Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

A Constitutive Equation of Thermoelasticity for Solid Materials

A new constitutive equation of thermoelasticity for crystals is presented based on the interatomic potential and solid mechanics at finite temperature. Using the new constitutive equation, the calculations for crystal copper and graphene are carried out under different loading paths at different temperatures. The calculated results are in good agreement with those of the previous thermoelasticity constitutive equation based on quantum mechanics, which clearly indicates that our new constitutive equation of thermoelasticity is correct. A lot of comparisons also show that the present theory is more concise and efficient than the previous thermal stress theory in the practical application.     

Nutrient digestibility and endogenous protein losses in the foregut and small intestine of weaned dairy calves fed calf starters with conventional or enzyme-treated soybean meal

The aims of this experiment were (1) to compare the effects of a soybean meal with an enzymatic treatment (ESBM) to reduce the concentration of antinutritional factors versus a standard soybean meal (SBM) on foregut and small intestine digestion in weaned dairy calves and (2) to estimate the endogenous losses of crude protein (CP) in the small intestine. Our hypothesis was that a diet containing ESBM instead of SBM would improve ruminal and small intestine digestion and absorption of nutrients. A T-cannula was placed in the duodenum, and a second T-cannula was installed in the distal ileum of 12 Holstein calves at approximately 3 wk of age. Calves were weaned on d 42, and on d 50 they were assigned randomly to a quadruplicated 3 × 3 Latin square with 10-d periods. Digesta samples were collected on d 7 and 8 from the ileum and d 9 and 10 from the duodenum. The diets were fed for ad libitum intake and consisted of a calf starter (CS) of 20% CP with SBM as the main source of protein (CTRL), and an isonitrogenous CS with an ESBM instead of SBM (ENZT). A third diet with a low content of CP (10%) and no soy protein was fed to estimate endogenous N losses and digestibilities of test ingredients. Flows and digestibilities of nutrients were compared between CTRL and ENZT and their test ingredients (SBM vs. ESBM, respectively). Duodenal net flows of CP and total AA as well as ruminal microbial protein synthesis per kilogram of digested CP were greater, and flow of nonprotein N and CP true (corrected by endogenous and microbial flows) foregut digestibility were lower with ENZT than CTRL. The apparent small intestine digestibilities of CP and total AA were greater for ESBM than SBM, but there were no differences between the CTRL and ENZT diets. We observed no differences in digestibilities at the duodenum or ileum of starch or NDF, but true small intestine digestibilities of CP and all AA were greater with ENZT than CTRL. Total endogenous protein losses in the small intestine estimated from calves fed the low-CP with no soy protein diet were 37 ± 1.5 g of CP and 29 ± 1.4 g of AA/kg of DMI. These values may be considered the basal endogenous losses as they are similar to values obtained with the regression method, which estimates N losses when dietary N is null. Our results indicated that the inclusion of an ESBM improved the efficiency of ruminal microbial protein synthesis per digested kilogram of organic matter and CP, and increased CP and AA absorption in the small intestine despite a greater proportion of undigested dietary protein entering the duodenum.     

TRISO fuel volume fraction and homogeneity:a nondestructive characterization

A new nondestructive method to estimate the volume fraction and homogeneity of tristructural isotropic(TRISO)-coated fuel particles in fuel compacts designed for high-temperature reactors has been developed using image analysis of conventional X-radiographs. The method is demonstrated on surrogate fuel compacts containing TRISO-coated particles with kernels made of zirconium dioxide. The methodology incorporates a correction for superimposed images of TRISO particles such that a single X-ray image obtained in any one random orientation is sufficient to characterize the fuel compact in terms of volume fraction and homogeneity. The method is based on the virtual segregation of images of each particle inside the compact with the aid of a calibration standard.     

Fatal Attraction: How Bacterial Adhesins Affect Host Signaling and What We Can Learn from Them

The ability of bacterial species to colonize and infect host organisms is critically dependent upon their capacity to adhere to cellular surfaces of the host. Adherence to cell surfaces is known to be essential for the activation and delivery of certain virulence factors, but can also directly affect host cell signaling to aid bacterial spread and survival. In this review we will discuss the recent advances in the field of bacterial adhesion, how we are beginning to unravel the effects adhesins have on host cell signaling, and how these changes aid the bacteria in terms of their survival and evasion of immune responses. Finally, we will highlight how the exploitation of bacterial adhesins may provide new therapeutic avenues for the treatment of a wide range of bacterial infections.     

