Development of an Intact Mass Spectrometry Method for the Detection and Differentiation of Major Bovine Milk Proteins

The authentication of products with claims regarding protein sources or compositions is a challenge for traditional analytical methods, which generally lack the required specificity whole protein analysis can provide. For example, the establishment of milk as “A2” is achieved through genetic testing of cows before milk production, with no methods to authenticate milk products themselves. Establishment of A2 milk is completed through genetic testing of the cows before milk production, but with no methods to authenticate the milk products themselves. Intact protein mass spectrometry (MS) has the potential to directly authenticate protein products, including specific proteoform claims. The development of an intact MS method to detect and differentiate major bovine milk proteins (αS1-, αS2-, β-, K-caseins, β-lactoglobulins, and α-lactalbumins) and their proteoforms is needed for protein profile claims and can be an effective tool to analyze milk products for protein authentication. This was attained through three major phases: generation of a predicted mass database, optimization of sample preparation and instrument parameters conducive with intact bovine milk proteins, and the selection of deconvolution software for protein identification with a mass error tolerance (10 ppm). Fifteen powdered and HTST liquid milk products with an equal distribution of marked A2 and normal commercial products were selected. Each sample was diluted to 1 mg protein/mL in 50 mM ammonium bicarbonate and then defatted through centrifugation of 15 minutes at 3,000 x g. The samples were then cleaned up, desalted through 3 kDa spin column filters, and then separated and analyzed by liquid chromatography mass spectrometry (LC-MS). Data was deconvoluted using BioPharma Finder sliding windows algorithm that were compared to the predicted database and mass were identified. A mean of 85.27% (± 6.68%, n = 57) of the total signal of powdered and liquid HTST milk could be assigned to the predicted database proteoforms using the finalized method. The average ratio of selected normal commercial products was 25.86% A1 and 0.74.14% A2. Advisors: Melanie Downs and Phillip Johnso

VisGenome: visualization of single and comparative genome representations

VisGenome visualizes single and comparative representations for the rat, the mouse and the human chromosomes at different levels of detail. The tool offers smooth zooming and panning which is more flexible than seen in other browsers. It presents information available in Ensembl for single chromosomes, as well as homologies (orthologue predictions including ortholog one2one, apparent ortholog one2one, ortholog many2many) for any two chromosomes from different species. The application can query supporting data from Ensembl by invoking a link in a browser

What is the Value of CCS in the Future Energy System?

Ambitions to produce electricity at low, zero, or negative carbon emissions are shifting the priorities and appreciation for new types of power generating technologies. Maintaining the balance between security of energy supply, carbon reduction, and electricity system cost during the transition of the electricity system is challenging. Few technology valuation tools consider the presence and interdependency of these three aspects, and nor do they appreciate the difference between firm and intermittent power generation. In this contribution, we present the results of a thought experiment and mathematical model wherein we conduct a systems analyses on the effects of gas-fired power plants equipped with Carbon Capture and Storage (CCS) technology in comparison with onshore wind power plants as main decarbonisation technologies. We find that while wind capacity integration is in its early stages of deployment an economic decarbonisation strategy, it ultimately results in an infrastructurally inefficient system with a required ratio of installed capacity to peak demand of nearly 2.. Due to the intermittent nature of wind power generation, its deployment requires a significant amount of reserve capacity in the form of firm capacity. While the integration of CCS-equipped capacity increases total system cost significantly, this strategy is able to achieve truly low-carbon power generation at 0.04 tCO2/MWh. Via a simple example, this work elucidates how the changing system requirements necessitate a paradigm shift in the value perception of power generation technologies

Evaluating RNAlater® as a preservative for using near-infrared spectroscopy to predict Anopheles gambiae age and species.

Mosquito age and species identification is a crucial determinant of the efficacy of vector control programmes. Near-infrared spectroscopy (NIRS) has previously been applied successfully to rapidly, non-destructively, and simultaneously determine the age and species of freshly anesthetized African malaria vectors from the Anopheles gambiae s.l. species complex: An. gambiae s. s. and Anopheles arabiensis. However, this has only been achieved on freshly-collected specimens and future applications will require samples to be preserved between field collections and scanning by NIRS. In this study, a sample preservation method (RNAlater(®)) was evaluated for mosquito age and species identification by NIRS against scans of fresh samples. Two strains of An. gambiae s.s. (CDC and G3) and two strains of An. arabiensis (Dongola, KGB) were reared in the laboratory while the third strain of An. arabiensis (Ifakara) was reared in a semi-field system. All mosquitoes were scanned when fresh and rescanned after preservation in RNAlater(®) for several weeks. Age and species identification was determined using a cross-validation. The mean accuracy obtained for predicting the age of young (<7 days) or old (≥ 7 days) of all fresh (n = 633) and all preserved (n = 691) mosquito samples using the cross-validation technique was 83% and 90%, respectively. For species identification, accuracies were 82% for fresh against 80% for RNAlater(®) preserved. For both analyses, preserving mosquitoes in RNAlater(®) was associated with a highly significant reduction in the likelihood of a misclassification of mosquitoes as young or old using NIRS. Important to note is that the costs for preserving mosquito specimens with RNAlater(®) ranges from 3-13 cents per insect depending on the size of the tube used and the number of specimens pooled in one tube. RNAlater(®) can be used to preserve mosquitoes for subsequent scanning and analysis by NIRS to determine their age and species with minimal costs and with accuracy similar to that achieved from fresh insects. Cold storage availability allows samples to be stored longer than a week after field collection. Further study to develop robust calibrations applicable to other strains from diverse ecological settings is recommended

