Catalytic synthesis of phenols with nitrous oxide.

The development of catalytic chemical processes that enable the revalorization of nitrous oxide (N2O) is an attractive strategy to alleviate the environmental threat posed by its emissions1-6. Traditionally, N2O has been considered an inert molecule, intractable for organic chemists as an oxidant or O-atom transfer reagent, owing to the harsh conditions required for its activation (>150 °C, 50‒200 bar)7-11. Here we report an insertion of N2O into a Ni‒C bond under mild conditions (room temperature, 1.5-2 bar N2O), thus delivering valuable phenols and releasing benign N2. This fundamentally distinct organometallic C‒O bond-forming step differs from the current strategies based on reductive elimination and enables an alternative catalytic approach for the conversion of aryl halides to phenols. The process was rendered catalytic by means of a bipyridine-based ligands for the Ni centre. The method is robust, mild and highly selective, able to accommodate base-sensitive functionalities as well as permitting phenol synthesis from densely functionalized aryl halides. Although this protocol does not provide a solution to the mitigation of N2O emissions, it represents a reactivity blueprint for the mild revalorization of abundant N2O as an O source.

K63-Ubiquitylation and TRAF6 Pathways Regulate Mammalian P-Body Formation and mRNA Decapping.

Signals and posttranslational modifications regulating the decapping step in mRNA degradation pathways are poorly defined. In this study we reveal the importance of K63-linked ubiquitylation for the assembly of decapping factors, P-body formation, and constitutive decay of instable mRNAs encoding mediators of inflammation by various experimental approaches. K63-branched ubiquitin chains also regulate IL-1-inducible phosphorylation of the P-body component DCP1a. The E3 ligase TRAF6 binds to DCP1a and indirectly regulates DCP1a phosphorylation, expression of decapping factors, and gene-specific mRNA decay. Mutation of six C-terminal lysines of DCP1a suppresses decapping activity and impairs the interaction with the mRNA decay factors DCP2, EDC4, and XRN1, but not EDC3, thus remodeling P-body architecture. The usage of ubiquitin chains for the proper assembly and function of the decay-competent mammalian decapping complex suggests an additional layer of control to allow a coordinated function of decapping activities and mRNA metabolism in higher eukaryotes.

Bismuth-Catalyzed Amide Reduction.

In this article we report that a cationic version of Akiba's BiIII complex catalyzes the reduction of amides to amines using silane as hydride donor. The catalytic system features low catalyst loadings and mild conditions, en route to secondary and tertiary aryl- and alkylamines. The system tolerates functional groups such as alkene, ester, nitrile, furan and thiophene. Kinetic studies on the reaction mechanism result in the identification of a reaction network with an important product inhibition that is in agreement with the experimental reaction profiles.

Radical Activation of N-H and O-H Bonds at Bismuth(II).

The development of unconventional strategies for the activation of ammonia (NH3) and water (H2O) is of capital importance for the advancement of sustainable chemical strategies. Herein we provide the synthesis and characterization of a radical equilibrium complex based on bismuth featuring an extremely weak Bi-O bond, which permits the in situ generation of reactive Bi(II) species. The ensuing organobismuth(II) engages with various amines and alcohols and exerts an unprecedented effect onto the X-H bond, leading to low BDFEX-H. As a result, radical activation of various N-H and O-H bonds─including ammonia and water─occurs in seconds at room temperature, delivering well-defined Bi(III)-amido and -alkoxy complexes. Moreover, we demonstrate that the resulting Bi(III)-N complexes engage in a unique reactivity pattern with the triad of H+, H-, and H• sources, thus providing alternative pathways for main group chemistry.

Identification of compounds that bind the centriolar protein SAS-6 and inhibit its oligomerization.

