Analysis of photothermal release of Oligonucleotides from hollow Gold nanospheres by surface enhanced raman scattering (SERS)

The photothermal release of single stranded DNA (ssDNA) from the surface of gold nanoparticles of different shapes and sizes is a promising mode of delivering DNA for gene-therapy applications. Here, we demonstrate the first targeted photothermal release of ssDNA from hollow gold nanospheres (HGNs) and analyse the release of the ssDNA using quantitative surface enhanced Raman scattering (SERS). The HGNs used demonstrate a tunable localized surface plasmon resonance (LSPR) frequency while maintaining size consistency, allowing for selective ssDNA release based on matching the excitation frequency to the plasmon resonance. It is shown that HGNs with resonances at 760 and HGN 670 nm release significant amounts of ssDNA when excited via 785 nm and 640 nm lasers respectively. When excited with a wavelength far from the LSPR of the particles, the ssDNA release is negligible. This is the first demonstration of SERS to analyze the amount of ssDNA photothermally released from the surface of HGNs. In contrast to traditional fluorescence measurements, this SERS based approach provides quantitatively robust data for analysis of ssDNA release and lays a strong foundation for future studies exploiting plasmonically induced ssDNA release

Characterisation of Estrogen Receptor Alpha (ERα) Expression in Breast Cancer Cells and Effect of Drug Treatment Using Targeted Nanoparticles and SERS

The detection and identification of estrogen receptor alpha (ERα), one of the main biomarkers in breast cancer, is crucial for the clinical diagnosis and therapy of the disease. Here, we use a non-destructive approach for detecting and localising ERα expression at the single cell level using surface enhanced Raman spectroscopy (SERS) combined with functionalised gold nanoparticles (AuNPs). Antibody functionalised nanotags (ERα-AuNPs) showed excellent biocompatibility and enabled the spatial and temporal understanding of ERα location in breast cancer cell lines with different ERα expression status. Additionally, we developed an approach based on the percentage area of SERS response to qualitatively measure expression level in ERα positive (ERα+) breast cancer cells. Specifically, the calculation of relative SERS response demonstrated that MCF-7 cells (ERα+) exhibited higher nanotag accumulation resulting in a 4.2-times increase in SERS signal area in comparison to SKBR-3 cells (ERα-). These results confirmed the strong targeting effect of ERα-AuNPs towards the ERα receptor. The functionalised ERα-AuNP nanotags were also used to investigate the activity of fulvestrant, the first-in-class approved selective estrogen receptor degrader (SERD). SERS mapping confirmed that ERα degradation occurred after fulvestrant treatment since a weaker SERS signal, and hence accumulation of nanotags, was observed in MCF-7 cells treated with fulvestrant. Most importantly, a correlation coefficient of 0.9 between the SERS response and the ERα expression level, obtained by western blot, was calculated. These results confirmed the strong relationship between the two approaches and open up the possibilities of using SERS as a tool for the estimation of ERα expression levels, without the requirement of destructive and time-consuming techniques. Therefore, the potential of using SERS as a rapid and sensitive method to understand the activity of SERDs in breast cancer is demonstrated

South Virgin-White Hills detachment fault system of SE Nevada and NW Arizona: Applying apatite fission track thermochronology to constrain the tectonic evolution of a major continental detachment fault

The South Virgin-White Hills detachment (SVWHD) in the central Basin and Range province with an along-strike extent of similar to 60 km is a major continental detachment fault system. Displacement on the SVWHD decreases north to south from similar to 17 to <6 km. This is accompanied by a change in fault and footwall rock type from mylonite overprinted by cataclasite to chlorite cataclasite and then fault breccia reflecting decreasing fault displacement and footwall exhumation. Apatite fission track (AFT) thermochronology was applied both along-strike and across-strike to assess this displacement gradient. The overall thermal history reflects Laramide cooling (similar to 75 Ma) and then rapid cooling beginning in the late early Miocene. Age patterns reflect some complexity but extension along the SVWHD appears synchronous with rapid cooling initiated at similar to 17 Ma due to tectonic exhumation. Slip rate is more rapid (similar to 8.6 km/Ma) in the north compared to similar to 1 km/Ma in the south. The displacement gradient results from penecontemporaneous along-strike motion and formation of the SVWHD by linkage of originally separate fault segments that have differential displacements and hence differential slip rates. East west transverse structures likely play a role in linkage of different fault segments. The preextension paleogeothermal gradient is well constrained in the Gold Butte block as 18-20 degrees C/km. We present a new thermochronologic approach to constrain fault dip during slip, treating the vertical exhumation rate and the slip as vectors, with the angle between them used to constrain fault dip during slip through the closure temperature of a particular thermochronometer. AFT data from the western rim of the Colorado Plateau. Citation: Fitzgerald, P. G., E. M. Duebendorfer, J. E. Faulds, and P. O'Sullivan (2009), South Virgin-White Hills detachment fault system of SE Nevada and NW Arizona: Applying apatite fission track thermochronology to constrain the tectonic evolution of a major continental detachment fault, Tectonics, 28, TC2001, doi:10.1029/2007TC002194

