Gating NO Release from Nitric Oxide Synthase

We have investigated the kinetics of NO escape from Geobacillus stearothermophilus nitric oxide synthase (gsNOS). Previous work indicated that NO release was gated at position 223 in mammalian enzymes; our kinetics experiments include mutants at that position along with measurements on the wild type enzyme. Employing stopped-flow UV–vis methods, reactions were triggered by mixing a reduced enzyme/N-hydroxy-l-arginine complex with an aerated buffer solution. NO release kinetics were obtained for wt NOS and three mutants (H134S, I223V, H134S/I223V). We have confirmed that wt gsNOS has the lowest NO release rate of known NOS enzymes, whether bacterial or mammalian. We also have found that steric clashes at positions 223 and 134 hinder NO escape, as judged by enhanced rates in the single mutants. The empirical rate of NO release from the gsNOS double mutant (H134/I223V) is nearly as rapid as that of the fastest mammalian enzymes, demonstrating that both positions 223 and 134 function as gates for escape of the product diatomic molecule

Electron hopping through proteins

Biological redox machines require efficient transfer of electrons and holes for function. Reactions involving multiple tunneling steps, termed “hopping,” often promote charge separation within and between proteins that is essential for energy storage and conversion. Here we show how semiclassical electron transfer theory can be extended to include hopping reactions: graphical representations (called hopping maps) of the dependence of calculated two-step reaction rate constants on driving force are employed to account for flow in a rhenium-labeled azurin mutant as well as in two structurally characterized redox enzymes, DNA photolyase and MauG. Analysis of the 35 Å radical propagation in ribonucleotide reductases using hopping maps shows that all tyrosines and tryptophans on the radical pathway likely are involved in function. We suggest that hopping maps can facilitate the design and construction of artificial photosynthetic systems for the production of fuels and other chemicals

Stem-Boring Caterpillars of Switchgrass in the Midwestern United States

Lepidopteran stem borers were collected from switchgrass, Panicum virgatum L., tillers showing symptoms of infestation at seven locations in Illinois and Iowa, with additional observations made on larval and adult activity. Blastobasis repartella (Dietz) (Coleophoridae), whose only known host is switchgrass, was common in plots grown for \u3e5 yr, whereas the polyphagous stalk borer, Papaipema nebris (Guenée) (Noctuidae), was abundant in newly established (i.e., first- and second year) switchgrass. Haimbachia albescens Capps (Crambidae) was collected from two locations in Illinois, making switchgrass the first known host for this species. Entry holes made by B. repartella and H. albescens were usually 1-2 cm above the soil surface, precluding discrimination between these species based on external appearance of damage. Although P. nebris often entered stems within 5 cm of the soil surface, they also seemed to move between stems and were the only species entering stems at heights \u3e15 cm. Adults of B. repartella were active on and above the switchgrass canopy by 2130 hours, with peak activity at ≈0230 hours. Activity of B. repartella adults seemed greatly reduced on one night with relatively cool temperatures and low wind speeds. Data from switchgrass and giant ragweed, Ambrosia trifida L., suggest P. nebris larvae move out of switchgrass during July in search of hosts with larger diameter stems, although by then hosts such as corn, Zea mays L., or Miscanthus spp. may have outgrown the potential for serious damage. However, switchgrass could contribute to greater adult populations of P. nebris if thick-stemmed hosts such as giant ragweed are not managed

Stem-Boring Caterpillars of Switchgrass in the Midwestern United States

Lepidopteran stem borers were collected from switchgrass, Panicum virgatum L., tillers showing symptoms of infestation at seven locations in Illinois and Iowa, with additional observations made on larval and adult activity. Blastobasis repartella (Dietz) (Coleophoridae), whose only known host is switchgrass, was common in plots grown for \u3e5 yr, whereas the polyphagous stalk borer, Papaipema nebris (Guenée) (Noctuidae), was abundant in newly established (i.e., first- and second year) switchgrass. Haimbachia albescens Capps (Crambidae) was collected from two locations in Illinois, making switchgrass the first known host for this species. Entry holes made by B. repartella and H. albescens were usually 1-2 cm above the soil surface, precluding discrimination between these species based on external appearance of damage. Although P. nebris often entered stems within 5 cm of the soil surface, they also seemed to move between stems and were the only species entering stems at heights \u3e15 cm. Adults of B. repartella were active on and above the switchgrass canopy by 2130 hours, with peak activity at ≈0230 hours. Activity of B. repartella adults seemed greatly reduced on one night with relatively cool temperatures and low wind speeds. Data from switchgrass and giant ragweed, Ambrosia trifida L., suggest P. nebris larvae move out of switchgrass during July in search of hosts with larger diameter stems, although by then hosts such as corn, Zea mays L., or Miscanthus spp. may have outgrown the potential for serious damage. However, switchgrass could contribute to greater adult populations of P. nebris if thick-stemmed hosts such as giant ragweed are not managed

