Incorporation of 5-fluorouracil into U2 snRNA blocks pseudouridylation and pre-mRNA splicing in vivo

5-fluorouracil (5FU) is an effective anti-cancer drug, yet its mechanism of action remains unclear. Here, we examine the effect of 5FU on pre-mRNA splicing in vivo. Using RT–PCR, we show that the splicing of a number of pre-mRNAs is inhibited in HeLa cells that have been exposed to a low dose of 5FU. It appears that this inhibitory effect is not due to its incorporation into pre-mRNA, because partially or fully 5FU-substituted pre-mRNA, when injected into Xenopus oocytes, is spliced just as well as is the unsubstituted pre-mRNA. Detailed analyses of 5FU-treated cells indicate that 5FU is incorporated into U2 snRNA at important naturally occurring pseudouridylation sites. Remarkably, 5FU incorporation effectively blocks the formation of important pseudouridines in U2 snRNA, as only a trace of pseudouridine is detected when cells are exposed to a low dose of 5FU for 5 days. Injection of the hypopseudouridylated HeLa U2 snRNA into U2-depleted Xenopus oocytes fails to reconstitute pre-mRNA splicing, whereas control U2 isolated from untreated or uracil-treated HeLa cells completely reconstitutes the splicing. Our results demonstrate for the first time that 5FU incorporates into a spliceosomal snRNA at natural pseudouridylation sites in vivo, thereby inhibiting snRNA pseudouridylation and splicing. This mechanism may contribute substantially to 5FU-mediated cell death

Hydrodynamic coefficients of oscillating flat plates at 0.15 ⩽ KC⩽ 3.15

This article presents an experimental investigation on the hydrodynamic coefficients of oscillating flat plates. The plates are forced to oscillate harmonically in still water. The range of Keulegan–Carpenter number (KC= 2 πa/ D, where a is the single amplitude of oscillation and D is the equivalent diameter of the plate) is 0.15 ⩽ KC⩽ 3.15. The hydrodynamic forces acting on the plates are measured and the hydrodynamic coefficients including added mass and damping coefficients are calculated using the Morison’s equation. The influences of the thickness ratio, shape, edge corner radius, perforation ratio and hole size on the hydrodynamic coefficients of a single plate are analyzed and presented. For the twin- and triplet-plate configurations, the spacing effects are also evaluated

Flow around an oscillating circular disk at low to moderate Reynolds numbers

Direct numerical simulations of the flow induced by a circular disk oscillating sinusoidally along its axis are performed. The aspect ratio of the disk is 10. The Reynolds number , based on the maximum speed and the diameter of the disk, is in the range of . The Keulegan-Carpenter number is in the range of . Five flow regimes are observed in the considered-space: (I) axisymmetric flow (AS), (II) planar symmetric flow in the low-region (PSL), (III) azimuthally rotating flow in the low-region (ARL), (IV) planar symmetric flow in the high-region (PSH) and (V) azimuthally rotating flow in the high-region (ARH). The critical boundaries between different flow regimes are identified based on the evolutions of the magnitude and direction of transverse force acting on the disk. For the non-axisymmetric flow regimes, the flow is one-sided with respect to the axis of the disk and is associated with a non-zero mean value of the transverse force acting on the disk

Flow around an inclined circular disk

Direct numerical simulations are performed for the uniform flow around an inclined circular disk. The diameter–thickness aspect ratio ( X=D/td) of the disk is 50 and the inclination angle (\alpha) is considered over the range of 0<6<80 deg, where \alpha=0 deg refers to the condition where the flow is normal to the disk. The Reynolds number (Re), based on the short axis of projection in the streamwise direction, is defined as Re=U_\inf D cos \alpha /\mu, where U_\inf is the velocity of the flow and \mu is the kinematic viscosity. Re is investigated over the range of 50 < Re < 300. In the considered Re–\alpha parametric space, five states are observed and denoted as: (I) steady state (SS); (II) periodic state (PS); (III) periodic state with a low frequency modulation (PSL); (IV) quasi-periodic state (QP) and (V) chaotic state (CS). Both Re and \alpha affect the bifurcation mechanism. The bifurcating sequence occurring at \alpha=0 deg is generally observed over the whole Re–\alpha space, although it is advanced at small \alpha and delayed at large \alpha. The advancement of thresholds for different states is due to the effects introduced by inclination, which tend to select the plane of symmetry for the wake in order to regulate the wake and intensify some flow features. Nevertheless, the bifurcations are still in the dominant position when leading a state without stable symmetry, i.e. the planar symmetry could not be recovered by small \alpha. These phenomena are further discussed with respect to the vortex shedding patterns behind the disk. Furthermore, for any fixed disk, the wake behaviour is only associated with that found in the steady vertical state of a freely falling disk. The fully coupled fluid–body system is fundamentally different from the fixed cases

