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1.
Peptides ; 28(2): 269-80, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17194505

ABSTRACT

We have utilized a rat model of peripheral artery disease (PAD) to examine whether the known angiogenic activity of the Y(2) receptor would translate into a meaningful increase in collateral blood flow. The maximal increase in collateral blood flow capacity of approximately 60% (p<0.001) was obtained with a 10microg/kgday (IA infusion, 14 days) of either PYY or PYY(3-36) and did not differ from that obtained with a maximally angiogenic dose of VEGF(165). Pharmacodynamic modeling based upon single dose pharmacokinetic plasma profiles of both agonists suggests that E(max) is reached when the Y(2) receptor is occupied by >or=50%. Furthermore, for PYY(3-36), occupancy of the Y(2) receptor is sufficient to promote a significant benefit in collateral blood flow.


Subject(s)
Blood Circulation/physiology , Models, Biological , Peripheral Vascular Diseases/metabolism , Receptors, Neuropeptide Y/physiology , Animals , Base Sequence , DNA Primers , Female , Humans , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction
2.
Electrophoresis ; 24(9): 1375-82, 2003 May.
Article in English | MEDLINE | ID: mdl-12731022

ABSTRACT

The separation and detection of complexes of aptamers and protein targets by capillary electrophoresis (CE) with laser-induced fluorescence was examined. Aptamer-thrombin and aptamer-immunoglobulin E (IgE) were used as model systems. Phosphate, 3-(N-morpholino)propanesulfonic acid with phosphate, and tris(hydroxyamino)methane-glycine-potassium (TGK) buffer at pH 8.4 were tested as electrophoresis media. Buffer had a large effect with TGK providing the most stable complexes for both protein-aptamer complexes. Conditions that suppressed electroosmotic flow, such as addition of hydroxypropylmethylcellulose to the media or modification of the capillary inner wall with polyacrylamide, were found to prevent detection of complexes. The effect of separation time and electric field were evaluated by monitoring complexes with electric field varied from 150-2850 V/cm and effective column lengths of 3.5 and 7.0 cm. As expected, shorter times on the column greatly increased peak heights for the complexes due to a combination of less dilution by diffusion and less dissociation on the column. High fields were found to have a detrimental effect on detection of complexes. It is concluded that the best conditions for detection of noncovalent complexes involve use of the minimal column length and electric field necessary to achieve separation. The results will be of interest in developing affinity probe CE assays wherein aptamers are used as affinity ligands.


Subject(s)
Chromatography, Affinity/methods , Electrophoresis, Capillary/methods , Oligonucleotides/chemistry , Proteins/isolation & purification , Animals , Buffers , DNA, Single-Stranded , Humans , Immunoglobulin E/isolation & purification , Ligands , Rheology , Static Electricity , Thrombin/isolation & purification , Time Factors
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