Centrioles are key eukaryotic organelles that are responsible for the formation of cilia and flagella, and for organizing the microtubule network and the mitotic spindle in animals. Centriole assembly requires oligomerization of the essential protein spindle assembly abnormal 6 (SAS-6), which forms a structural scaffold templating the organization of further organelle components. A dimerization interaction between SAS-6 N-terminal "head" domains was previously shown to be essential for protein oligomerization in vitro and for function in centriole assembly. Here, we developed a pharmacophore model allowing us to assemble a library of low-molecular-weight ligands predicted to bind the SAS-6 head domain and inhibit protein oligomerization. We demonstrate using NMR spectroscopy that a ligand from this family binds at the head domain dimerization site of algae, nematode, and human SAS-6 variants, but also that another ligand specifically recognizes human SAS-6. Atomistic molecular dynamics simulations starting from SAS-6 head domain crystallographic structures, including that of the human head domain which we now resolve, suggest that ligand specificity derives from favorable Van der Waals interactions with a hydrophobic cavity at the dimerization site.

Diversity and distribution of the bmp gene cluster and its Polybrominated products in the genus Pseudoalteromonas.

The production of pentabromopseudilin and related brominated compounds by Pseudoalteromonas spp. has recently been linked to the bmp biosynthetic gene cluster. This study explored the distribution and evolutionary history of this gene cluster in the genus Pseudoalteromonas. A phylogeny of the genus revealed numerous clades that do not contain type strains, suggesting considerable species level diversity has yet to be described. Comparative genomics revealed four distinct versions of the gene cluster distributed among 19 of the 101 Pseudoalteromonas genomes examined. These were largely localized to the least inclusive clades containing the Pseudoalteromonas luteoviolacea and Pseudoalteromonas phenolica type strains and show clear evidence of gene and gene cluster loss in certain lineages. Bmp gene phylogeny is largely congruent with the Pseudoalteromonas species phylogeny, suggesting vertical inheritance within the genus. However, the gene cluster is found in three different genomic environments suggesting either chromosomal rearrangement or multiple acquisition events. Bmp conservation within certain lineages suggests the encoded products are highly relevant to the ecology of these bacteria.

Left ventricular blood flow patterns at rest and under dobutamine stress in healthy pigs.

Intracardiac blood flow patterns are affected by the morphology of cardiac structures and are set up to support the heart's pump function. Exercise affects contractility and chamber size as well as pre- and afterload. The aim of this study was to test the feasibility of four-dimensional phase contrast cardiovascular MRI under pharmacological stress and to study left ventricular blood flow under stress. 4D flow data were successfully acquired and analysed in 12 animals. During dobutamine infusion, heart rate and ejection fraction increased (82 ± 5 bpm versus 124 ± 3 bpm/46 ± 9% versus 65 ± 7%; both p < 0.05). A decrease in left ventricular end-diastolic volume (72 ± 14 mL versus 55 ± 8 mL; p < 0.05) and end-systolic volume (40 ± 15 mL versus 19 ± 6 mL; p < 0.05) but no change in stroke volume were observed. Trans-mitral diastolic inflow velocity increased under dobutamine and the trajectory of inflowing blood was directed towards the anterior septum with increased inflow angle (26 ± 5°) when compared with controls (15 ± 2°). In 5/6 animals undergoing stress diastolic vortices developed later, and in 3/6 animals vortices collapsed earlier with significantly smaller cross-sectional area during diastole. The vorticity index was not affected. Under the stress condition direct flow (% ejection within the next heart beat) increased from 43 ± 6% to 53 ± 8%. 4D MRI blood flow acquisition and analysis are feasible in pig hearts under dobutamine-induced stress. Flow patterns characterized by high blood velocity and antero-septally oriented diastolic inflow as well as decreased ventricular volumes are unfavourable conditions for diastolic vortex development under pharmacological stress, and cardiac output is increased by a rise in heart rate and directly ejected left ventricular blood volume.

Metagenomic discovery of polybrominated diphenyl ether biosynthesis by marine sponges.

