Search

BackgroundCholangiocarcinoma (CCA) is a very difficult-to-treat cancer. Chemotherapies are little effective and response to immune checkpoint inhibitors is limited. Therefore, new therapeutic strategies need to be identified.ObjectiveWe characterised the enzyme protein arginine-methyltransferase 5 (PRMT5) as a novel therapeutic target in CCA.DesignWe evaluated the expression of PRMT5, its functional partner MEP50 and methylthioadenosine phosphorylase (MTAP)—an enzyme that modulates the sensitivity of PRMT5 to pharmacological inhibitors—in human CCA tissues. PRMT5-targeting drugs, currently tested in clinical trials for other malignancies, were assessed in human CCA cell lines and organoids, as well as in two immunocompetent CCA mouse models. Transcriptomic, proteomic and functional analyses were performed to explore the underlying antitumoural mechanisms.ResultsPRMT5 and MEP50 proteins were correlatively overexpressed in most CCA tissues. MTAP was absent in 25% of intrahepatic CCA. PRMT5-targeting drugs markedly inhibited CCA cell proliferation, synergising with cisplatin and gemcitabine and hindered the growth of cholangiocarcinoma organoids. PRMT5 inhibition blunted the expression of oncogenic genes involved in chromatin remodelling and DNA repair, consistently inducing the formation of RNA loops and promoting DNA damage. Treatment with PRMT5-targeting drugs significantly restrained the growth of experimental CCA without adverse effects and concomitantly induced the recruitment of CD4 and CD8 T cells to shrinking tumourous lesions.ConclusionPRMT5 and MEP50 are frequently upregulated in human CCA, and PRMT5-targeting drugs have significant antitumoural efficacy in clinically relevant CCA models. Our findings support the evaluation of PRMT5 inhibitors in clinical trials, including their combination with cytotoxic and immune therapies.

Human tissue three-dimensional (3D) organoid cultures have the potential to reproduce in vitrothe physiological properties and cellular architecture of the organs from which they are derived. The ability of organoid cultures derived from human stomach, liver, kidney, and colon to metabolically activate three dietary carcinogens, aflatoxin B1(AFB1), aristolochic acid I (AAI), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), was investigated. In each case, the response of a target tissue (liver for AFB1; kidney for AAI; colon for PhIP) was compared with that of a nontarget tissue (gastric). After treatment cell viabilities were measured, DNA damage response (DDR) was determined by Western blotting for p-p53, p21, p-CHK2, and γ-H2AX, and DNA adduct formation was quantified by mass spectrometry. Induction of the key xenobiotic-metabolizing enzymes (XMEs) CYP1A1, CYP1A2, CYP3A4, and NQO1 was assessed by qRT-PCR. We found that organoids from different tissues can activate AAI, AFB1, and PhIP. In some cases, this metabolic potential varied between tissues and between different cultures of the same tissue. Similarly, variations in the levels of expression of XMEs were observed. At comparable levels of cytotoxicity, organoids derived from tissues that are considered targets for these carcinogens had higher levels of adduct formation than a nontarget tissue.

We present an area-efficient, asynchronous, single-input, and multiple-output Rapid Single Flux Quantum (RSFQ) pulse round-robin distribution network. We adopt the structure of a two-output toggle flip flop (TFF) where incoming pulses are temporarily stored in a SQUID loop in the form of magnetic flux quanta and then directed to outputs in a round-robin fashion. To support additional toggle outputs, we design a new circuit based on TFFs to distribute pulses in a round-robin mechanism to more than two outputs. We also elaborate on our design methodology that can support a different number of outputs while minimizing the number of JJs and power consumption. We then demonstrate a three- and four-output round-robin distribution network constructed with only 14-JJs and 18-JJs with $14.85 ~\mu W$ and 18.86- $\mu W$ power dissipation, respectively. Our four-output design has 40%–70% fewer JJ compared to a similar-functioning four-output network composed of TFFs, DFFs, splitters, and mergers. Our design also consumes 38%–70% less power and has a 16%–64% reduced delay compared to the same four-output TFF network. Finally, we demonstrate the usability of our four-output design in the context of a periodic counting network and pulse generator.