A Multi-Wavelength Study of Sgr A*: The Role of Near-IR Flares in Production of X-ray, Soft γ\gamma-ray and Sub-millimeter Emission

(abridged) We describe highlights of the results of two observing campaigns in 2004 to investigate the correlation of flare activity in Sgr A* in different wavelength regimes, using a total of nine ground and space-based telescopes. We report the detection of several new near-IR flares during the campaign based on {\it HST} observations. The level of near-IR flare activity can be as low as $\sim0.15$ mJy at 1.6 $\mu$m and continuous up to about 40% of the total observing time. Using the NICMOS instrument on the {\it HST}, the {\it XMM-Newton} and CSO observatories, we also detect simultaneous bright X-ray and near-IR flare in which we observe for the first time correlated substructures as well as simultaneous submillimeter and near-IR flaring. X-ray emission is arising from the population of near-IR-synchrotron-emitting relativistic particles which scatter submillimeter seed photons within the inner 10 Schwarzschild radii of Sgr A* up to X-ray energies. In addition, using the inverse Compton scattering picture, we explain the high energy 20-120 keV emission from the direction toward Sgr A*, and the lack of one-to-one X-ray counterparts to near-IR flares, by the variation of the magnetic field and the spectral index distributions of this population of nonthermal particles. In this picture, the evidence for the variability of submillimeter emission during a near-IR flare is produced by the low-energy component of the population of particles emitting synchrotron near-IR emission. Based on the measurements of the duration of flares in near-IR and submillimeter wavelengths, we argue that the cooling could be due to adiabatic expansion with the implication that flare activity may drive an outflow.Comment: 48 pages, 12 figures, ApJ (in press

Far Infrared Variability of Sagittarius A*: 25.5 Hours of Monitoring with HerschelHerschel

Variable emission from Sgr~A*, the luminous counterpart to the super-massive black hole at the center of our Galaxy, arises from the innermost portions of the accretion flow. Better characterization of the variability is important for constraining models of the low-luminosity accretion mode powering Sgr~A*, and could further our ability to use variable emission as a probe of the strong gravitational potential in the vicinity of the $4\times10^{6}\mathrm{M}_{\odot}$ black hole. We use the \textit{Herschel} Spectral and Photometric Imaging Receiver (SPIRE) to monitor Sgr~A* at wavelengths that are difficult or impossible to observe from the ground. We find highly significant variations at 0.25, 0.35, and 0.5 mm, with temporal structure that is highly correlated across these wavelengths. While the variations correspond to $<$1% changes in the total intensity in the \textit{Herschel} beam containing Sgr~A*, comparison to independent, simultaneous observations at 0.85 mm strongly supports the reality of the variations. The lowest point in the light curves, $\sim$0.5 Jy below the time-averaged flux density, places a lower bound on the emission of Sgr~A* at 0.25 mm, the first such constraint on the THz portion of the SED. The variability on few hour timescales in the SPIRE light curves is similar to that seen in historical 1.3 mm data, where the longest time series is available, but the distribution of variations in the sub-mm do not show a tail of large-amplitude variations seen at 1.3 mm. Simultaneous X-ray photometry from XMM-Newton shows no significant variation within our observing period, which may explain the lack of very large variations if X-ray and submillimeter flares are correlated.Comment: Accepted for publication in Ap

Flaring Activity of Sgr A*: Expanding Hot Blobs

Sgr A* is considered to be a massive black hole at the Galactic center and is known to be variable in radio, millimeter, near-IR and X-rays. Recent multi-wavelength observing campaigns show a simultaneous X-ray and near-IR flare, as well as sub-millimeter and near-IR flares from Sgr A*. The flare activity is thought to be arising from the innermost region of Sgr A*. We have recently argued that the duration of flares in near-IR and submillimeter wavelengths implies that the burst of emission expands and cools on a dynamical time scale before the flares leave Sgr A*. The detection of radio flares with a time delay in the range of 20 and 40 minutes between 7 and 12mm peak emission implies adiabatic expansion of a uniform, spherical hot blob due to flare activity. We suspect that this simple outflow picture shows some of the characteristics that are known to take place in microquasars, thus we may learn much from comparative study of Sgr A* and its environment vs. microquasars.Comment: 10 pages, 5 figures, to be published in IV Microquasar Workshop: Microquasars and Beyond, September 18-22 2006, Como, Ital