Naturally produced polybrominated diphenyl ethers (PBDEs) pervade the marine environment and structurally resemble toxic man-made brominated flame retardants. PBDEs bioaccumulate in marine animals and are likely transferred to the human food chain. However, the biogenic basis for PBDE production in one of their most prolific sources, marine sponges of the order Dysideidae, remains unidentified. Here, we report the discovery of PBDE biosynthetic gene clusters within sponge-microbiome-associated cyanobacterial endosymbionts through the use of an unbiased metagenome-mining approach. Using expression of PBDE biosynthetic genes in heterologous cyanobacterial hosts, we correlate the structural diversity of naturally produced PBDEs to modifications within PBDE biosynthetic gene clusters in multiple sponge holobionts. Our results establish the genetic and molecular foundation for the production of PBDEs in one of the most abundant natural sources of these molecules, further setting the stage for a metagenomic-based inventory of other PBDE sources in the marine environment.

Quantitative myocardial first-pass cardiovascular magnetic resonance perfusion imaging using hyperpolarized [1-13C] pyruvate.

BACKGROUND: The feasibility of absolute myocardial blood flow quantification and suitability of hyperpolarized [1-13C] pyruvate as contrast agent for first-pass cardiovascular magnetic resonance (CMR) perfusion measurements are investigated with simulations and demonstrated in vivo in a swine model. METHODS: A versatile simulation framework for hyperpolarized CMR subject to physical, physiological and technical constraints was developed and applied to investigate experimental conditions for accurate perfusion CMR with hyperpolarized [1-13C] pyruvate. Absolute and semi-quantitative perfusion indices were analyzed with respect to experimental parameter variations and different signal-to-noise ratio (SNR) levels. Absolute myocardial blood flow quantification was implemented with an iterative deconvolution approach based on Fermi functions. To demonstrate in vivo feasibility, velocity-selective excitation with an echo-planar imaging readout was used to acquire dynamic myocardial stress perfusion images in four healthy swine. Arterial input functions were extracted from an additional image slice with conventional excitation that was acquired within the same heartbeat. RESULTS: Simulations suggest that obtainable SNR and B0 inhomogeneity in vivo are sufficient for the determination of absolute and semi-quantitative perfusion with ≤25% error. It is shown that for expected metabolic conversion rates, metabolic conversion of pyruvate can be neglected over the short duration of acquisition in first-pass perfusion CMR. In vivo measurements suggest that absolute myocardial blood flow quantification using hyperpolarized [1-13C] pyruvate is feasible with an intra-myocardial variability comparable to semi-quantitative perfusion indices. CONCLUSION: The feasibility of quantitative hyperpolarized first-pass perfusion CMR using [1-13C] pyruvate has been investigated in simulations and demonstrated in swine. Using an approved and metabolically active compound is envisioned to increase the value of hyperpolarized perfusion CMR in patients.

A multisample dissolution dynamic nuclear polarization system for serial injections in small animals.

PURPOSE: Several in vivo applications of dissolution dynamic nuclear polarization (DNP) require rapid successive injections of hyperpolarized substrates. Here we present the design and performance of a custom-built multisample dissolution DNP setup for small animal research. METHODS: The DNP setup consists of a commercial wide-bore magnet charged to 3.35 T, a cryostat, a 94-GHz microwave source, and a custom-built skeleton that accommodates four identical sample sticks. Each sample stick features a dissolver locked into the skeleton port and a lifter, which permits moving the sample cup out of the liquid helium bath for dissolution. RESULTS: The dissolution of the first sample was triggered after 2 hours of polarization buildup during single-shot operation of the cryostat. Thereafter, a time window of 75-90 min was available to dissolve the remaining three polarized samples. The average liquid state polarization over all four sticks was measured as 18.7% ± 2.3% for [1-13C] pyruvate 30 s after dissolution. In vivo applicability of the setup using serial injections of [1-13C] pyruvate to study cardiac metabolism in rats revealed good reproducibility. CONCLUSION: The proposed four-sample DNP insert provides reproducible liquid state polarization of [1-13C] pyruvate and allows for rapid repeat injections in small animals. Magn Reson Med 77:904-910, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Image-based background phase error correction in 4D flow MRI revisited.