In safety-critical applications, microcontrollers have to be tested to satisfy strict quality and performance constraints. It has been demonstrated that on-chip ring oscillators can be used as speed monitors to reliably predict the performances. However, any machine-learning (ML) model is likely to be inaccurate if trained on an inadequate dataset, and labeling data for training is quite a costly process. In this article, we present a methodology based on active learning to select the best samples to be included in the training set, significantly reducing the time and cost required. Moreover, since different speed measurements are available, we designed a multilabel technique to take advantage of their correlations. Experimental results demonstrate that the approach halves the training-set size, with respect to a random-labeling, while it increases the predictive accuracy, with respect to standard single-label ML models.

Speed of sound (SoS) is a novel imaging biomarker for assessing the biomechanical characteristics of soft tissues. SoS imaging in the pulse-echo mode using conventional ultrasound (US) systems with hand-held transducers has the potential to enable new clinical uses. Recent work demonstrated that diverging waves (DWs) from a single element (SE) transmit to outperform plane-wave sequences. However, SE transmits have severely limited power and hence produce a low signal-to-noise ratio (SNR) in echo data. We herein propose Walsh–Hadamard (WH) coded and virtual-source (VS) transmit sequences for the improved SNR in SoS imaging. We additionally present an iterative method of estimating beamforming (BF) SoS in the medium, which otherwise confounds SoS reconstructions due to beamforming inaccuracies in the images used for reconstruction. Through numerical simulations, phantom experiments, and in vivo imaging data, we show that WH is not robust against motion, which is often unavoidable in clinical imaging scenarios. Our proposed VS sequence is shown to provide the highest SoS reconstruction performance, especially robust to motion artifacts. In phantom experiments, despite having a comparable SoS root-mean-square error (RMSE) of 17.5–18.0 m/s at rest, with a minor axial probe motion of $\approx 0.67$ mm/s the RMSE for SE, WH, and VS already deteriorate to 20.2, 105.4, and 19.0 m/s, respectively, showing that WH produces unacceptable results, not robust to motion. In the clinical data, the high SNR and motion resilience of VS sequences are seen to yield superior contrast compared to SE and WH sequences.

We propose a generalized architecture for the first rapid-single-flux-quantum (RSFQ) associative memory circuit. The circuit employs hyperdimensional computing (HDC), a machine learning (ML) paradigm utilizing vectors with dimensionality in the thousands to represent information. HDC designs have small memory footprints, simple computations, and simple training algorithms compared to superconducting neural network accelerators (SNNAs), making them a better option for scalable SFQ machine learning (ML) solutions. The proposed superconducting HDC (SHDC) circuit uses entirely on-chip RSFQ memory which is tightly integrated with logic, operates at 33.3 GHz, is applicable to general ML tasks, and is manufacturable at practically useful scales given current SFQ fabrication limits. Tailored to a language recognition task, SHDC consists of $\sim$2–20 M Josephson junctions (JJs) and consumes up to three times less power than an analogous CMOS HDC circuit while achieving 78–84% higher throughput. SHDC is capable of outperforming the state of the art RSFQ SNNA, SuperNPU, by 48-99% for all benchmark NN architectures tested while occupying up to 90% less area and consuming up to nine times less power. To the best of the authors' knowledge, SHDC is currently the only superconducting ML approach feasible at practically useful scales for real-world ML tasks and capable of online learning.

The testing of integrated circuits is an important topic, particularly in safety-critical applications. This is especially true for microcontrollers (MCUs) used in the automotive industry. A critical test is the performance screening in which the maximum clock frequency of the MCU is determined. For this performance screening, indirect monitors, such as ring oscillators (ROs), are used. This article presents a holistic overview of the functional path RO from the pre-silicon to the post-silicon. The implementation of such ROs is presented, as the associated advantages in terms of area consumption, leakage, and routing. In the post-silicon phase, the functional path RO frequencies are correlated with the MCU performance using machine learning approaches.

