High-resolution VUV spectroscopy: New results from the Advanced Light Source

Third-generation synchrotron light sources are providing photon beams of unprecedented brightness for researchers in atomic and molecular physics. Beamline 9.0.1, an undulator beamline at the Advanced Light Source (ALS), produces a beam in the vacuum-ultraviolet (VUV) region of the spectrum with exceptional flux and spectral resolution. Exciting new results from experiments in atomic and molecular VUV spectroscopy of doubly excited autoionizing states of helium, hollow lithium, and photoelectron spectroscopy of small molecules using Beamline 9.0.1 at the ALS are reported

Synthesis and biodistribution of immunoconjugates of a human IgM and polymeric drug carriers

The synthesis and purification of radiolabelled immunoconjugates, composed of a human IgM monoclonal antibody directed against an intracellular tumour-associated antigen and either poly (alpha-L-glutamic acid) (PGA) or poly[N5-(2-hydroxyethyl)-L-glutamine] (PHEG) is described. Coupling of polymers to the antibody was performed through disulfide bond formation involving a single thiol group at the C-terminus of the polymer chain and 2-pyridyldisulfide groups introduced onto the antibody. The antibody was iodinated with 131I before conjugation. The polymers contained tyrosinamide in a low degree of substitution and were radiolabelled with 125I. 125I-labelled PGA and PHEG were found to be stable for at least 3 days in murine and human plasma. The biodistribution in mice of the doubly labelled immunoconjugates was studied and was compared with the pharmacokinetics of the individual components.\ud \ud PHEG showed a relatively slow blood clearance, the half-life being approximately 10 h with low uptake in liver, kidneys and spleen. PGA was rapidly cleared from the circulation and was significantly taken up in liver, kidneys and spleen. The biodistribution of both immunoconjugates was indistinguishable from that of the IgM proper, with plasma half-lives of approximately 6 h, indicating that the pharmacokinetic properties of the immunoconjugates are largely determined by the antibody part

Synthesis and biological evaluation of immunoconjugates of adriamycin and a human IgM linked by poly[N5-(2-hydroxyethyl)-l-glutamine

The synthesis and purification of radiolabelled immunoconjugates, composed of a human IgM monoclonal antibody (IgM 16.88) directed against an intracellular tumour-associated antigen, the drug carrier poly[N5-(2-hydroxyethyl)--glutamine] (PHEG) and the cytostatic drug adriamycin (ADR) are described. The immunoconjugates were constructed to allow selective release of ADR in the putatively acidic environment of the tumour through a novel acid-labile maleamic acid linker. The conjugate of PHEG and the acid-labile ADR derivative effectively released ADR in cytotoxic amounts at a pH of 6.0 as judged from incubation in buffer and from inhibition of the growth of HT-29 colon tumour cells in vitro. Immunoconjugates were prepared by coupling of PHEG-ADR having a hydrolytically stable amide bond with 131I-labelled antibody through thioether bond formation involving a single thiol group at the C-terminus of the polymer chain and maleimido groups introduced onto th

A method to quantify FRET stoichiometry with phasor plot analysis and acceptor lifetime ingrowth.

FRET is widely used for the study of protein-protein interactions in biological samples. However, it is difficult to quantify both the FRET efficiency (E) and the affinity (Kd) of the molecular interaction from intermolecular FRET signals in samples of unknown stoichiometry. Here, we present a method for the simultaneous quantification of the complete set of interaction parameters, including fractions of bound donors and acceptors, local protein concentrations, and dissociation constants, in each image pixel. The method makes use of fluorescence lifetime information from both donor and acceptor molecules and takes advantage of the linear properties of the phasor plot approach. We demonstrate the capability of our method in vitro in a microfluidic device and also in cells, via the determination of the binding affinity between tagged versions of glutathione and glutathione S-transferase, and via the determination of competitor concentration. The potential of the method is explored with simulations.This work was funded by grants from the Medical Research Council, the Wellcome Trust, the Alzheimer Research UK Trust, and the Engineering and Physical Sciences Research Council. W.Y.C. is funded by a China Scholarship Council-Cambridge Scholarship. D.R. is a Principal Research Fellow of the Wellcome Trust.This is the final published version. It first appeared at http://www.sciencedirect.com/science/article/pii/S0006349515000752#

FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells.

