65 research outputs found

    Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the alpha subunit of bacterial luciferase

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    The nucleotide sequence of the 1.85-kilobase EcoRI fragment from Vibrio harveyi that was cloned using a mixed-sequence synthetic oligonucleotide probe (Cohn, D. H., Ogden, R. C., Abelson, J. N., Baldwin, T. O., Nealson, K. H., Simon, M. I., and Mileham, A. J. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 120-123) has been determined. The alpha subunit-coding region (luxA) was found to begin at base number 707 and end at base number 1771. The alpha subunit has a calculated molecular weight of 40,108 and comprises a total of 355 amino acid residues. There are 34 base pairs separating the start of the alpha subunit structural gene and a 669-base open reading frame extending from the proximal EcoRI site. At the 3' end of the luxA coding region there are 26 bases between the end of the structural gene and the start of the luxB structural gene. Approximately two-thirds of the alpha subunit was sequenced by protein chemical techniques. The amino acid sequence implied by the DNA sequence, with few exceptions, confirmed the chemically determined sequence. Regions of the alpha subunit thought to comprise the active center were found to reside in two discrete and relatively basic regions, one from around residues 100-115 and the second from around residues 280-295

    Current advances on Talbot–Lau x-ray imaging diagnostics for high energy density experiments (invited)

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    Producción CientíficaTalbot–Lau x-ray interferometry is a refraction-based diagnostic that can map electron density gradients through phase-contrast methods. The Talbot–Lau x-ray deflectometry (TXD) diagnostics have been deployed in several high energy density experiments. To improve diagnostic performance, a monochromatic TXD was implemented on the Multi-Tera Watt (MTW) laser using 8 keV multilayer mirrors (Δθ/θ = 4.5%-5.6%). Copper foil and wire targets were irradiated at 1014–1015 W/cm2. Laser pulse length (∼10 to 80 ps) and backlighter target configurations were explored in the context of Moiré fringe contrast and spatial resolution. Foil and wire targets delivered increased contrast <30%. The best spatial resolution (<6 μm) was measured for foils irradiated 80° from the surface. Further TXD diagnostic capability enhancement was achieved through the development of advanced data postprocessing tools. The Talbot Interferometry Analysis (TIA) code enabled x-ray refraction measurements from the MTW monochromatic TXD. Additionally, phase, attenuation, and dark-field maps of an ablating x-pinch load were retrieved through TXD. The images show a dense wire core of ∼60 μm diameter surrounded by low-density material of ∼40 μm thickness with an outer diameter ratio of ∼2.3. Attenuation at 8 keV was measured at ∼20% for the dense core and ∼10% for the low-density material. Instrumental and experimental limitations for monochromatic TXD diagnostics are presented. Enhanced postprocessing capabilities enabled by TIA are demonstrated in the context of high-intensity laser and pulsed power experimental data analysis. Significant advances in TXD diagnostic capabilities are presented. These results inform future diagnostic technique upgrades that will improve the accuracy of plasma characterization through TXD

    Ramucirumab plus erlotinib in patients with untreated, EGFR-mutated, advanced non-small-cell lung cancer (RELAY): a randomised, double-blind, placebo-controlled, phase 3 trial

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    Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function

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    Porcine Reproductive and Respiratory Syndrome (PRRS) is a panzootic infectious disease of pigs, causing major economic losses to the world-wide pig industry. PRRS manifests differently in pigs of all ages but primarily causes late-term abortions and stillbirths in sows and respiratory disease in piglets. The causative agent of the disease is the positive-strand RNA PRRS virus (PRRSV). PRRSV has a narrow host cell tropism, limited to cells of the monocyte/macrophage lineage. CD163 has been described as a fusion receptor for PRRSV, whereby the scavenger receptor cysteine-rich domain 5 (SRCR5) region was shown to be an interaction site for the virus in vitro. CD163 is expressed at high levels on the surface of macrophages, particularly in the respiratory system. Here we describe the application of CRISPR/Cas9 to pig zygotes, resulting in the generation of pigs with a deletion of Exon 7 of the CD163 gene, encoding SRCR5. Deletion of SRCR5 showed no adverse effects in pigs maintained under standard husbandry conditions with normal growth rates and complete blood counts observed. Pulmonary alveolar macrophages (PAMs) and peripheral blood monocytes (PBMCs) were isolated from the animals and assessed in vitro. Both PAMs and macrophages obtained from PBMCs by CSF1 stimulation (PMMs) show the characteristic differentiation and cell surface marker expression of macrophages of the respective origin. Expression and correct folding of the SRCR5 deletion CD163 on the surface of macrophages and biological activity of the protein as hemoglobin-haptoglobin scavenger was confirmed. Challenge of both PAMs and PMMs with PRRSV genotype 1, subtypes 1, 2, and 3 and PMMs with PRRSV genotype 2 showed complete resistance to viral infections assessed by replication. Confocal microscopy revealed the absence of replication structures in the SRCR5 CD163 deletion macrophages, indicating an inhibition of infection prior to gene expression, i.e. at entry/fusion or unpacking stages

    Longitudinal study of serum thyroid hormone levels during normal pregnancy

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    We undertook a prospective longitudinal study of thyroid function in 60 normal pregnant women and measured serum concentrations of T 4, triiodothyronine (T 3), T-uptake, thyroxine binding globulin (TBG), free thyroxine index (FTI), free T 4, albumin and thyrotropin (TSH). From these data we established reference ranges for each of these analytes for each trimester and examined the interrelationships between laboratory measurements of thyroid function tests. We observed significant increases in serum concentrations of thyrotropin and decreases in free T 4, assays commonly used as first line investigations of thyroid activity during pregnancy. However, the 95th centile intervals for both analytes remained within the reference range for nonpregnant women

    Chromosomal mapping of the mink cell focus-inducing and xenotropic env gene family in the mouse.

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    Chromosomal locations of members of the xenotropic-related env gene family in the mouse genome have been determined. Endonuclease restriction site polymorphisms detected by molecular hybridization were used to study the inheritance of mink cell-focus inducing and xenotropic env gene-related sequences in recombinant inbred strains of mice. Some of the endogenous env sequences appear to be closely linked to genes determining leukemia virus induction and to genes involved in the immune response, such as the heavy and light chains of the immunoglobulin molecules or allotypic determinants on B and T lymphocytes. The use of probes that detect restriction fragment length polymorphisms in a small family of dispersed sequences promises to yield a large number of markers that can be used together with recombinant inbred strains for efficient mapping of the mouse genome
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