Reliable detection and tracking of surrounding objects are indispensable for comprehensive motion prediction and planning of autonomous vehicles. Due to the limitations of individual sensors, the fusion of multiple sensor modalities is required to improve the overall detection capabilities. Additionally, robust motion tracking is essential for reducing the effect of sensor noise and improving state estimation accuracy. The reliability of the autonomous vehicle software becomes even more relevant in complex, adversarial high-speed scenarios at the vehicle handling limits in autonomous racing. In this paper, we present a modular multi-modal sensor fusion and tracking method for high-speed applications. The method is based on the Extended Kalman Filter (EKF) and is capable of fusing heterogeneous detection inputs to track surrounding objects consistently. A novel delay compensation approach enables to reduce the influence of the perception software latency and to output an updated object list. It is the first fusion and tracking method validated in high-speed real-world scenarios at the Indy Autonomous Challenge 2021 and the Autonomous Challenge at CES (AC@CES) 2022, proving its robustness and computational efficiency on embedded systems. It does not require any labeled data and achieves position tracking residuals below 0.1 m.

LiDAR object detection algorithms based on neural networks for autonomous driving require large amounts of data for training, validation, and testing. As real-world data collection and labeling are time-consuming and expensive, simulation-based synthetic data generation is a viable alternative. However, using simulated data for the training of neural networks leads to a domain shift of training and testing data due to differences in scenes, scenarios, and distributions. In this work, we quantify the sim-to-real domain shift by means of LiDAR object detectors trained with a new scenario-identical real-world and simulated dataset. In addition, we answer the questions of how well the simulated data resembles the real-world data and how well object detectors trained on simulated data perform on real-world data. Further, we analyze point clouds at the target-level by comparing real-world and simulated point clouds within the 3D bounding boxes of the targets. Our experiments show that a significant sim-to-real domain shift exists even for our scenario-identical datasets. This domain shift amounts to an average precision reduction of around $14 \,\%$ for object detectors trained with simulated data. Additional experiments reveal that this domain shift can be lowered by introducing a simple noise model in simulation. We further show that a simple downsampling method to model real-world physics does not influence the performance of the object detectors.

Hepatic, pancreatic, and biliary (HPB) organoids are powerful tools for studying development, disease, and regeneration. As organoid research expands, the need for clear definitions and nomenclature describing these systems also grows. To facilitate scientific communication and consistent interpretation, we revisit the concept of an organoid and introduce an intuitive classification system and nomenclature for describing these 3D structures through the consensus of experts in the field. To promote the standardization and validation of HPB organoids, we propose guidelines for establishing, characterizing, and benchmarking future systems. Finally, we address some of the major challenges to the clinical application of organoids.

Graphic abstract:

Several new diphenylamino- and diphenylphosphanyldialkylalanes are reported, which were characterized in solution and in the solid state, assisted by in-depth bonding analysis within the DFT framework. In the case of bulky alkyl substituents on the aluminum atom, the species are stable in their monomeric form and were structurally characterized by single crystal X-ray diffraction, expanding the relatively small field of monomeric pnictogenylalanes. In the case of oligomeric diphenylpnictogenyldimethylalanes, their reactivity toward different σ-donor ligands was studied, and several examples of monomeric adducts could be structurally characterized, including the first cyclic(alkyl)(amino)carbene complexes. The reactivity of these CAAC complexes, their oligomeric precursors, and an unstabilized monomeric aminoalane toward CO2was probed, leading to different insertion products that could be characterized. Additionally, the mechanism was elucidated by DFT calculations.

Polysiloxanes are an important class of polymers for optoelectronic applications. Novel polysiloxanes with high‐refractive index (RI) based on phenanthrenyl‐substituted monomers are prepared by a polycondensation reaction starting from various substituted dialkoxysilanes as monomers. The substitution patterns on the Si atom are systematically changed to vary the properties of the linear polymers as well as the final cured material. The two monomers with polycyclic aromatic side groups 9‐phenanthrenylmethyldimethoxysilane and 9‐phenanthrenylphenyldimethoxysilane are synthesized and fully characterized, including their single crystal X‐ray structures. Linear polysiloxanes with variations in hydride, methyl, vinyl, phenyl, and phenanthrenyl side group content are prepared by acid‐ and base‐catalyzed polycondensation reactions. Both SiH and Si vinyl substituted polymers with molecular weights up to 30 kDa and adjustable RI's from 1.52 to 1.63 are obtained and the thermally cured by Pt‐catalyzed hydrosilylation reactions. Polysiloxane resins are obtained with high‐RI's, optical transmittance above 95% and thermal stabilities up to 420°C. Long‐term thermal stability tests show transmittance values above 85% even after 60 days of thermal treatment at 180°C.