PURPOSE: To correct background phase errors in phase-contrast magnetic resonance imaging (MRI), image-based correction by referencing through stationary tissue is widely used. The aim of the present study was a detailed assessment of background phase errors in 4D Flow MRI and limitations of image-based correction. MATERIALS AND METHODS: In a phantom study, 4D Flow MRI data were acquired for typical settings on two clinical 3T MR systems. Background errors were analyzed with respect to their spatial order and minimum requirements regarding the signal-to-noise ratio (SNR) and the amount of stationary tissue for image-based correction were assessed. For in vivo evaluation, data of the aorta were acquired on one 3T MR system in five healthy subjects including subsequent scans on the stationary phantom as reference. RESULTS: Background errors were found to exhibit spatial variation of first- to third-order. For correction, a minimum SNR of 20 was needed to achieve an error of less than 0.4% of the encoding velocity. The minimum amount of stationary tissue was strongly dependent on the spatial order requiring at least 25%, 60%, and 75% of stationary tissue for first-, second-, and third-order correction. In vivo evaluation showed that with 35-41% of stationary tissue available only first-order correction yielded a significant reduction (P < 0.01). CONCLUSION: Background phase errors can range from first to third spatial order in 4D Flow MRI requiring correction with appropriate polynomials. At the same time, the limited amount of stationary tissue available in vivo limits image-based background phase correction to first spatial order. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2017;46:1516-1525.

Hyperpolarized 13C urea myocardial first-pass perfusion imaging using velocity-selective excitation.

BACKGROUND: A velocity-selective binomial excitation scheme for myocardial first-pass perfusion measurements with hyperpolarized 13C substrates, which preserves bolus magnetization inside the blood pool, is presented. The proposed method is evaluated against gadolinium-enhanced 1H measurements in-vivo. METHODS: The proposed excitation with an echo-planar imaging readout was implemented on a clinical CMR system. Dynamic myocardial stress perfusion images were acquired in six healthy pigs after bolus injection of hyperpolarized 13C urea with the velocity-selective vs. conventional excitation, as well as standard 1H gadolinium-enhanced images. Signal-to-noise, contrast-to-noise (CNR) and homogeneity of semi-quantitative perfusion measures were compared between methods based on first-pass signal-intensity time curves extracted from a mid-ventricular slice. Diagnostic feasibility is demonstrated in a case of septal infarction. RESULTS: Velocity-selective excitation provides over three-fold reduction in blood pool signal with a two-fold increase in myocardial CNR. Extracted first-pass perfusion curves reveal a significantly reduced variability of semi-quantitative first-pass perfusion measures (12-20%) for velocity-selective excitation compared to conventional excitation (28-93%), comparable to that of reference 1H gadolinium data (9-15%). Overall image quality appears comparable between the velocity-selective hyperpolarized and gadolinium-enhanced imaging. CONCLUSION: The feasibility of hyperpolarized 13C first-pass perfusion CMR has been demonstrated in swine. Comparison with reference 1H gadolinium data revealed sufficient data quality and indicates the potential of hyperpolarized perfusion imaging for human applications.

LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean).

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the detection and quantitation of karlotoxins in the selected reaction monitoring (SRM) mode. This novel method was based upon the analysis of purified karlotoxins (KcTx-1, KmTx-2, 44-oxo-KmTx-2, KmTx-5), one amphidinol (AM-18), and unpurified extracts of bulk cultures of the marine dinoflagellate Karlodinium veneficum strain CCMP2936 from Delaware (Eastern USA), which produces KmTx-1 and KmTx-3. The limit of detection of the SRM method for KmTx-2 was determined as 2.5 ng on-column. Collision induced dissociation (CID) spectra of all putative karlotoxins were recorded to present fragmentation patterns of each compound for their unambiguous identification. Bulk cultures of K. veneficum strain K10 isolated from an embayment of the Ebro Delta, NW Mediterranean, yielded five previously unreported putative karlotoxins with molecular masses 1280, 1298, 1332, 1356, and 1400 Da, and similar fragments to KmTx-5. Analysis of several isolates of K. veneficum from the Ebro Delta revealed small-scale diversity in the karlotoxin spectrum in that one isolate from Fangar Bay produced KmTx-5, whereas the five putative novel karlotoxins were found among several isolates from nearby, but hydrographically distinct Alfacs Bay. Application of this LC-MS/MS method represents an incremental advance in the determination of putative karlotoxins, particularly in the absence of a complete spectrum of purified analytical standards of known specific potency.