BACKGROUND: During its intraerythrocytic asexual reproduction cycle Plasmodium falciparum consumes up to 80% of the host cell hemoglobin, in large excess over its metabolic needs. A model of the homeostasis of falciparum-infected red blood cells suggested an explanation based on the need to reduce the colloid-osmotic pressure within the host cell to prevent its premature lysis. Critical for this hypothesis was that the hemoglobin concentration within the host cell be progressively reduced from the trophozoite stage onwards. METHODOLOGY/PRINCIPAL FINDINGS: The experiments reported here were designed to test this hypothesis by direct measurements of the hemoglobin concentration in live, infected red cells. We developed a novel, non-invasive method to quantify the hemoglobin concentration in single cells, based on Förster resonance energy transfer between hemoglobin molecules and the fluorophore calcein. Fluorescence lifetime imaging allowed the quantitative mapping of the hemoglobin concentration within the cells. The average fluorescence lifetimes of uninfected cohorts was 270+/-30 ps (mean+/-SD; N = 45). In the cytoplasm of infected cells the fluorescence lifetime of calcein ranged from 290+/-20 ps for cells with ring stage parasites to 590+/-13 ps and 1050+/-60 ps for cells with young trophozoites and late stage trophozoite/early schizonts, respectively. This was equivalent to reductions in hemoglobin concentration spanning the range from 7.3 to 2.3 mM, in line with the model predictions. An unexpected ancillary finding was the existence of a microdomain under the host cell membrane with reduced calcein quenching by hemoglobin in cells with mature trophozoite stage parasites. CONCLUSIONS/SIGNIFICANCE: The results support the predictions of the colloid-osmotic hypothesis and provide a better understanding of the homeostasis of malaria-infected red cells. In addition, they revealed the existence of a distinct peripheral microdomain in the host cell with limited access to hemoglobin molecules indicating the concentration of substantial amounts of parasite-exported material

Deep-PowerX: A Deep Learning-Based Framework for Low-Power Approximate Logic Synthesis

This paper aims at integrating three powerful techniques namely Deep Learning, Approximate Computing, and Low Power Design into a strategy to optimize logic at the synthesis level. We utilize advances in deep learning to guide an approximate logic synthesis engine to minimize the dynamic power consumption of a given digital CMOS circuit, subject to a predetermined error rate at the primary outputs. Our framework, Deep-PowerX, focuses on replacing or removing gates on a technology-mapped network and uses a Deep Neural Network (DNN) to predict error rates at primary outputs of the circuit when a specific part of the netlist is approximated. The primary goal of Deep-PowerX is to reduce the dynamic power whereas area reduction serves as a secondary objective. Using the said DNN, Deep-PowerX is able to reduce the exponential time complexity of standard approximate logic synthesis to linear time. Experiments are done on numerous open source benchmark circuits. Results show significant reduction in power and area by up to 1.47 times and 1.43 times compared to exact solutions and by up to 22% and 27% compared to state-of-the-art approximate logic synthesis tools while having orders of magnitudes lower run-time

K-Shell Photoabsorption Studies of the Carbon Isonuclear Sequence

K-shell photoabsorption cross sections for the isonuclear C I - C IV ions have been computed using the R-matrix method. Above the K-shell threshold, the present results are in good agreement with the independent-particle results of Reilman & Manson (1979). Below threshold, we also compute the strong 1s -> np absorption resonances with the inclusion of important spectator Auger broadening effects. For the lowest 1s -> 2p, 3p resonances, comparisons to available C II, C III, and C IV experimental results show good agreement in general for the resonance strengths and positions, but unexplained discrepancies exist. Our results also provide detailed information on the C I K-shell photoabsorption cross section including the strong resonance features, since very limited laboratory experimental data exist. The resultant R-matrix cross sections are then used to model the Chandra X-ray absorption spectrum of the blazar Mkn 421