Effects of Time of Deoxyribonucleic Acid Microinjection on Gene Detection and In Vitro Development of Bovine Embryos

In vivo fertilized embryos were surgically collected from superovulated dairy cows to evaluate microinjection on embryo development and utilized the polymerase chain reaction technique for selection of transgenic embryos. Seventy-two percent of the embryos with visible pronuclei or nuclei were microinjected with DNA, and the remaining 28% served as uninjected controls. All embryos were cocultured with bovine oviductal epithelial cells. Mean final development scores of embryos within the same initial cell stage at collection were unaffected by microinjection. After 144 h of culture, 45% of the microinjected embryos developed to the morula or blastocyst stage. The transgene was detected in 50, 10, and 9% of demimorulae from embryos microinjected at the 1-, 2-, and 4-cell stages. Frequency of transgene detection was higher in morulae from 1-cell embryos than in morulae from 2- and 4-cell embryos. Use of in vitro coculture, embryo bisection, and polymerase chain reaction technique facilitated selection of bovine embryos that carried the transgene

MetWAMer: eukaryotic translation initiation site prediction

<p>Abstract</p> <p>Background</p> <p>Translation initiation site (TIS) identification is an important aspect of the gene annotation process, requisite for the accurate delineation of protein sequences from transcript data. We have developed the MetWAMer package for TIS prediction in eukaryotic open reading frames of non-viral origin. MetWAMer can be used as a stand-alone, third-party tool for post-processing gene structure annotations generated by external computational programs and/or pipelines, or directly integrated into gene structure prediction software implementations.</p> <p>Results</p> <p>MetWAMer currently implements five distinct methods for TIS prediction, the most accurate of which is a routine that combines weighted, signal-based translation initiation site scores and the contrast in coding potential of sequences flanking TISs using a perceptron. Also, our program implements clustering capabilities through use of the <it>k</it>-medoids algorithm, thereby enabling cluster-specific TIS parameter utilization. In practice, our static weight array matrix-based indexing method for parameter set lookup can be used with good results in data sets exhibiting moderate levels of 5'-complete coverage.</p> <p>Conclusion</p> <p>We demonstrate that improvements in statistically-based models for TIS prediction can be achieved by taking the class of each potential start-methionine into account pending certain testing conditions, and that our perceptron-based model is suitable for the TIS identification task. MetWAMer represents a well-documented, extensible, and freely available software system that can be readily re-trained for differing target applications and/or extended with existing and novel TIS prediction methods, to support further research efforts in this area.</p

Autism as a disorder of neural information processing: directions for research and targets for therapy

The broad variation in phenotypes and severities within autism spectrum disorders suggests the involvement of multiple predisposing factors, interacting in complex ways with normal developmental courses and gradients. Identification of these factors, and the common developmental path into which theyfeed, is hampered bythe large degrees of convergence from causal factors to altered brain development, and divergence from abnormal brain development into altered cognition and behaviour. Genetic, neurochemical, neuroimaging and behavioural findings on autism, as well as studies of normal development and of genetic syndromes that share symptoms with autism, offer hypotheses as to the nature of causal factors and their possible effects on the structure and dynamics of neural systems. Such alterations in neural properties may in turn perturb activity-dependent development, giving rise to a complex behavioural syndrome many steps removed from the root causes. Animal models based on genetic, neurochemical, neurophysiological, and behavioural manipulations offer the possibility of exploring these developmental processes in detail, as do human studies addressing endophenotypes beyond the diagnosis itself

Post-depositional fracturing and subsidence of pumice flow deposits: Lascar Volcano, Chile