SmartFluo: A Method and Affordable Adapter to Measure Chlorophyll a Fluorescence with Smartphones.

In order to increase the monitoring capabilities of inland and coastal waters, there is a need for new, affordable, sensitive and mobile instruments that could be operated semi-automatically in the field. This paper presents a prototype device to measure chlorophyll a fluorescence: the SmartFluo. The device is a combination of a smartphone offering an intuitive operation interface and an adapter implying a cuvette holder, as well as a suitable illumination source. SmartFluo is based on stimulated fluorescence of water constituents such as chlorophyll a. The red band of the digital smartphone camera is sensitive enough to detect quantitatively the characteristic red fluorescence emission. The adapter contains a light source, a strong light emitting diode and additional filters to enhance the signal-to-noise ratio and to suppress the impact of scattering. A novel algorithm utilizing the red band of the camera is provided. Laboratory experiments of the SmartFluo show a linear correlation (R 2 = 0.98) to the chlorophyll a concentrations measured by reference instruments, such as a high-performance benchtop laboratory fluorometer (LS 55, PerkinElmer).

Hyperpolarized Metabolic MR Imaging of Acute Myocardial Changes and Recovery after Ischemia-Reperfusion in a Small-Animal Model.

PURPOSE: To implement hyperpolarized magnetic resonance (MR) imaging in an animal model of ischemia-reperfusion and to assess in vivo the regional changes in pyruvate metabolism within the 1st hour and at 1 week after a brief episode of coronary occlusion and reperfusion. MATERIALS AND METHODS: All animal experiments were performed with adherence to the Swiss Animal Protection law and were approved by the regional veterinary office. A closed-chest rat model was implemented by using an inflatable balloon secured around the left coronary artery. Animals were placed in an MR system 5-7 days after surgery. [1-(13)C]pyruvate was polarized by using a home-built multisample hyperpolarizer. Hyperpolarized pyruvate was injected at five stages: at baseline; at reperfusion after 15 minutes of coronary occlusion; and at 30 minutes, 60 minutes, and 1 week after ischemia reperfusion. The conversion of pyruvate into lactate and bicarbonate was imaged by using dedicated MR sequences alongside wall motion and delayed enhancement imaging. After imaging, the heart was removed and stained to delineate the area at risk (AAR). Differences between AAR and remote myocardium were assessed by using a repeated measures analysis of variance and a post hoc Bonferroni multiple comparison test. RESULTS: Data were collected in 12 animals. Occlusion led to hypokinesia of the anterior or anterolateral segments of the myocardium. At reperfusion, the average lactate-to-bicarbonate ratio increased in the AAR relative to that at baseline (from 1.93 ± 0.48 to 3.01 ± 0.74, P < .001) and was significantly higher when compared with that in the remote area (1.91 ± 0.38, P < .001). In the 60 minutes after occlusion, the lactate-to-bicarbonate ratio in the AAR recovered but was still elevated relative to that in the remote area. One week after ischemia-reperfusion, no difference between AAR and remote area could be detected. CONCLUSION: Hyperpolarized metabolic MR imaging can be used to successfully detect acute changes in [1-(13)C]pyruvate metabolism after ischemia-reperfusion, thereby enabling in vivo monitoring of the metabolic effects of reperfusion strategies.

Accelerating 4D flow MRI by exploiting vector field divergence regularization.