AbstractIn this study, five new complexes containing deferiprone (dfp) and N,N-donor ligands [bipyridine (bpy), 1,10-phenanthroline (phen) and ethylenediamine (en)] were synthesized: [Fe(dfp)2(bpy)](PF6) (1), [Fe(dfp)2(phen)](PF6) (2), [Cu2(dfp)2(bpy)2](PF6)2(3), [Ga(dfp)2(bpy)](PF6) (4), and [Fe(dfp)2(en)](PF6) (5). Characterization of these complexes was carried out through elemental analysis and FT-IR, and single-crystal X-ray crystallography was used to determine their structures. Whilst the polyhedron has a distorted octahedral geometry in 1, 2, 4, and 5, it adopts a distorted square-pyramidal geometry in 3. Interaction of these compounds with human serum albumin (HSA) has been investigated through electronic absorption and fluorescence titration techniques. Emission quenching was performed separately for each complex at three different temperatures and thermodynamic parameters were calculated using binding constants to better understand the power of different binding forces with the HSA. Results demonstrated that compounds interact strongly with the HSA with a static quenching mechanism. Our evaluation of the cytotoxicity of complexes against the breast cancer MCF-7 cell line showed that complex 2presents a better cytotoxicity than the standard cis-Pt. Finally, using the AutoDock 4.2 program, simulations to analyze the mechanism of complex–HSA interactions and their binding mode were carried out. Results showed that the best binding mode is located in subdomain IB for 1, 2, and 4, in I/II for 3, and in IA/IIA for 5. Communicated by Ramaswamy H. Sarma

The metathesis of carbon–carbon double bonds—the ‘reshuffling’ of their constituting carbene fragments—is a tremendously important preparative tool in industry and academia. Metathesis of heavier alkene homologues is restricted to occasional unproductive examples in phosphorus chemistry and cross-metathesis to mixed heavier alkynes. We now report the thermally induced, transition-metal-free metathesis of purpose-built unsymmetrically substituted digermenes. The A2Ge=GeAB starting materials are thus converted to symmetrically substituted derivatives of the A2Ge=GeA2and ABGe=GeAB types. The use of tethered auxiliary donors (dimethylaniline groups) in substituents B ensures intramolecular donor–acceptor stabilization of the transient germylene fragments, the intermediacy of which is proven by trapping experiments. Density functional theory calculations shed light on the thermodynamic driving force of the metathesis and validate the crucial role of the tethered donor. With an analogously equipped bridged tetragermadiene precursor (A2Ge=GeB-X-BGe=GeA2), heavier acyclic diene metathesis polymerization occurs, in analogy to the widespread acyclic diene metathesis (ADMET) polymerization in the carbon case, yielding a polydigermene.

Cell Stem Cellwas launched in 2007, and this year marks its 15thanniversary. To recognize this occasion, we asked six advisory board members to reflect on inspiring discoveries reported in Cell Stem Celland how these breakthroughs connect to their vision for the future of the field.

Pentaisopropylcyclopentadienyl dichloropnictogen complexes have been synthesized and studied in solution and in the solid state. The coordination mode of the group 15 element to the Cp ligand varies, with the lighter elements phosphorus and arsenic featuring η1σ-bonded Cp groups and the heavier elements antimony and bismuth exhibiting η3and η5bonded Cp groups, respectively. In the case of ≤η3bonded Cp groups, a rapid sigmatropic rearrangement is observed at room temperature in solution and was examined by VT-NMR spectroscopy and DFT calculations. Furthermore, bis(cyclopentadienyl)diphosphenes with tetra- and pentaisopropylcyclopentadienyl groups could be obtained in good yields from the corresponding dichlorophosphanes, and a related silver(I) complex of a diphosphene was characterized.