Unconsolidated pyroclastic flow deposits of the 1993 eruption of Lascar Volcano, Chile, have, with time, become increasingly dissected by a network of deeply penetrating fractures. The fracture network comprises orthogonal sets of decimeter-wide linear voids that form a pseudo-polygonal grid visible on the deposit surface. In this work, we combine shallow surface geophysical imaging tools with remote sensing observations and direct field measurements of the deposit to investigate these fractures and their underlying causal mechanisms. Based on ground penetrating radar images, the fractures are observed to have propagated to depths of up to 10 m. In addition, orbiting radar interferometry shows that deposit subsidence of up to 1 cm/year occurred between 1993 and 1996 with continued subsidence occurring at a slower rate thereafter. In situ measurements show that 1 m below the surface, the 1993 deposits remain 5°C to 15°C hotter, 18 years after emplacement, than adjacent deposits. Based on the observed subsidence as well as estimated cooling rates, the fractures are inferred to be the combined result of deaeration, thermal contraction, and sedimentary compaction in the months to years following deposition. Significant environmental factors, including regional earthquakes in 1995 and 2007, accelerated settling at punctuated moments in time. The spatially variable fracture pattern relates to surface slope and lithofacies variations as well as substrate lithology. Similar fractures have been reported in other ignimbrites but are generally exposed only in cross section and are often attributed to formation by external forces. Here we suggest that such interpretations should be invoked with caution, and deformation including post-emplacement subsidence and fracturing of loosely packed ash-rich deposits in the months to years postemplacement is a process inherent in the settling of pyroclastic material

Periodic density functional theory calculations of bulk and the (010) surface of goethite

<p>Abstract</p> <p>Background</p> <p>Goethite is a common and reactive mineral in the environment. The transport of contaminants and anaerobic respiration of microbes are significantly affected by adsorption and reduction reactions involving goethite. An understanding of the mineral-water interface of goethite is critical for determining the molecular-scale mechanisms of adsorption and reduction reactions. In this study, periodic density functional theory (DFT) calculations were performed on the mineral goethite and its (010) surface, using the Vienna <it>Ab Initio </it>Simulation Package (VASP).</p> <p>Results</p> <p>Calculations of the bulk mineral structure accurately reproduced the observed crystal structure and vibrational frequencies, suggesting that this computational methodology was suitable for modeling the goethite-water interface. Energy-minimized structures of bare, hydrated (one H₂O layer) and solvated (three H₂O layers) (010) surfaces were calculated for 1 × 1 and 3 × 3 unit cell slabs. A good correlation between the calculated and observed vibrational frequencies was found for the 1 × 1 solvated surface. However, differences between the 1 × 1 and 3 × 3 slab calculations indicated that larger models may be necessary to simulate the relaxation of water at the interface. Comparison of two hydrated surfaces with molecularly and dissociatively adsorbed H₂O showed a significantly lower potential energy for the former.</p> <p>Conclusion</p> <p>Surface Fe-O and (Fe)O-H bond lengths are reported that may be useful in surface complexation models (SCM) of the goethite (010) surface. These bond lengths were found to change significantly as a function of solvation (i.e., addition of two extra H₂O layers above the surface), indicating that this parameter should be carefully considered in future SCM studies of metal oxide-water interfaces.</p

Residual Cooling and Persistent Star Formation amid AGN Feedback in Abell 2597

New Chandra X-ray and Herschel FIR observations enable a multiwavelength study of active galactic nucleus (AGN) heating and intracluster medium (ICM) cooling in the brightest cluster galaxy of Abell 2597. The new Chandra observations reveal the central < 30 kiloparsec X-ray cavity network to be more extensive than previously thought, and associated with enough enthalpy to theoretically inhibit the inferred classical cooling flow. Nevertheless, we present new evidence, consistent with previous results, that a moderately strong residual cooling flow is persisting at 4%-8% of the classically predicted rates in a spatially structured manner amid the feedback-driven excavation of the X-ray cavity network. New Herschel observations are used to estimate warm and cold dust masses, a lower-limit gas-to-dust ratio, and a star formation rate consistent with previous measurements. The cooling time profile of the ambient X-ray atmosphere is used to map the locations of the observational star formation entropy threshold as well as the theoretical thermal instability threshold. Both lie just outside the < 30 kpc central region permeated by X-ray cavities, and star formation as well as ionized and molecular gas lie interior to both. The young stars are distributed in an elongated region that is aligned with the radio lobes, and their estimated ages are both younger and older than the X-ray cavity network, suggesting both jet-triggered as well as persistent star formation over the current AGN feedback episode. Bright X-ray knots that are coincident with extended Ly-alpha and FUV continuum filaments motivate a discussion of structured cooling from the ambient hot atmosphere along a projected axis that is perpendicular to X-ray cavity and radio axis. We conclude that the cooling ICM is the dominant contributor of the cold gas reservoir fueling star formation and AGN activity in the Abell 2597 BCG.Comment: 20 pages, 7 figures; Accepted for publication in MNRA