PURPOSE: To improve velocity vector field reconstruction from undersampled four-dimensional (4D) flow MRI by penalizing divergence of the measured flow field. THEORY AND METHODS: Iterative image reconstruction in which magnitude and phase are regularized separately in alternating iterations was implemented. The approach allows incorporating prior knowledge of the flow field being imaged. In the present work, velocity data were regularized to reduce divergence, using either divergence-free wavelets (DFW) or a finite difference (FD) method using the ℓ1-norm of divergence and curl. The reconstruction methods were tested on a numerical phantom and in vivo data. Results of the DFW and FD approaches were compared with data obtained with standard compressed sensing (CS) reconstruction. RESULTS: Relative to standard CS, directional errors of vector fields and divergence were reduced by 55-60% and 38-48% for three- and six-fold undersampled data with the DFW and FD methods. Velocity vector displays of the numerical phantom and in vivo data were found to be improved upon DFW or FD reconstruction. CONCLUSION: Regularization of vector field divergence in image reconstruction from undersampled 4D flow data is a valuable approach to improve reconstruction accuracy of velocity vector fields.

Image reconstruction using a gradient impulse response model for trajectory prediction.

PURPOSE: Gradient imperfections remain a challenge in MRI, especially for sequences relying on long imaging readouts. This work aims to explore image reconstruction based on k-space trajectories predicted by an impulse response model of the gradient system. THEORY AND METHODS: Gradient characterization was performed twice with 3 years interval on a commercial 3 Tesla (T) system. The measured gradient impulse response functions were used to predict actual k-space trajectories for single-shot echo-planar imaging (EPI), spiral and variable-speed EPI sequences. Image reconstruction based on the predicted trajectories was performed for phantom and in vivo data. Resulting images were compared with reconstructions based on concurrent field monitoring, separate trajectory measurements, and nominal trajectories. RESULTS: Image reconstruction using model-based trajectories yielded high-quality images, comparable to using separate trajectory measurements. Compared with using nominal trajectories, it strongly reduced ghosting, blurring, and geometric distortion. Equivalent image quality was obtained with the recent characterization and that performed 3 years prior. CONCLUSION: Model-based trajectory prediction enables high-quality image reconstruction for technically challenging sequences such as single-shot EPI and spiral imaging. It thus holds great promise for fast structural imaging and advanced neuroimaging techniques, including functional MRI, diffusion tensor imaging, and arterial spin labeling. The method can be based on a one-time system characterization as demonstrated by successful use of 3-year-old calibration data. Magn Reson Med 76:45-58, 2016. © 2015 Wiley Periodicals, Inc.

Hybrid multiband excitation multiecho acquisition for hyperpolarized (13) C spectroscopic imaging.

PURPOSE: Fast dynamic imaging of hyperpolarized (13) C-labeled pyruvate and its downstream metabolites shows great potential for probing metabolic changes in the heart. Sequences that allow for fast encoding of the spectral and spatial information of the myocardial metabolism and optimal signal excitation are usually limited by gradient performance, especially at high magnetic fields. Here we propose a combination of a spectral-spatial multiband excitation and multiecho readout to overcome these limitations. METHODS: By using a low-bandwidth, two-pulse excitation, a thinner slice compared with conventional spectral-spatial excitation is achieved, while at the same time allowing for low flip angle excitation on pyruvate and high flip angle excitation on bicarbonate and lactate, which optimizes signal-to-noise ratio (SNR) in cardiac metabolic imaging. The implementation was evaluated in 13 healthy female Sprague-Dawley rats at 9.4T. RESULTS: Using a slice thickness of 4 mm, a mean (± standard deviation) peak SNR of 18.3 (±8.4), 15.2 (±6.6), and 8.6 (±2.0) was observed for pyruvate, lactate, and bicarbonate, respectively. CONCLUSION: This approach provides high SNR in metabolic images while at the same time allowing for a thin slice selection even at high magnetic fields. This is crucial in metabolic imaging in small animal models.