Liver regeneration is a complex process involving the crosstalk of multiple cell types, including hepatocytes, hepatic stellate cells, endothelial cells and inflammatory cells. The healthy liver is mitotically quiescent, but following toxic damage or resection the cells can rapidly enter the cell cycle to restore liver mass and function. During this process of regeneration, epithelial and non-parenchymal cells respond in a tightly coordinated fashion. Recent studies have described the interaction between inflammatory cells and a number of other cell types in the liver. In particular, macrophages can support biliary regeneration, contribute to fibrosis remodelling by repressing hepatic stellate cell activation and improve liver regeneration by scavenging dead or dying cells in situ. In this Review, we describe the mechanisms of tissue repair following damage, highlighting the close relationship between inflammation and liver regeneration, and discuss how recent findings can help design novel therapeutic approaches.

A detailed experimental and computational study of a series of stannocene and plumbocene N-heterocyclic carbene complexes is presented. This unique class of group 14 Lewis acid–base adducts was obtained from reactions of the corresponding metallocenes and N-heterocyclic carbenes (NHC) and was structurally characterized by single-crystal X-ray diffraction. The obtained structures show a perpendicular pose of the NHC with respect to the metallocene, hence precluding the maximum interaction between the moieties. The nature of the Sn–CNHCand Pb–CNHCbonds has been investigated by applying natural bond orbital (NBO) analysis and energy decomposition analysis (EDA-NOCV). For the sake of comparison, known stannocene and plumbocene Lewis base complexes have been included in the series. The attractive chemical bonding interactions are around 50% electrostatic, 30% covalent, and 20% dispersion. Indeed, dispersion interactions play a determining role the larger the substituents become. The covalent interactions derive from the donation of the carbene ligand into the empty p orbital of the metallocene.

The heterocyclic aromatic amine (HAA) 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is converted into 7-hydroxy-5-methyl-3-phenyl-6,7,8,9-tetrahydropyrido[3′,2′:4,5]imidazo[1,2-a]pyrimidin-5-ium chloride (PhIP-M1) via a chemical reaction with 3-hydroxypropionaldehyde or acrolein derived from glycerol by reuterin producing gut bacteria. Because it is unknown whether this reaction also applies to other HAAs, seven foodborne HAAs (2-amino-9H-pyrido[2,3-b]indole (AαC), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethyl-3H-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethyl-3H-imidazo[4,5-f]quinoxaline (MeIQx), 9H-pyrido[3,4-b]indole (norharman), and 1-methyl-9H-pyrido[3,4-b]indole (harman)) were anaerobically incubated with Lactobacillus reuteriDSM 20016 in the presence of glycerol. The extent of conversion, as analyzed by HPLC-DAD/FLD, was dependent on both the studied HAAs and the glucose/glycerol ratio, indicating reuterin to be involved in HAA metabolism. Based on HRMS analyses, PhIP-M1-type metabolites were detected for AαC, Trp-P-1, IQ, MeIQ, MeIQx, harman, and norharman. In the case of AαC, this was confirmed by metabolite isolation (AαC-M8, 2,3,4,10-tetrahydro-1H-indolo[2,3-b][1,8]naphthyridin-2-ol) and one- (1H) and two-dimensional (HSQC, HMBC, COSY, DOSY) NMR spectroscopy. In addition, based on HRMS and/or NMR spectroscopy, a new type of HAA metabolite, resulting from the reaction with two molecules of 3-hydroxypropionaldehyde or acrolein, is hypothesized for AαC, Trp-P-1, IQ, MeIQ, and MeIQx.

Proof of concept for the protection of the nucleophilic functionality of disilenidesdisila analogues of vinyllithiumwith preservation of the SiSi bond is reported. 1-Iodo-2,4,6-trimethoxybenzene (TMOP-I) reacts with lithium tris(2,4,6-triisopropylphenyl)disilenide (1), affording the disilene Tip2SiSi(Tip)TMOP (2) in high yield. The presence of the TMOP group in disilene 2enables the regioselective addition of polar substrates to the SiSi double bond, including water, ammonia, acetylenes, and isocyanides. NMR spectroscopic analysis of the reductive cleavage of the TMOP group and subsequent trapping of the corresponding disilenides with Me3SiCl reveals KC8as a highly appropriate reducing agent for the selective deprotection.

