Analysis of intracellular enzyme activity by surface enhanced Raman scattering.

Dysfunctional intracellular enzymatic activity is believed to be an underlying cause of a myriad of diseases. We present the first use of surface enhanced Raman scattering (SERS) as a detection technique capable of reporting intracellular activity of a specific enzyme. Careful choice of reagents allowed the preparation of high resolution cellular activity maps highlighting the specific conversion of the commonly used ELISA reagent 5-bromo-4-chloro-3-indolyl ß-D-galactopyranoside (X-Gal), by wild type ß-galactosidase enzymes. Further, through co-addition of X-Gal substrate and inhibitors we were able to demonstrate that intracellular substrate conversion occurred predominantly through an enzymatically specific pathway. The data presented therefore supports the application of SERS probes as sensitive, specific sensors of biochemical activity and demonstrates the use of SERS probes for the first time as beacons capable of high resolution subcellular localisation of native enzymes.

The effect of business improvement districts on the incidence of violent crimes.

OBJECTIVE: To examine whether business improvement districts (BID) contributed to greater than expected declines in the incidence of violent crimes in affected neighbourhoods. METHOD: A Bayesian hierarchical model was used to assess the changes in the incidence of violent crimes between 1994 and 2005 and the implementation of 30 BID in Los Angeles neighbourhoods. RESULTS: The implementation of BID was associated with a 12% reduction in the incidence of robbery (95% posterior probability interval -2 to 24) and an 8% reduction in the total incidence of violent crimes (95% posterior probability interval -5 to 21). The strength of the effect of BID on robbery crimes varied by location. CONCLUSION: These findings indicate that the implementation of BID can reduce the incidence of violent crimes likely to result in injury to individuals. The findings also indicate that the establishment of a BID by itself is not a panacea, and highlight the importance of targeting BID efforts to crime prevention interventions that reduce violence exposure associated with criminal behaviours.

The effect of light rail transit on body mass index and physical activity.

BACKGROUND: The built environment can constrain or facilitate physical activity. Most studies of the health consequences of the built environment face problems of selection bias associated with confounding effects of residential choice and transportation decisions. PURPOSE: To examine the cross-sectional associations between objective and perceived measures of the built environment; BMI; obesity (BMI>30 kg/m(2)); and meeting weekly recommended physical activity (RPA) levels through walking and vigorous exercise. To assess the effect of using light rail transit (LRT) system on BMI, obesity, and weekly RPA levels. METHODS: Data were collected on individuals before (July 2006-February 2007) and after (March 2008-July 2008) completion of an LRT system in Charlotte NC. BMI, obesity, and physical activity levels were calculated for a comparison of these factors pre- and post-LRT construction. A propensity score weighting approach adjusted for differences in baseline characteristics among LRT and non-LRT users. Data were analyzed in 2009. RESULTS: More-positive perceptions of one's neighborhood at baseline were associated with a -0.36 (p<0.05) lower BMI; 15% lower odds (95% CI=0.77, 0.94) of obesity; 9% higher odds (95% CI=0.99, 1.20) of meeting weekly RPA through walking; and 11% higher odds (95% CI=1.01, 1.22) of meeting RPA levels of vigorous exercise. The use of LRT to commute to work was associated with an average -1.18 reduction in BMI (p<0.05) and an 81% reduced odds (95% CI=0.04, 0.92) of becoming obese over time. CONCLUSIONS: The results of this study suggest that improving neighborhood environments and increasing the public's use of LRT systems could provide improvements in health outcomes for millions of individuals.

Dynamic Imaging Analysis of SERS-Active Nanoparticle Clusters in Suspension.

A novel wide-field approach for the real-time Surface Enhanced Raman Scattering (SERS) imaging of multiple silver nanoparticle clusters suspended in solution is described. This method enables direct correlation of the SERS activity of a single nanoparticle aggregate and its size through measurement of the cluster diffusion coefficient and can also be performed in a high-throughput basis. As a first demonstration, we investigate the salt-induced aggregation of silver nanoparticles in the presence of a reporter tag molecule, which has a high affinity for the nanoparticle surface. In addition to tracking individual particles, direct comparison of Rayleigh and SERS videos of the same colloid solution enabled measurement of the fraction of individual clusters that are SERS active and the dependence of this value on the relative concentration of the tag molecule. Furthermore, given the ability to also rapidly profile any nonuniformity in particle size distributions, we expect this approach will not only provide a new tool for the fundamental understanding of SERS but also significantly contribute to the development of an array of emerging nanoparticle-enhanced biomolecule and imaging detection platforms.