Juvenile Hormone (JH) Esterase of the Mosquito Culex quinquefasciatus Is Not a Target of the JH Analog Insecticide Methoprene

Juvenile hormones (JHs) are essential sesquiterpenes that control insect development and reproduction. JH analog (JHA) insecticides such as methoprene are compounds that mimic the structure and/or biological activity of JH. In this study we obtained a full-length cDNA, cqjhe, from the southern house mosquito Culex quinquefasciatus that encodes CqJHE, an esterase that selectively metabolizes JH. Unlike other recombinant esterases that have been identified from dipteran insects, CqJHE hydrolyzed JH with specificity constant (kcat/KM ratio) and Vmax values that are common among JH esterases (JHEs). CqJHE showed picomolar sensitivity to OTFP, a JHE-selective inhibitor, but more than 1000-fold lower sensitivity to DFP, a general esterase inhibitor. To our surprise, CqJHE did not metabolize the isopropyl ester of methoprene even when 25 pmol of methoprene was incubated with an amount of CqJHE that was sufficient to hydrolyze 7,200 pmol of JH to JH acid under the same assay conditions. In competition assays in which both JH and methoprene were available to CqJHE, methoprene did not show any inhibitory effects on the JH hydrolysis rate even when methoprene was present in the assay at a 10-fold higher concentration relative to JH. Our findings indicated that JHE is not a molecular target of methoprene. Our findings also do not support the hypothesis that methoprene functions in part by inhibiting the action of JHE

Customer emotions in service failure and recovery encounters

Emotions play a significant role in the workplace, and considerable attention has been given to the study of employee emotions. Customers also play a central function in organizations, but much less is known about customer emotions. This chapter reviews the growing literature on customer emotions in employee–customer interfaces with a focus on service failure and recovery encounters, where emotions are heightened. It highlights emerging themes and key findings, addresses the measurement, modeling, and management of customer emotions, and identifies future research streams. Attention is given to emotional contagion, relationships between affective and cognitive processes, customer anger, customer rage, and individual differences

Origin and ascent history of unusually crystal-rich alkaline basaltic magmas from the western Pannonian Basin

The last eruptions of the monogenetic Bakony-Balaton Highland Volcanic Field (western Pannonian Basin, Hungary) produced unusually crystal- and xenolith-rich alkaline basalts which are unique among the alkaline basalts of the Carpathian- Pannonian Region. Similar alkaline basalts are only rarely known in other volcanic fields of the world. These special basaltic magmas fed the eruptions of two closely located volcanic centres: the Bondoró-hegy and the Füzes-tó scoria cone. Their uncommon enrichment in diverse crystals produced unique rock textures and modified original magma compositions (13.1-14.2 wt.% MgO, 459-657 ppm Cr, 455-564 ppm Ni contents). Detailed mineral-scale textural and chemical analyses revealed that the Bondoró-hegy and Füzes-tó alkaline basaltic magmas have a complex ascent history, and that most of their minerals (~30 vol.% of the rocks) represent foreign crystals derived from different levels of the underlying lithosphere. The most abundant xenocrysts, olivine, orthopyroxene, clinopyroxene and spinel, were incorporated from different regions and rock types of the subcontinental lithospheric mantle. Megacrysts of clinopyroxene and spinel could have originated from pegmatitic veins / sills which probably represent magmas crystallized near the crust-mantle boundary. Green clinopyroxene xenocrysts could have been derived from lower crustal mafic granulites. Minerals that crystallized in situ from the alkaline basaltic melts (olivine with Cr-spinel inclusions, clinopyroxene, plagioclase, Fe-Ti oxides) are only represented by microphenocrysts and overgrowths on the foreign crystals. The vast amount of peridotitic (most common) and mafic granulitic materials indicates a highly effective interaction between the ascending magmas and wall rocks at lithospheric mantle and lower crustal levels. However, fragments from the middle and upper crust are absent from the studied basalts, suggesting a change in the style (and possibly rate) of magma ascent in the crust. These xenocryst- and xenolith-rich basalts yield divers tools for estimating magma ascent rate that is important for hazard forecasting in monogenetic volcanic fields. According to the estimated ascent rates, the Bondoró-hegy and Füzes-tó alkaline basaltic magmas could have reached the surface within hours to few days, similarly to the estimates for other eruptive centres in the Pannonian Basin which were fed by "normal" (crystal- and xenolith-poor) alkaline basalts