miloslav Šanda, Petr Žáček, Ludvík Streinz, Martin Dračínský, Bohumír Koutek
An efficient method combining the headspace solid-phase microextraction (HS-SPME) sampling procedure and comprehensive two-dimensional gas-chromatography/time-of-flight mass spectrometry (GCxGC/TOF-MS) was established to study the volatile secretion components of stink bugs (Heteroptera: Pentatomidae). The combined power of this approach is illustrated by the identification of fifty-seven compounds in the secretion of a European stink-bug representative,Graphosoma lineatum. (E)-4-oxohex-2-enal and (E)-dec-2-enal were found to be the major components in the adult bug secretions followed by lower amounts ofn-alkenal (C5–C12),n- alkenyl acetate (C5–C11),n-alkane (C11–C17) homologs, dienals and other compounds. More than thirty known compounds have been identified that had not been described before inG.lineatumadults. Of these compounds, (E)-4-oxohex-2-enal is of particular interest, since its isolation and identification, while calling some previous reports into question, clearly demonstrates a potential ability of our approach to yield artifact-free secretion profiles.
Source: Journal of Chromatography B, Available online 6 December 2011
Miloslav Šanda, Petr Žáček, Ludvík Streinz, Martin Dračínský, Bohumír Koutek
An efficient method combining the headspace solid-phase microextraction (HS-SPME) sampling procedure and comprehensive two-dimensional gas-chromatography/time-of-flight mass spectrometry (GCxGC/TOF-MS) was established to study the volatile secretion components of stink bugs (Heteroptera: Pentatomidae). The combined power of this approach is illustrated by the identification of fifty-seven compounds in the secretion of a European stink-bug representative,Graphosoma lineatum. (E)-4-oxohex-2-enal and (E)-dec-2-enal were found to be the major components in the adult bug secretions followed by lower amounts ofn-alkenal (C5–C12),n- alkenyl acetate (C5–C11),n-alkane (C11–C17) homologs, dienals and other compounds. More than thirty known compounds have been identified that had not been described before inG.lineatumadults. Of these compounds, (E)-4-oxohex-2-enal is of particular interest, since its isolation and identification, while calling some previous reports into question, clearly demonstrates a potential ability of our approach to yield artifact-free secretion profiles.
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