The bacterium Clostridium perfringens causes severe, sometimes lethal gastrointestinal disorders in humans, including enteritis and enterotoxemia. Type F strains produce an enterotoxin (CpE) that causes the third most common foodborne illness in the United States. CpE induces gut breakdown by disrupting barriers at cell-cell contacts called tight junctions (TJs), which are formed and maintained by claudins. Targeted binding of CpE to specific claudins, encoded by its C-terminal domain (cCpE), loosens TJ barriers to trigger molecular leaks between cells. Cytotoxicity results from claudin-bound CpE complexes forming pores in cell membranes. In mammalian tissues, ∼24 claudins govern TJ barriers-but the basis for CpE's selective targeting of claudins in the gut was undetermined. We report the structure of human claudin-4 in complex with cCpE, which reveals that enterotoxin targets a motif conserved in receptive claudins and how the motif imparts high-affinity CpE binding to these but not other subtypes. The structural basis of CpE targeting is supported by binding affinities, kinetics, and half-lives of claudin-enterotoxin complexes and by the cytotoxic effects of CpE on claudin-expressing cells. By correlating the binding residence times of claudin-CpE complexes we determined to claudin expression patterns in the gut, we uncover that the primary CpE receptors differ in mice and humans due to sequence changes in the target motif. These findings provide the molecular and structural element CpE employs for subtype-specific targeting of claudins during pathogenicity of C. perfringens in the gut and a framework for new strategies to treat CpE-based illnesses in domesticated mammals and humans.

Feline infectious peritonitis (FIP) is a highly fatal disease caused by a virulent feline coronavirus in domestic and wild cats. We have previously reported the synthesis of potent coronavirus 3C-like protease (3CLpro) inhibitors and the efficacy of a protease inhibitor, GC376, in client-owned cats with FIP. In this study, we studied the effect of the amino acid changes in 3CLpro of feline coronavirus from a feline patient who received antiviral treatment for prolonged duration. We generated recombinant 3CLpro containing the identified amino acid changes (N25S, A252S or K260 N) and determined their susceptibility to protease inhibitors in the fluorescence resonance energy transfer assay. The assay showed that N25S in 3CLpro confers a small change (up to 1.68-fold increase in the 50% inhibitory concentration) in susceptibility to GC376, but other amino acid changes do not affect susceptibility. Modelling of 3CLpro carrying the amino acid changes was conducted to probe the structural basis for these findings. The results of this study may explain the observed absence of clinical resistance to the long-term antiviral treatment in the patients.

Purpose

To compare the diagnostic performance of dual-energy (DE) computed tomography (CT) with two simultaneously administered contrast agents (hereafter, dual contrast) with that of conventional CT in the evaluation of the presence and source of extravasation in penetrating abdominopelvic trauma.

Materials and methods

Institutional animal care and use committee approval was obtained, and the study was performed in accordance with National Institutes of Health guidelines for the care and use of laboratory animals. Five rabbits with bowel trauma, vascular penetrating trauma, or both were imaged with simultaneous iodinated intravenous and bismuth subsalicylate enteric contrast material at DE CT. Four attending radiologists and six radiology residents without prior DE CT experience each evaluated 10 extraluminal collections to identify the vascular and/or enteric origin of extravasation and assess their level of diagnostic confidence, first with virtual monochromatic images simulating conventional CT and then with DE CT material decomposition attenuation maps.

Results

Overall accuracy of identification of source of extravasation increased from 78% with conventional CT to 92% with DE CT (157 of 200 diagnoses vs 184 of 200 diagnoses, respectively; P < .001). Nine radiologists were more accurate with DE CT; one had no change. Mean confidence increased from 67% to 81% with DE CT (P < .001).

Conclusion

In a rabbit abdominopelvic trauma model, dual-contrast DE CT significantly increased accuracy and confidence in the diagnosis of vascular versus enteric extravasated contrast material.

