For many disease states, positive outcomes are directly linked to early diagnosis, where therapeutic intervention would be most effective. Recently, trends in disease diagnosis have focused on the development of label-free sensing techniques that are sensitive to low analyte concentrations found in the physiological environment. Surface-enhanced Raman spectroscopy (SERS) is a powerful vibrational spectroscopy that allows for label-free, highly sensitive, and selective detection of analytes through the amplification of localized electric fields on the surface of a plasmonic material when excited with monochromatic light. This results in enhancement of the Raman scattering signal, which allows for the detection of low concentration analytes, giving rise to the use of SERS as a diagnostic tool for disease. Here, we present a review of recent developments in the field of in vivo and in vitro SERS biosensing for a range of disease states including neurological disease, diabetes, cardiovascular disease, cancer, and viral disease.

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Cortisol is an important steroid hormone in human physiology. Variations or abnormalities in the physiological cortisol levels control acute and chronic stress response, as well as contribute to diseases and syndromes including Addison's disease and Cushing syndrome. The ability to monitor cortisol levels in the physiological range is key in diagnosis and monitoring of these conditions, where current methodology for determination of cortisol levels relies on instrumentation that requires extensive sample preparation, long run times, and is destructive to the sample. Raman spectroscopy provides rapid sample analysis with relatively simple instrumentation; however, Raman spectroscopy is an inherently weak technique. To provide an enhanced Raman signal, we use surface enhanced Raman spectroscopy (SERS) which utilizes oscillating electric fields of metal nanoparticles, enhancing the overall electric field and therefore resulting in an enhanced signal. We demonstrate SERS-based detection of cortisol in the physiologically relevant range using colloidal silver nanoparticles in ethanolic solutions and bovine serum albumin. The SERS spectra obtained in an ethanol matrix demonstrate a sigmoidal concentration response over the physiologically relevant concentration range, with a limit of detection established at 177 nM. Analysis of cortisol solutions in a complex matrix (bovine serum albumin in phosphate buffered saline) is also demonstrated through the use of principal components analysis, a multivariate technique, which shows the separation of cortisol in a linear fashion with respect to cortisol concentration.

The carbonate radical anion CO(3)(•-) is a potent reactive oxygen species (ROS) produced in vivo through enzymatic one-electron oxidation of bicarbonate or, mostly, via the reaction of CO(2) with peroxynitrite. Due to the vitally essential role of the carbon dioxide/bicarbonate buffer system in regulation of physiological pH, CO(3)(•-) is arguably one of the most important ROS in biological systems. So far, the studies of reactions of CO(3)(•-) with DNA have been focused on the pathways initiated by oxidation of guanines in DNA. In this study, low-molecular products of attack of CO(3)(•-) on the sugar-phosphate backbone in vitro were analyzed by reversed phase HPLC. The selectivity of damage in double-stranded DNA (dsDNA) was found to follow the same pattern C4' > C1' > C5' for both CO(3)(•-) and the hydroxyl radical, though the relative contribution of the C1' damage induced by CO(3)(•-) is substantially higher. In single-stranded DNA (ssDNA) oxidation at C1' by CO3(•-) prevails over all other sugar damages. An approximately 2000-fold preference for 8-oxoguanine (8oxoG) formation over sugar damage found in our study identifies CO(3)(•-) primarily as a one-electron oxidant with fairly low reactivity toward the sugar-phosphate backbone.

A novel analytical high-performance liquid chromatography (HPLC)-based method of quantification of the yields of C4'-oxidized abasic sites, 1, in oxidatively damaged DNA has been elaborated. This new approach is based on efficient conversion of 1 into N-substituted 5-methylene-Δ(3)-pyrrolin-2-ones, 2, upon treatment of damaged DNA with primary amines in neutral or slightly acidic solutions with subsequent quantification of 2 by HPLC. The absolute and relative radiation-chemical yields of 1 in irradiated DNA solutions were re-evaluated using this method. The yields were compared with those of other 2-deoxyribose degradation products including 5-methylene-2(5H)-furanone, malondialdehyde, and furfural resulting from the C1', C4' and C5'-oxidations, respectively. The yield of free base release (FBR) determined in the same systems was employed as an internal measure of the total oxidative damage to the 2-deoxyribose moiety. Application of this technique identifies 1 as the most abundant sugar lesion in double-stranded (ds) DNA irradiated under air in solution (36% FBR). In single-stranded (ss) DNA this product is second by abundance (33% FBR) after 2-deoxyribonolactones (C1'-oxidation; 43% FBR). The production of nucleoside-5'-aldehydes (C5'-oxidation; 14% and 5% FBR in dsDNA and ssDNA, respectively) is in the third place. Taken together with the parallel reaction channel that converts C4'-radicals into malondialdehyde and 3'-phosphoglycolates, our results identify the C4'-oxidation as a prevalent pathway of oxidative damage to the sugar-phosphate backbone (50% or more of all 2-deoxyribose damages) in indirectly damaged DNA.