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ABSTRACT: Comment by Tsikas et al. (Analyst, 2011, DOI: ) on the preliminary work by Shen and colleagues (Analyst, 2010, 135, 302) describing the use of ESI-MS/MS for the detection of methylpiperazinobenzenediamine, as a probe for NO in tissue samples, have been addressed in this communication. The article concludes that the points raised by Tsikas represent a logical progression to the research reported and indicate the way forward in realising appropriate analytical methodology.
The Analyst 10/2010; · 4.23 Impact Factor
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ABSTRACT: This review focuses on the development and use of microfluidic devices within a clinical setting. The underlying theoretical background of microfluidics is briefly elucidated. The materials and techniques used to fabricate the devices and their applicability to the clinical environment are described. The current research in this area is appraised and projections for future applications are discussed. Copyright © 2010 Society of Chemical Industry
Journal of Chemical Technology & Biotechnology 08/2010; 86(1):10 - 17. · 2.17 Impact Factor
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ABSTRACT: A novel method to determine nitric oxide (NO) in biological tissue samples with minimal interference from the cellular detritus is described. Methylpiperazinobenzenediamine, consisting of an o-phenylenediamine and a methyl piperazine group, was chosen as a probe for the detection of NO by mass spectrometry (MS) in biological tissue samples. The o-phenylenediamine group reacts with NO to form a characteristic benzotriazole. The product was identified using electrospray ionization mass spectrometry (ESI-MS) and the method validated within the range of 95-1900 nM. NO levels associated with tissue biopsies (approximately 10 mg) from rat vasculature and intestine tissue biopsies have been successfully determined. The different rates of NO generated from tissue samples under hypoxic and normoxic conditions have been studied by this simple and sensitive method.
The Analyst 02/2010; 135(2):302-5. · 4.23 Impact Factor
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ABSTRACT: Sol-gel nanoprobes, also known as Photonic Explorer for Bioanalysis with Biologically Localised Embedding (PEBBLE), capable of performing in-vitro intracellular monitoring of reactive oxygen species have been developed using a modified form of 5(6)-carboxyfluorescein diacetate. A sol-gel matrix was selected for the design of the probes as it is photostable, optically transparent and chemically inert, and to minimise leaching of the dye from the porous matrix it was covalently immobilised to silica nanoparticles (15 nm). Using this approach, 0.1% of the dye was found to leach over a typical analysis time of 5 h and minimal photobleaching was observed. In addition, the nanoprobes were shown to respond to hydrogen peroxide, hydroxyl anions, nitric oxide, peroxynitrile and superoxide anions, obtaining limits of detection of 2.2, 1.1, 3.2, 1.1 and 1.1 nM respectively. The nanoprobes were subsequently introduced into bovine oviducts using a lipid transfection reagent (Escort IV) and fluorescence was observed.
The Analyst 02/2008; 133(1):71-5. · 4.23 Impact Factor
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ABSTRACT: Probes Encapsulated By Biologically Localised Embedding (PEBBLEs) are optical sensors with nanometer dimensions fabricated by microemulsion polymerisation. The most beneficial characteristic of these sensors is the protection offered by the sensor matrix which decreases interaction between the fluorophore and the cell. These sensors have been introduced to the cell by a number of methods; however this paper discusses the development of a generic method to facilitate inclusion of this type of sensor in the cell by a simple incubation step. This was achieved by covalent linkage of a synthetic Cell Penetrating Peptide (CPP) based on the Human Immuno-deficiency Virus (HIV) -1 Tat, to the external sensor matrix. Calcium sensors were used to demonstrate this approach to incorporate the sensors within the cell. Characterisation revealed the calcium sensors were approximately 30 +/- 7 nm in diameter with a slightly negative zeta potential. The sensors demonstrated a linear range of 0-50 microM with negligible interference from a range of cellular ions and protein. Leaching of entrapped dyes from the calcium sensors was determined as 3% in a 24 h period, while photobleaching of the entrapped dye was minimal over a 40 min period. The sensors ability to cross the cell membrane using the covalently attached synthetic Tat peptide is demonstrated. Cellular inclusion of the sensors occurred within a 30 min incubation period.
The Analyst 03/2005; 130(2):163-70. · 4.23 Impact Factor
