Patterned Protein Microarrays for Bacterial Detection

Abstract Patterned microarrays of antibodies were fabricated and tested for their ability to bind targeted bacteria. These arrays were used in a series of bacterial immunoassays to detect E. coli 0157:H7 and Renibacterium Salmoninarum (RS). Microarrays were fabricated using microcontact printing (µCP) and characterized using scanning probe microscopy (SPM). The high-resolution SPM imaging showed that targeted bacteria had a higher binding selectivity to complementary antibody patterns than to unfunctionalized regions of the substrate. Additional studies indicated a significant reduction in binding of bacteria when the microarrays were exposed to non-specific bacteria. These studies demonstrate how protein microarrays could be developed into useful platforms for sensing microorganisms.

New Coordinative Compounds with 4-(4’-pyridyl)pyridinium Disubstituted Monoylides

The complexes with manganese(II), iron(II), cobalt(II), nickel(II) and copper(II) of 2-(4, 4’-bipyridin-1-ium-1-yl)-1-(4-bromophenylamino)-3-(4-methoxyphenyl)-3-oxo-1-thioxopropan-2-ide (ylide, Y) were synthesized and characterized. The obtained compounds with 1 : 2 metal/ligand ratios have been characterized by FTIR, UV Vis spectroscopy, ESI MS spectrometry, molecular conductance, magnetic measurements and thermal analysis. The ylide ligand forms chelates with metallic (II) ions through their amide nitrogen and oxygen atoms

Hierarchical clustering of MS peak profiles of ISE6 cells treated with LGTV, UV-LGTV, and mock.

<p>M1-5, mock1-5 samples; LGTV1-5, LGTV-infected samples 1–5, and UV-LGTV1-5, UV-LGTV samples 1–5. Vertical rows depict n = 5 biological replicates. Horizontal rows correspond to significant MS peaks of peptides/proteins at 36 hours post infection/treatment. Clustering analysis shows common patterns of protein expression profiles shared between the three treatment groups. The red-green color scale denotes the Z score fold change with red representing a Z score of -2 and green denoting a Z score of 2 [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004180#pntd.0004180.ref041" target="_blank">41</a>].</p

Citrate cycle showing ISE6 proteins that exhibited increased/decreased expression following treatment to LGTV and UV-LGTV.

<p>The enzymes are indicated with KEGG abbreviated nomenclature and the corresponding substrates are shown in circles. * denotes proteins identified in this study. Dotted lines denote indirect involvement with production. The increased expression of malate dehydrogenase (MDH2) is unique to LGTV-treated cells while increase in the expression of acetyl-CoA acetyltransferase 1 (ACAT1), delta-1-pyrroline-5-carboxylate dehydrogenase (ALDH4A1), glutamate dehydrogenase (GLUD1), and fumarylacetoacetase (FAH) is common to cell samples following LGTV infection and UV-LGTV treatment. Decreased expression of citrate synthase (CS) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was observed in LGTV-treated cells and decreased expression of fumarate hydratase (FH), aldolase A/B/C fructose-bisphosphate (ALDOA/B/C), and aldehyde dehydrogenase 2/1B1/3A2 family protein (ALDH2/1B1/3A2) was observed in both LGTV-infected and UV-LGTV—treated cells. ATP citrate lyase (ACLY), aconitase (ACO), isocitrate dehydrogenase 2/3a (IDH2/3a), oxoglutarate/alpha-ketoglutarate dehydrogenase complex (OGDH/DLST), succinyl-CoA synthetase alpha/beta subunit (LSC1/2), succinate dehydrogenase flavoprotein subunit (SDHA), pyruvate kinase (PK), enolase 1/2/3 (ENO1/2/3), and aldehyde dehydrogenase 7A1 family protein (ALDH7A1).</p