Unsaturated silicon clusters (siliconoids) are short-lived intermediates during the transition from molecules to the elemental bulk; stable representatives reiterate surface features of silicon materials. The incorporation of suitable heteroatoms into the cluster scaffold of stable siliconoids extends this analogy to the technological process of silicon doping. Here, we report boron- and phosphorus-containing heterosiliconoids with BSi5and PSi5core based on the global minimum Si6R6platform (dubbed benzpolarene for its relationship to benzene). The reductive cleavage of an SiR2moiety (R = 2,4,6-iPr3C6H2) from Si6R6selectively yields a dianionic Si5R42–cluster as its lithium salt. Treatment with Me3SiCl affords the corresponding trimethylsilyl-substituted (Me3Si)2Si5R4. Reaction of Si5R42–with iPr2NECl2(E = B, P) yields the unprecedented p- and n-doped heterosiliconoids iPr2NESi5R4. Their peculiar electronic features are compared to those of the hexasilabenzpolarene starting material on grounds of NMR spectroscopy, X-ray diffraction, and DFT calculations.

Following severe or chronic liver injury, adult ductal cells (cholangiocytes) contribute to regeneration by restoring both hepatocytes and cholangiocytes. We recently showed that ductal cells clonally expand as self-renewing liver organoids that retain their differentiation capacity into both hepatocytes and ductal cells. However, the molecular mechanisms by which adult ductal-committed cells acquire cellular plasticity, initiate organoids and regenerate the damaged tissue remain largely unknown. Here, we describe that ductal cells undergo a transient, genome-wide, remodelling of their transcriptome and epigenome during organoid initiation and in vivo following tissue damage. TET1-mediated hydroxymethylation licences differentiated ductal cells to initiate organoids and activate the regenerative programme through the transcriptional regulation of stem-cell genes and regenerative pathways including the YAP–Hippo signalling. Our results argue in favour of the remodelling of genomic methylome/hydroxymethylome landscapes as a general mechanism by which differentiated cells exit a committed state in response to tissue damage.

To explore the chemical diversity of metabolites from new species of Dothideomycetes, the ex-type strain of Sparticola junciwas investigated. Seven highly oxygenated and functionalized spirodioxynaphthalene natural products incorporating carboxyalkylidene–cyclopentanoid (1–4), carboxyl-functionalized oxabicyclo[3.3.0]octane (5–6), and annelated 2-cyclopentenone/δ-lactone (7) units, sparticolins A–G, were isolated from submerged cultures of the fungus. Their chemical structures including their relative (and absolute) configurations were established through spectroscopic and X-ray crystallographic analyses. Sparticolin B (2) exhibited inhibitory activity against the Gram-positive bacteria Bacillus subtilis, Micrococcus luteus, and Staphylococcus aureus, while sparticolin G (7) showed antifungal activities against Schizosaccharomyces pombeand Mucor hiemalis. All other sparticolins were only weakly active against S. aureusand also showed weak activities against the nematode Caenorhabditis elegans. Compounds 2and 7also showed moderate cytotoxic activities against seven mammalian cell lines.

We report the reversible coordination of the N-heterocyclic carbene (NHC), NHCiPr2Me2(NHCiPr2Me2= 1,3-diisopropyl-4,5-dimethylimidazol-2-ylidene), to silicon(IV)-halides, SiCl4, MeSiCl3, Me2SiCl2, and Me3SiCl. Predicted as well as experimentally determined thermodynamic parameters of these equilibria confirm that the complexation constant increases with the Lewis acidity of the silicon halides. In contrast, the more σ-donating N-heterocyclic carbene, NHCMe4(NHCMe4= 1,3,4,5-tetramethylimidazol-2-ylidene), does not show any signs of dissociation from the corresponding SiCl4and Me2SiCl2adducts even at higher temperatures. As a consequence, NHCiPr2Me2in donor–acceptor stabilized Si(II)- and Ge(II)-dimethyl complexes, NHCiPr2Me2·GeMe2·Fe(CO)4and NHCiPr2Me2·SiMe2·Fe(CO)4, is readily replaced by NHCMe4.