Structural diversity in the type IV pili of multidrug-resistant Acinetobacter

Acinetobacter baumannii is a Gram-negative coccobacillus found primarily in hospital settings that has recently emerged as a source of hospital-acquired infections. A. baumannii expresses a variety of virulence factors, including type IV pili, bacterial extracellular appendages often essential for attachment to host cells. Here, we report the high resolution structures of the major pilin subunit, PilA, from three Acinetobacter strains, demonstrating thatA. baumannii subsets produce morphologically distinct type IV pilin glycoproteins. We examine the consequences of this heterogeneity for protein folding and assembly as well as host-cell adhesion by Acinetobacter. Comparisons of genomic and structural data with pilin proteins from other species of soil gammaproteobacteria suggest that these structural differences stem from evolutionary pressure that has resulted in three distinct classes of type IVa pilins, each found in multiple species

Novel Characteristics of Valveless Pumping

This study investigates the occurrence of valveless pumping in a fluidfilled system consisting of two open tanks connected by an elastic tube. We show that directional flow can be achieved by introducing a periodic pinching applied at an asymmetrical location along the tube, and that the flow direction depends on the pumping frequency. We propose a relation between wave propagation velocity, tube length, and resonance frequencies associated with shifts in the pumping direction using numerical simulations. The eigenfrequencies of the system are estimated from the linearized system, and we show that these eigenfrequencies constitute the resonance frequencies and the horizontal slope frequencies of the system; 'horizontal slope frequency' being a new concept. A simple model is suggested, explaining the effect of the gravity driven part of the oscillation observed in response to the tank and tube diameter changes. Results are partly compared with experimental findings.Art. no. 22450

Symptoms, Distribution and Abundance of the Stem-Boring Caterpillar, Blastobasis repartella (Dietz), in Switchgrass

A potential pest of switchgrass, Panicum virgatum L., was first detected in South Dakota in 2004, where death of partially emerged leaves was noted in a small proportion of tillers. Similar “dead heart” symptoms were observed in switchgrass in Illinois during 2008 and adults of a stem-boring caterpillar were collected and identified as Blastobasis repartella (Dietz). In 2009, a survey of the central United States was used to estimate the distribution and abundance of this insect. In eight northern states, B. repartella was consistently found in both cultivated plots and natural stands of switchgrass. In four southern states, B. repartella was not detected. However, because symptoms are conspicuous for a short period of time, failure to collect stem-borers on one survey date for each southern location does not necessarily define the limit of distribution for B. repartella. Sampling in four northern states showed the proportion of tillers damaged by B. repartella ranged from 1.0–7.2%. Unlike some caterpillars that feed on native grasses, it appears that the egg-laying behavior of adult moths may preclude the use of prescribed burns as an effective method to suppress this stem-boring caterpillar. As a potential pest of switchgrass planted for biomass production, near-term research needs include refining the geographic distribution of B. repartella, quantifying potential losses of switchgrass biomass, and determining whether switchgrass may be bred for resistance this and other stem-boring insects

Isolation of Escherichia coli O157:H7 from Intact Colon Fecal Samples of Swine1

Escherichia coli O157:H7 was recovered from colon fecal samples of pigs. Polymerase chain reaction confirmed two genotypes: isolates harboring the eaeA, stx1, and stx2 genes and isolates harboring the eaeA, stx1, and hly933 genes. We demonstrate that swine in the United States can harbor potentially pathogenic E. coli O157:H7

Efficient Photometric Selection of Quasars from the Sloan Digital Sky Survey: 100,000 z<3 Quasars from Data Release One

We present a catalog of 100,563 unresolved, UV-excess (UVX) quasar candidates to g=21 from 2099 deg^2 of the Sloan Digital Sky Survey (SDSS) Data Release One (DR1) imaging data. Existing spectra of 22,737 sources reveals that 22,191 (97.6%) are quasars; accounting for the magnitude dependence of this efficiency, we estimate that 95,502 (95.0%) of the objects in the catalog are quasars. Such a high efficiency is unprecedented in broad-band surveys of quasars. This ``proof-of-concept'' sample is designed to be maximally efficient, but still has 94.7% completeness to unresolved, g<~19.5, UVX quasars from the DR1 quasar catalog. This efficient and complete selection is the result of our application of a probability density type analysis to training sets that describe the 4-D color distribution of stars and spectroscopically confirmed quasars in the SDSS. Specifically, we use a non-parametric Bayesian classification, based on kernel density estimation, to parameterize the color distribution of astronomical sources -- allowing for fast and robust classification. We further supplement the catalog by providing photometric redshifts and matches to FIRST/VLA, ROSAT, and USNO-B sources. Future work needed to extend the this selection algorithm to larger redshifts, fainter magnitudes, and resolved sources is discussed. Finally, we examine some science applications of the catalog, particularly a tentative quasar number counts distribution covering the largest range in magnitude (14.2<g<21.0) ever made within the framework of a single quasar survey.Comment: 35 pages, 11 figures (3 color), 2 tables, accepted by ApJS; higher resolution paper and ASCII version of catalog available at http://sdss.ncsa.uiuc.edu/qso/nbckde