Detection of the dipicolinic acid biomarker in Bacillus spores using Curie-point pyrolysis mass spectrometry and Fourier transform infrared spectroscopy

Thirty-six strains of aerobic endospore-forming bacteria confirmed by polyphasic taxonomic methods to belong to Bacillus amyloliquefaciens, Bacillus cereus, Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis (including Bacillus niger and Bacillus globigii), Bacillus sphaericus, and Brevi laterosporus were grown axenically on nutrient agar, and vegetative and sporulated biomasses were analyzed by Curie-point pyrolysis mass spectrometry (PyMS) and diffuse reflectance-absorbance Fourier-transform infrared spectroscopy (FT-IR). Chemometric methods based on rule induction and genetic programming were used to determine the physiological state (vegetative cells or spores) correctly, and these methods produced mathematical rules which could be simply interpreted in biochemical terms. For PyMS it was found that m/z 105 was characteristic and is a pyridine ketonium ion (C6H3ON+) obtained from the pyrolysis of dipicolinic acid (pyridine-2,6-dicarboxylic acid; DPA), a substance found in spores but not in vegetative cells; this was confirmed using pyrolysis-gas chromatography/mass spectrometry. In addition, a pyridine ring vibration at 1447-1439 cm-1 from DPA was found to be highly characteristic of spores in FT-IR analysis. Thus, although the original data sets recorded hundreds of spectral variables from whole cells simultaneously, a simple biomarker can be used for the rapid and unequivocal detection of spores of these organisms.     

Validation of a rotational coherent anti-Stokes Raman spectroscopy model for carbon dioxide using high-resolution detection in the temperature range 294-1143 K

Experiments were performed in the temperature range of 294-1143 K in pure CO(2) using high-resolution rotational coherent anti-Stokes Raman spectroscopy (CARS), in the dual-broadband approach. Experimental single-shot spectra were recorded with high spectral resolution using a single-mode Nd:YAG laser and a relay imaging lens system on the exit of a 1 m spectrometer. A theoretical rotational CARS model for CO(2) was developed for evaluation of the experimental spectra. The evaluated mean temperatures of the recorded single-shot dual-broadband rotational coherent anti-Stokes Raman spectroscopy (DB-RCARS) spectra using this model showed good agreement with thermocouple temperatures, and the relative standard deviation of evaluated single-shot temperatures was generally 2-3%. Simultaneous thermometry and relative CO(2)/N(2)-concentration measurements were demonstrated in the product gas of premixed laminar CO/air flames at atmospheric pressure. Although the model proved to be accurate for thermometry up to 1143 K, limitations were observed at flame temperatures where temperatures were overestimated and relative CO(2)/N(2) concentrations were underestimated. Potential sources for these discrepancies are discussed.     

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.     

An algorithm for real-time dosimetry in intensity-modulated radiation therapy using the radioluminescence signal from Al2O3:C

Although the radioluminescence (RL) signal from optical fibre Al(2)O(3):C dosemeters used in medical applications is essentially proportional to dose rate, the crystals used so far are imperfect in the sense that their RL sensitivity changes with accumulated dose. A computational algorithm has been developed that corrects for these sensitivity changes. We further report on a new system that effectively separates the RL signal generated in the crystal from fluorescence and Cerenkov emission generated in the optical fibre cable using a gating technique in connection with pulsed linear accelerator radiation beams. The dosimetry system has been used for dose measurements in a phantom during an intensity-modulated radiation therapy (IMRT) treatment with 6 MV photons. The RL measurement results are in excellent agreement (i.e. within 1%) with both the OSL results and the dose delivered according to the treatment planning system. RL signals from Al(2)O(3):C can be used for real-time dose rate measurements with a time resolution of approximately 0.1 s and a spatial resolution only limited by the size of the detector (<0.5 mm).     

Challenges in using probabilistic climate change information for impact assessments: an example from the water sector

Climate change impacts and adaptation assessments have traditionally adopted a scenario-based approach, which precludes an assessment of the relative risks of particular adaptation options. Probabilistic impact assessments, especially if based on a thorough analysis of the uncertainty in an impact forecast system, enable adoption of a risk-based assessment framework. However, probabilistic impacts information is conditional and will change over time. We explore the implications of a probabilistic end-to-end risk-based framework for climate impacts assessment, using the example of water resources in the Thames River, UK. We show that a probabilistic approach provides more informative results that enable the potential risk of impacts to be quantified, but that details of the risks are dependent on the approach used in the analysis.