Imaging inflammation in real time--future of nanoparticles.

The detection of subclinical early inflammation in autoimmune diseases is an important but currently technically demanding approach to direct initial diagnosis and subsequent choice of therapy. Recent advances in imaging using NP provides the potential to detect cellular recruitment, vascular activation or leakage at a subclinically stage of disease and may provide predictive "biomarkers" of future pathogenesis. The NP used are either untargeted and taken up by phagocytic cells, or are linked to a ligand, targeting localisation to the site of inflammation. Techniques, varying from MRI and fluorescence to Raman spectroscopy are being employed. In this short review, we summarise many of the recent developments in the field of NP imaging related to inflammation.

Rapid cell mapping using nanoparticles and SERRS.

Bone marrow-derived immune cells (macrophages) treated with gold and silver nanoparticles before fixation and dye staining have been analysed by multiple wavelength line scanning surface enhanced resonance Raman scattering (SERRS) mapping. The method yields high selectivity and sensitivity within short analysis times, identifying nanoparticle aggregates in secondary lysosomes. Using routine cell stains, the output from fluorescence, Raman and SERRS is quantified at four wavelengths of excitation, demonstrating the potential at longer biologically compatible wavelengths of using nanoparticles with cell stains for superior cell mapping.

In situ detection of pterins by SERS.

Surface enhanced Raman scattering (SERS) has been used to detect specific pterin molecules at sub-nanomolar concentrations. SERS is fast becoming a widely used technique for the sensitive and specific detection of multiple analytes. The information-rich and concentration-dependent spectra obtained from SERS make the technique ideally placed for high speed, low cost analysis of almost any analyte. Further, to show the feasibility of SERS in the detection of biologically relevant targets, a synthetic pterin analogue of the naturally occurring pterin cofactor, tetrahydrobiopterin, has been detected at a series of concentrations and the method used for the successful detection of the synthetic pterin in mouse serum. In this analysis, spectroscopic collection was optimized for water-based pteridine derivatives using two visible wavelengths of excitation (514.5 and 632.8 nm) and differing mesoscopic metal nanoparticles allowing the limits of detection to be calculated.

Dip-pen nanolithography and SERRS as synergic techniques.

We demonstrate the powerful combination of dip-pen nanolithography (DPN) performed on non-flat plasmonic gold surfaces and subsequent detection by surface enhanced resonance Raman scattering (SERRS).

Immunoassay for P38 MAPK using surface enhanced resonance Raman spectroscopy (SERRS).

A micro-bead sandwich assay for P38 mitogen-activated protein kinase using surface enhanced resonance Raman spectroscopy (SERRS) detection is reported. Monoclonal capture antibodies were immobilised on a solid phase of magnetic micro-beads with secondary detection using a rhodamine-labelled antibody. Quantitative SERRS detection of the secondary antibody was possible with a limit of detection of 9.5 x 10(-12) mol dm(-3). The sandwich assay was quantitative and sensitive to 6 ng ml(-1). The mechanism of the SERRS detection in the immunoassay was investigated. The addition of SERRS aggregating agents causes the dissociation of the immuno-complex from the magnetic beads. Scanning electron microscopy images indicate that the colloidal suspension rather than adsorbed silver nanoparticles on the beads provide the SERRS signals, that the aggregate size is partially controlled and that there is some inhomogeneity in the distribution of organic matter on the nanoscale.

Surface-enhanced Raman scattering spectroscopy as a sensitive and selective technique for the detection of folic acid in water and human serum.

Surface-enhanced Raman scattering (SERS) is shown to give linear and sensitive concentration-dependent detection of folic acid using silver nanoparticles created via ethylene-diaminetetraacetic acid (EDTA) reduction. Optical detection by SERS overcomes the primary limitation of photodissociation encountered during the application of other shorter wavelength ultraviolet (UV)/near-UV techniques such as fluorescence based microscopy. The SERS approach in water-based samples was demonstrated and optimized using several longer wavelengths of excitation (514.5, 632.8, and 785 nm). Excitation in the green (514.5 nm) was found to achieve the best balance between photodissociation and SERS efficiency. Linear concentration dependence was observed in the range of 0.018 to 1 microM. The importance of folic acid in a clinical setting and the potential applications of this technique in a biological environment are highlighted. We demonstrate the potential to transfer this technique to real biological samples by the detection of folic acid in human serum samples by SERS.