Purpose

To evaluate the feasibility of using a commercially available clinical dual-energy computed tomographic (CT) scanner to differentiate the in vivo enhancement due to two simultaneously administered contrast media with complementary x-ray attenuation ratios.

Materials and methods

Approval from the institutional animal care and use committee was obtained, and National Institutes of Health guidelines for the care and use of laboratory animals were observed. Dual-energy CT was performed in a set of iodine and tungsten solution phantoms and in a rabbit in which iodinated intravenous and bismuth subsalicylate oral contrast media were administered. In addition, a second rabbit was studied after intravenous administration of iodinated and tungsten cluster contrast media. Images were processed to produce virtual monochromatic images that simulated the appearance of conventional single-energy scans, as well as material decomposition images that separate the attenuation due to each contrast medium.

Results

Clear separation of each of the contrast media pairs was seen in the phantom and in both in vivo animal models. Separation of bowel lumen from vascular contrast medium allowed visualization of bowel wall enhancement that was obscured by intraluminal bowel contrast medium on conventional CT scans. Separation of two vascular contrast media in different vascular phases enabled acquisition of a perfectly coregistered CT angiogram and venous phase-enhanced CT scan simultaneously in a single examination.

Conclusion

Commercially available clinical dual-energy CT scanners can help differentiate the enhancement of selected pairs of complementary contrast media in vivo.

To assess the ability of dual-energy CT (DECT) to separate intravenous contrast of bowel wall from intraluminal contrast, we scanned 16 rabbits on a clinical DECT scanner: n = 3 using only iodinated intravenous contrast, and n = 13 double-contrast enhanced scans using iodinated intravenous contrast and experimental enteric non-iodinated contrast agents in the bowel lumen (five bismuth, four tungsten, and four tantalum based). Representative image pairs from conventional CT images and DECT iodine density maps of small bowel (116 pairs from 232 images) were viewed by four abdominal imaging attending radiologists to independently score each comparison pair on a visual analog scale (-100 to +100%) for (1) preference in small bowel wall visualization and (2) preference in completeness of intraluminal enteric contrast subtraction. Median small bowel wall visualization was scored 39 and 42 percentage points (95% CI 30-44% and 36-45%, both p < 0.001) higher for double-contrast DECT than for conventional CT with enteric tungsten and tantalum contrast, respectively. Median small bowel wall visualization for double-contrast DECT was scored 29 and 35 percentage points (95% CI 20-35% and 33-39%, both p < 0.001) higher with enteric tungsten and tantalum, respectively, than with bismuth contrast. Median completeness of intraluminal enteric contrast subtraction in double-contrast DECT iodine density maps was scored 28 and 29 percentage points (95% CI 15-31% and 28-33%, both p < 0.001) higher with enteric tungsten and tantalum, respectively, than with bismuth contrast. Results suggest that in vivo double-contrast DECT with iodinated intravenous and either tantalum- or tungsten-based enteric contrast provides better visualization of small bowel than conventional CT. Copyright © 2016 John Wiley & Sons, Ltd.

C-alkylpyrogallol[4]arenes (PgCs) have been studied for their ability to form metal-organic nanocapsules (MONCs) through coordination to appropriate metal ions. Here we present the synthesis and characterization of an Mn^II/Mn^III-seamed MONC in addition to its electrochemical and magnetic behavior. This MONC assembles from 24 manganese ions and 6 PgCs, while an additional metal ion is located on the capsule interior, anchored through the introduction of bridging nitrite ions. The latter originate from an in situ redox reaction that occurs during the self-assembly process, thus representing a new route to otherwise unobtainable nanocapsules.