Detection of Estrogen Receptor Alpha and Assessment of Fulvestrant Activity in MCF-7 Tumor Spheroids Using Microfluidics and SERS

Breast cancer is one of the leading causes of cancer death in women. Novel in vitro tools that integrate three-dimensional (3D) tumor models with highly sensitive chemical reporters can provide useful information to aid biological characterization of cancer phenotype and understanding of drug activity. The combination of surface-enhanced Raman scattering (SERS) techniques with microfluidic technologies offers new opportunities for highly selective, specific, and multiplexed nanoparticle-based assays. Here, we explored the use of functionalized nanoparticles for the detection of estrogen receptor alpha (ERα) expression in a 3D tumor model, using the ERα-positive human breast cancer cell line MCF-7. This approach was used to compare targeted versus nontargeted nanoparticle interactions with the tumor model to better understand whether targeted nanotags are required to efficiently target ERα. Mixtures of targeted anti-ERα antibody-functionalized nanotags (ERα-AuNPs) and nontargeted (against ERα) anti-human epidermal growth factor receptor 2 (HER2) antibody-functionalized nanotags (HER2-AuNPs), with different Raman reporters with a similar SERS signal intensity, were incubated with MCF-7 spheroids in microfluidic devices and spectroscopically analyzed using SERS. MCF-7 cells express high levels of ERα and no detectable levels of HER2. 2D and 3D SERS measurements confirmed the strong targeting effect of ERα-AuNP nanotags to the MCF-7 spheroids in contrast to HER2-AuNPs (63% signal reduction). Moreover, 3D SERS measurements confirmed the differentiation between the targeted and the nontargeted nanotags. Finally, we demonstrated how nanotag uptake by MCF-7 spheroids was affected by the drug fulvestrant, the first-in-class approved selective estrogen receptor degrader (SERD). These results illustrate the potential of using SERS and microfluidics as a powerful in vitro platform for the characterization of 3D tumor models and the investigation of SERD activity

Surface science of soft scorpionates

The chemisorption of the soft scorpionate Li[PhTmMe] onto silver and gold surfaces is reported. Surface enhanced Raman spectroscopy in combination with the Raman analysis of suitable structural models, namely, [Cu(κ3-S,S,S-PhTmMe)(PCy3)], [Ag(κ3-S,S,S-PhTmMe)(PCy3)], [Ag(κ2-S,S-PhTmMe)(PEt3)], and [Au(κ1-S-PhTmMe)(PCy3)], are employed to identify the manner in which this potentially tridentate ligand binds to these surfaces. On colloidal silver surface-enhanced Raman spectroscopy (SERS) spectra are consistent with PhTmMe binding in a didentate fashion to the surface, holding the aryl group in close proximity to the surface. In contrast, on gold colloid, we observe that the species prefers a monodentate coordination in which the aryl group is not in close proximity to the surface

Estrogen inhibits GH signaling by suppressing GH-induced JAK2 phosphorylation, an effect mediated by SOCS-2

Oral estrogen administration attenuates the metabolic action of growth hormone (GH) in humans. To investigate the mechanism involved, we studied the effects of estrogen on GH signaling through Janus kinase (JAK)2 and the signal transducers and activators of transcription (STATs) in HEK293 cells stably expressing the GH receptor (293GHR), HuH7 (hepatoma) and T-47D (breast cancer) cells. 293GHR cells were transiently transfected with an estrogen receptor-α expression plasmid and luciferase reporters with binding elements for STAT3 and STAT5 or the β-casein promoter. GH stimulated the reporter activities by four- to sixfold. Cotreatment with 17β-estradiol (E2) resulted in a dose-dependent reduction in the response of all three reporters to GH to a maximum of 49-66% of control at 100 nM (P < 0.05). No reduction was seen when E2 was added 1-2 h after GH treatment. Similar inhibitory effects were observed in HuH7 and T-47D cells. E2 suppressed GH-induced JAK2 phosphorylation, an effect attenuated by actinomycin D, suggesting a requirement for gene expression. Next, we investigated the role of the suppressors of cytokine signaling (SOCS) in E2 inhibition. E2 increased the mRNA abundance of SOCS-2 but not SOCS-1 and SOCS-3 in HEK293 cells. The inhibitory effect of E2 was absent in cells lacking SOCS-2 but not in those lacking SOCS-1 and SOCS-3. In conclusion, estrogen inhibits GH signaling, an action mediated by SOCS-2. This paper provides evidence for regulatory interaction between a sex steroid and the GH/JAK/STAT pathway, in which SOCS-2 plays a central mechanistic role

Protein Kinase A Regulatory Subunits in Human Adipose Tissue: Decreased R2B Expression and Activity in Adipocytes From Obese Subjects

OBJECTIVE—In human adipocytes, the cAMP-dependent pathway mediates signals originating from β-adrenergic activation, thus playing a key role in the regulation of important metabolic processes, i.e., lipolysis and thermogenesis. Cyclic AMP effects are mainly mediated by protein kinase A (PKA), whose R2B regulatory isoform is the most expressed in mouse adipose tissue, where it protects against diet-induced obesity and fatty liver development. The aim of the study was to investigate possible differences in R2B expression, PKA activity, and lipolysis in adipose tissues from obese and nonobese subjects