Squaraines as unique reporters for SERRS multiplexing.

Modified anilino squaraine dyes provide unique SERRS spectra that can be identified at low concentrations within any mixture of current reporters using longer, biologically compatible wavelengths of excitation.

Estimating the effects of light rail transit on health care costs.

In recent years, there has been a proliferation of research on the effects of the built environment, including mass transit systems, on health-related outcomes. While there is general agreement that the built environment affects travel choices and physical activity, it remains unclear how much of a public health benefit (in dollars) can be derived from land use policies that support walking, biking, and transit. In the present study, we develop a model to assess the potential cost savings in public health that will be realized from the investment in a new light rail transit system in Charlotte, NC. Relying on estimates of future riders, area obesity rates, and the effects of public transit on physical activity (daily walking to and from the transit stations), we simulated the potential yearly public health cost savings associated with this infrastructure investment. Our results indicate that investing in light rail is associated with a 9-year cumulative public health cost savings of dollars 12.6 million. While these results suggest that there is a sizable public health benefit associated with the adoption of light rail, they also indicate that the effects are relatively small compared to the costs associated with constructing and operating such systems. These findings suggest that planning efforts that focus solely on the health impact of modifications in the built environment are likely to overstate the economic benefits. Public health benefits should be considered along with broader environmental health benefits.

8-hydroxyquinolinyl azo dyes: a class of surface-enhanced resonance Raman scattering-based probes for ultrasensitive monitoring of enzymatic activity.

A series of surface-enhanced resonance Raman scattering (SERRS) based probes for the detection of lipase activity are reported. A number of novel SERRS-active 8-hydroxylquinolinyl azo dyes have been prepared and via synthetic esterification or subsequent enzymatic hydrolysis at the 8-hydroxyl position the SERRS signal can be "switched" on or off. In the first instance, the technique has been demonstrated for the successful detection of lipase from Pseudomonas cepacia, and these new compounds offer a limit of detection of 0.2 ng mL-1 enzyme, up to a 100-fold lower limit than observed for benzotriazolyl dyes used in previous studies. The chemical synthesis is straightforward and allows for facile introduction of a wide range of different masking groups, using commonly known synthetic methodologies. The potential for multiplexing analysis of enzyme activity using this technology is presented within.

Quantitative enhanced Raman scattering of labeled DNA from gold and silver nanoparticles.

Surface-enhanced resonance Raman scattering (SERRS) from silver nanoparticles using 514.5-nm excitation has been shown to offer huge potential for applications in highly sensitive multiplexed DNA assays. If the technique is to be applied to real biological samples and integrated with other methods, then the use of gold nanoparticles and longer wavelengths of excitation are desirable. The data presented here demonstrate that dye-labeled oligonucleotide sequences can be directly detected by SERRS using gold nanoparticles in a quantitative manner for the first time. The performance of gold and silver nanoparticles as SERRS substrates was assessed using 514.5-, 632.8-, and 785-nm excitation and a range of 13 commercially available dye-labeled oligonucleotides. The quantitative response allowed the limit of detection to be determined for each case and demonstrates that the technique is highly effective, sensitive, and versatile. The possibility of excitation at multiple wavelengths further enhances the multiplexing potential of the technique. The importance of effectively combining the optical properties of the nanoparticle and the dye label is demonstrated. For example, at 632.8-nm excitation, the dye BODIPY TR-X and gold nanoparticles make a strong SERRS combination with very little background fluorescence. This study allows the choice of nanoparticle and dye label for particular experimental setups, and significantly expands the applicability of enhanced Raman scattering for use in many disciplines.

Highly sensitive detection of dye-labelled DNA using nanostructured gold surfaces.

Careful control of surface chemistry results in strong surface enhanced resonance Raman scattering from dye-labelled oligonucleotides assembled on nanostructured gold surfaces, releasing their potential as reliable enhancing surfaces.