In our quest to understand the nature of dark matter and discover its non-gravitational interactions with ordinary matter, we propose an experiment using a \pbo ~calorimeter to search for or set new limits on the production rate of i) hidden sector particles in the $3 - 60$ MeV mass range via their $e^+e^-$ decay (or $\gamma\gamma$ decay with limited tracking), and ii) the hypothetical X17 particle, claimed in multiple recent experiments. The search for these particles is motivated by new hidden sector models and dark matter candidates introduced to account for a variety of experimental and observational puzzles: the small-scale structure puzzle in cosmological simulations, anomalies such as the 4.2$\sigma$ disagreement between experiments and the standard model prediction for the muon anomalous magnetic moment, and the excess of $e^+e^-$ pairs from the $^8$Be M1 and $^4$He nuclear transitions to their ground states observed by the ATOMKI group. In these models, the $1 - 100$ MeV mass range is particularly well-motivated and the lower part of this range still remains unexplored. Our proposed direct detection experiment will use a magnetic-spectrometer-free setup (the PRad apparatus) to detect all three final state particles in the visible decay of a hidden sector particle allowing for an effective control of the background and will cover the proposed mass range in a single setting. The use of the well-demonstrated PRad setup allows for an essentially ready-to-run and uniquely cost-effective search for hidden sector particles in the $3 - 60$ MeV mass range with a sensitivity of 8.9$\times$10$^{-8}$ - 5.8$\times$10$^{-9}$ to $\epsilon^2$, the square of the kinetic mixing interaction constant between hidden and visible sectors. This updated proposal includes our response to the PAC49 comments.

A new electron scattering experiment (E12-21-003) to verify and understand the nature of hidden sector particles, with particular emphasis on the so-called X17 particle, has been approved at Jefferson Lab. The search for these particles is motivated by new hidden sector models introduced to account for a variety of experimental and observational puzzles: excess in $e^+e^-$ pairs observed in multiple nuclear transitions, the 4.2$\sigma$ disagreement between experiments and the standard model prediction for the muon anomalous magnetic moment, and the small-scale structure puzzle in cosmological simulations. The aforementioned X17 particle has been hypothesized to account for the excess in $e^+e^-$ pairs observed from the $^8$Be M1, $^4$He M0, and, most recently, $^{12}$C E1 nuclear transitions to their ground states observed by the ATOMKI group. This experiment will use a high resolution electromagnetic calorimeter to search for or set new limits on the production rate of the X17 and other hidden sector particles in the $3 - 60$ MeV mass range via their $e^+e^-$ decay (or $\gamma\gamma$ decay with limited tracking). In these models, the $1 - 100$ MeV mass range is particularly well-motivated and the lower part of this range still remains unexplored. Our proposed direct detection experiment will use a magnetic-spectrometer-free setup (the PRad apparatus) to detect all three final state particles in the visible decay of a hidden sector particle for an effective control of the background and will cover the proposed mass range in a single setting. The use of the well-demonstrated PRad setup allows for an essentially ready-to-run and uniquely cost-effective search for hidden sector particles in the $3 - 60$ MeV mass range with a sensitivity of 8.9$\times$10$^{-8}$ - 5.8$\times$10$^{-9}$ to $\epsilon^2$, the square of the kinetic mixing interaction constant between hidden and visible sectors.

Positron beams, both polarized and unpolarized, are identified as essential ingredients for the experimental programs at the next generation of lepton accelerators. In the context of the hadronic physics program at Jefferson Lab (JLab), positron beams are complementary, even essential, tools for a precise understanding of the electromagnetic structure of nucleons and nuclei, in both the elastic and deep-inelastic regimes. For instance, elastic scattering of polarized and unpolarized electrons and positrons from the nucleon enables a model independent determination of its electromagnetic form factors. Also, the deeply-virtual scattering of polarized and unpolarized electrons and positrons allows unambiguous separation of the different contributions to the cross section of the lepto-production of photons and of lepton-pairs, enabling an accurate determination of the nucleons and nuclei generalized parton distributions, and providing an access to the gravitational form factors. Furthermore, positron beams offer the possibility of alternative tests of the Standard Model of particle physics through the search of a dark photon, the precise measurement of electroweak couplings, and the investigation of charged lepton flavor violation. This document discusses the perspectives of an experimental program with high duty-cycle positron beams at JLab.