Real-time organic aerosol chemical speciation in the indoor environment using extractive electrospray ionization mass spectrometry

Understanding the sources and composition of organic aerosol (OA) in indoor environments requires rapid measurements, since many emissions and processes have short timescales. However, real-time molecular-level OA measurements have not been reported indoors. Here, we present quantitative measurements, at a time resolution of five seconds, of molecular ions corresponding to diverse aerosol-phase species, by applying extractive electrospray ionization mass spectrometry (EESI-MS) to indoor air analysis for the first time, as part of the highly instrumented HOMEChem field study. We demonstrate how the complex spectra of EESI-MS are screened in order to extract chemical information and investigate the possibility of interference from gas-phase semivolatile species. During experiments that simulated the Thanksgiving US holiday meal preparation, EESI-MS quantified multiple species, including fatty acids, carbohydrates, siloxanes, and phthalates. Intercomparisons with Aerosol Mass Spectrometer (AMS) and Scanning Mobility Particle Sizer suggest that EESI-MS quantified a large fraction of OA. Comparisons with FIGAERO-CIMS shows similar signal levels and good correlation, with a range of 100 for the relative sensitivities. Comparisons with SV-TAG for phthalates and with SV-TAG and AMS for total siloxanes also show strong correlation. EESI-MS observations can be used with gas-phase measurements to identify co-emitted gas- and aerosol-phase species, and this is demonstrated using complementary gas-phase PTR-MS observations.     

Ecophysiology and niche differentiation of Nitrospira-like bacteria, the key nitrite oxidizers in wastewater treatment plants.

Nitrite-oxidizing bacteria of the genus Nitrospira are key nitrifiers in wastewater treatment plants. Pure cultures of these organisms are unavailable, but cultivation-independent molecular methods make it possible to detect Nitrospira-like bacteria in environmental samples and to investigate their ecophysiology. Comprehensive screening of natural and engineered habitats and of public databases for 16S rRNA sequences of Nitrospira-like bacteria revealed a surprisingly high biodiversity in the genus Nitrospira, which comprises at least four phylogenetic sublineages. All Nitrospira-like bacteria detected in wastewater treatment plants belonged to the sublineages I and II. Subsequently, the population dynamics of different Nitrospira-like bacteria were monitored, by quantitative fluorescence in situ hybridization with rRNA-targeted probes, confocal laser scanning microscopy and digital image analysis, during incubation of nitrifying activated sludge in media containing different nitrite concentrations. These experiments showed that Nitrospira-like bacteria, which were affiliated with the phylogenetic sublineages I or II of the genus Nitrospira, responded differently to nitrite concentration shifts. Previously unknown properties of Nitrospira-like bacteria were discovered in the course of an environmental genomics project. Implications of the obtained results for fundamental understanding of the microbial ecology of nitrite oxidizers as well as for future improvement of nutrient removal in wastewater treatment plants are discussed.     

Bench- and pilot-scale sludge electrodewatering in a diaphragm filter press.

Electrodewatering is a technique in which pressure dewatering is combined with electrokinetic effects to realize an improved solid/liquid separation and hence increased filter cake dry matter contents. In order to be energy efficient, it is shown that sludge should be dewatered by pressure dewatering to a high extent prior to electric field application, and a sufficient contact time for the electric field must be guaranteed. In order to realize these goals, a bench- and pilot-scale diaphragm filter press suited for electrodewatering were constructed for treatment of sewage and other types of sludges. It was shown that electrodewatering of sludge is a feasible technique, especially for biological sludge types. Other types of sludge are less suited for electrodewatering because of the restricted improvements that can be realized in cake dry matter content and the high electric energy consumption. Furthermore, it was shown in pilot-scale tests that the use of a diaphragm filter press with electrodewatering facilities was very well suited to deliver dry filter cakes of sewage sludge at a moderate energy consumption. Depending on local market prices for investment, operating and sludge disposal costs, this technology may therefore lead to important savings in the sludge management process.     

HIGHER PLANT BIOMARKERS IN PALEOGENE CRUDE OILS FROM THE YUFUTSU OIL-AND GASFIELD AND OFFSHORE WILDCATS, JAPAN

Geochemical investigation of Paleogene oils from the onshore Yufutsu oil- and gasfield, southern Hokkaido, and from two nearby offshore wells, revealed the presence of numerous biomarkers of higher plant origin. Biomarkers in the oils belong to different groups of both angiosperm and gymnosperm origin; they include bicyclic sesquiterpanes, diterpanes, and triterpanes and their aromatized counterparts, which suggests a terrestrial origin for the oils. The oils were characterized as having a high wax content, a low content of organosulphur compounds, a high pristane/phytane ratio, and a low C₂₇/(C₂₇+C₂₉) sterane ratio.
Although the oils from on- and offshore Southern Hokkaido are similar in their geochemical composition, notable differences were observed in the biomarker signature of both saturate and aromatic fractions. The oils from the offshore wells appeared to have a greater abundance of higher plant biomarkers compared to those from the Yufutsu field, suggesting an enrichment in higher plant components. Differences in biomarker fingerprint could not be linked to the maturity effect, since the oils appeared to be of similar maturity levels, corresponding to the late stage of the oil window (0.9–1.2%, Rc). The differences in the biomarker signatures between the oils from the Yufutsu field and the offshore wells are likely to be due to facies variations in source organic matter, resulting from differences in the quantity and quality of land plant input.