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The objective of this study was to evaluate the application of acute phase protein assays for C-reactive protein, haptoglobin, and serum amyloid A, as well as protein electrophoresis, in ferrets. Samples from 41 clinically normal and 52 clinically abnormal pet ferrets were submitted from veterinary clinics within the United States. Reference intervals were calculated for all analytes from the clinically normal group using the robust method. The clinically abnormal group demonstrated significant elevations in the concentrations of the following analytes ( P < 0.05): total protein, alpha 2 globulins, beta globulins, gamma globulins, serum amyloid A, and haptoglobin. The preliminary results of this research investigation support a potential role for these acute phase protein assays to be used as adjunct tests for monitoring inflammation and disease in ferrets. [ABSTRACT FROM AUTHOR]

Abstract: A series of bicyclic sulfonamides, 14–17, was prepared by thermal, intramolecular Diels–Alder cycloaddition. The ratio of exo:endo-diastereoisomers formed following this process was found to be dependent on the substitution pattern of the dienophile. Styryl substituted sulfonamides preferentially formed the corresponding exo-adducts, whereas vinyl substituted sulfonamides preferentially gave the endo-adducts. These adducts were treated under dissolved metal reduction conditions and it was found that compounds possessing an N-benzyl substituent underwent preferential debenzylation and resisted further reduction. The complementary N-isobutyl functionalised materials only underwent the double reduction when a phenyl substituent was present. [Copyright &y& Elsevier]

We compare immobilization methods for oligonucleotides on carboxylic acid surfaces with the goal of improving hybridization efficiency for single-stranded DNA (ssDNA) bioassays. When immobilized via 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), ssDNA (either modified with a terminal amine or not) specifically binds to the surface; without EDC, binding is minimal. EDC-activated probes can, however, bind covalently to the surface through nucleobase as well as terminal amino groups. Unmatched base pairs from the former are detected via melting curve analysis: the target begins to unwind at ∼40 °C below the double-strand melting temperature of ∼64 °C. To eliminate such backbone binding, we immobilized azide-functionalized DNA via click chemistry, resulting in hybridization efficiencies 5 times higher than with EDC. This improvement, and the room-temperature hybridization of the click chemistry process, make it an important alternative to EDC for reliable DNA assays with maximum specificity and sensitivity. [ABSTRACT FROM AUTHOR]

Silica nanoparticles (NPs) doped with far-red fluorescent cyanine dyes, Cy5 and FR670, were prepared using the microemulsion method. The effects of nucleation pathway on dye loading and NP morphology were investigated using UV–Vis spectroscopy and transmission electron microscopy for different combinations of three surfactants, Triton ® X-100, AOT and NP-5. Successful synthesis of monodispersed NPs with efficient dye loading was achieved using an intramicellar nucleation pathway combined with a negatively charged surfactant. For effective bioconjugation NPs were coated with a silica shell using either the microemulsion or the Stöber method. To maximise fluorescence intensity a series of NPs were doped with different amounts of dye ranging from 0.5 to 6% (w/w). At dye loadings of 0.5 and 0.25% (w/w) for Cy5 and FR670, respectively, NPs were 72 and 87 times more fluorescent than free dye labels. For Cy5 the change in fluorescence intensity with dye loading matched closely with a standard model for homo Förster resonance energy transfer (hFRET). A significant drop in fluorescence lifetime inside the NPs with increased dye loading was also observed and correlated with changes in energy transfer and quantum efficiency. Cy5 dye doped NPs were tested as biolabels in a fluorescence immunoassay to detect C-reactive protein, which is a recommended biomarker for cardiovascular disease. The NP label assay showed approximately an order of magnitude improvement in limit-of-detection when compared to a free dye assay. [ABSTRACT FROM AUTHOR]