Extracellular ATP mediates proinflammatory and antiproliferative effects via activation of P2 nucleotide receptors. In contrast, its metabolite, the nucleoside adenosine, is strongly immunosuppressive and enhances tumor proliferation and metastasis. The conversion of ATP to adenosine is catalyzed by ectonucleotidases, which are expressed on immune cells and typically upregulated on tumor cells. In the present study, we identified sulfopolysaccharides from brown and red sea algae to act as potent dual inhibitors of the main ATP-hydrolyzing ectoenzymes, ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) and ecto-nucleoside triphosphate diphosphohydrolase-1 (NTPDase1, CD39), showing nano- to picomolar potency and displaying a non-competitive mechanism of inhibition. We showed that one of the sulfopolysaccharides tested as a representative example reduced adenosine formation at the surface of the human glioblastoma cell line U87 in a concentration-dependent manner. These natural products represent the most potent inhibitors of extracellular ATP hydrolysis known to date and have potential as novel therapeutics for the immunotherapy of cancer.
We present a Raman spectroscopy setup containing a conical beam shaper in combination with a freeform segmented reflector for surface enhanced Raman scattering (SERS) analysis. The freeform segmented reflector and the conical beam shaper are designed by numerical approaches and fabricated by means of ultra-precision diamond tooling. The segmented reflector has a numerical aperture of 0.984 and a working distance of 1mm for SERS measurements. We perform systematic simulations using non-sequential ray tracing to assess the detecting abilities of the designed SERS-based system. We implement a proof-of-concept setup and demonstrate the confocal behavior by measuring the SERS signal of 10µM rhodamine B solution. The experimental results agree well with the simulations concerning the misalignment tolerances of the beam shaper with respect to the segmented reflector and the misalignment tolerances of the collecting fiber. In addition, we conduct benchmark SERS measurements by using a 60× objective lens with a numerical aperture of 0.85. We find that the main Raman intensity of rhodamine B at 1502 cm-1 obtained by our segmented reflector working together with the conical beam shaper is approximately 30% higher compared to the commercial objective lens.
Four high resolution negative electron beam resists are compared: TEBN-1 from Tokuyama Corp. Japan, ma-N 2401XP and mr-L 6000.1XP from microresist technology GmbH Germany, and SU-8 2000 series from MicroChem Corp., USA. Narrow linewidth high density patterns are defined by 100kV electron beam lithography, and the pattern is transferred into silicon by a highly anisotropic SF6∕O2∕CHF3 based reactive ion etch process with a selectivity between silicon and the investigated resists of approximately 2. 20nm half-pitch lines and 10nm lines with a pitch down to 60nm are written and transferred into silicon.
The number of newly developed genetic variants of microbial cell factories for production of biochemicals has been rapidly growing in recent years, leading to an increased need for new screening techniques. We developed a method based on surface-enhanced Raman scattering (SERS) coupled with liquid-liquid extraction (LLE) for quantification of p-coumaric acid (pHCA) in the supernatant of genetically engineered Escherichia coli (E. coli) cultures. pHCA was measured in a dynamic range from 1 μM up to 50 μM on highly uniform SERS substrates based on leaning gold-capped nanopillars, which showed an in-wafer signal variation of only 11.7%. LLE using dichloromethane as organic phase was combined with the detection in order to increase selectivity and sensitivity by decreasing the effect of interfering compounds from the analytes of interest. The difference in pHCA production yield between three genetically engineered E. coli strains was successfully evaluated using SERS and confirmed with high-performance liquid chromatography. As this novel approach has potential to be automated and parallelized, it can be considered for high-throughput screening in metabolic engineering.
We present a nanostructured surface, made of dry etched black silicon, which lowers the reflectance for light incident at all angles. This surface is fabricated on infrared detectors used for tissue oximetry, where the detection of weak diffuse light signals is important. Monte Carlo simulations performed on a model of a neonatal head shows that approximately 60% of the injected light will be diffuse reflected. However, the change in diffuse reflected light due to the change in cerebral oxygenation is very low and the light will be completely isotropic scattered. The reflectance of the black silicon surface was measured for different angels of incident and was fund to be below 10% for angles of incident up to 70o. The quantum efficiency of detectors with the black silicon nanostructures was measured and compared to detectors with a simple anti-reflection coating. The result was an improvement in quantum efficiency for both normal incident light and light incident at 38o.
Multiple independent sensors are used in security and military applications in order to increase sensitivity, selectivity and data reliability. The Xsense project has been initiated at the Technical University of Denmark in collaboration with a number of partners in an effort to produce a handheld sensor for trace detection of explosives. We are using micro- and nano technological approaches for integrating four sensing principles into a single device. At the end of the project, the consortium aims at having delivered a sensor platform consisting of four independent detector principles capable of identifying concentrations of TNT, DNT, HMX and RDX at sub parts-per-billion (ppb) levels and with a false positive rate less than 1 parts-per-thousand. The specificity, sensitivity, reliability and the speed of responses are ensured by the use of advanced data processing, surface functionalization and nanostructured sensors and sensor design.