Thiophene-Bridged Donor–Acceptor sp2-Carbon-Linked 2D Conjugated Polymers as Photocathodes for Water Reduction

Photoelectrochemical (PEC) water reduction, converting solar energy into environmentally friendly hydrogen fuel, requires delicate design and synthesis of semiconductors with appropriate bandgaps, suitable energy levels of the frontier orbitals, and high intrinsic charge mobility. In this work, the synthesis of a novel bithiophene-bridged donor–acceptor-based 2D sp2-carbon-linked conjugated polymer (2D CCP) is demonstrated. The Knoevenagel polymerization between the electron-accepting building block 2,3,8,9,14,15-hexa(4-formylphenyl) diquinoxalino[2,3-a:2′,3′-c]phenazine (HATN-6CHO) and the first electron-donating linker 2,2′-([2,2′-bithiophene]-5,5′-diyl)diacetonitrile (ThDAN) provides the 2D CCP-HATNThDAN (2D CCP-Th). Compared with the corresponding biphenyl-bridged 2D CCP-HATN-BDAN (2D CCP-BD), the bithiophene-based 2D CCP-Th exhibits a wide light-harvesting range (up to 674 nm), a optical energy gap (2.04 eV), and highest energy occupied molecular orbital–lowest unoccupied molecular orbital distributions for facilitated charge transfer, which make 2D CCP-Th a promising candidate for PEC water reduction. As a result, 2D CCP-Th presents a superb H2-evolution photocurrent density up to ≈7.9 µA cm−2 at 0 V versus reversible hydrogen electrode, which is superior to the reported 2D covalent organic frameworks and most carbon nitride materials (0.09–6.0 µA cm−2). Density functional theory calculations identify the thiophene units and cyano substituents at the vinylene linkage as active sites for the evolution of H2. © 2020 The Authors. Advanced Materials published by Wiley-VCH Gmb

Effects of Combined Simultaneous and Sequential Endostar and Cisplatin Treatment in a Mice Model of Gastric Cancer Peritoneal Metastases

Objective. Aimed to study the effects of endostar and cisplatin using an in vivo imaging system (IVIS) in a model of peritoneal metastasis of gastric cancer. Methods. NUGC-4 gastric cancer cells transfected with luciferase gene (NUGC-4-Luc) were injected i.p. into nude mice. One week later, mice were randomly injected i.p.: group 1, cisplatin (d1–3) + endostar (d4–7); group 2, endostar (d1–4) + cisplatin (d5–7); group 3, endostar + cisplatin d1, 4, and 7; group 4, saline for two weeks. One week after the final administration, mice were sacrificed. Bioluminescent data, microvessel density (MVD), and lymphatic vessel density (LVD) were analyzed. Results. Among the four groups, there were no significant differences in the weights and in the number of cancer cell photons on days 1 and 8 (P>0.05). On day 15, the numbers in groups 3 and 1 were less than that in group 2 (P<0.05). On day 21, group 3 was significantly less than group 2 (P<0.05). MVD of group 4 was less than that of groups 1 and 2 (P<0.01). There was no significant difference between groups 2 and 3 (P>0.05) or in LVD number among the four groups (P>0.05). Conclusions. IVIS® was more useful than weight, volume of ascites, and number of peritoneal nodules. The simultaneous group was superior to sequential groups in killing cancer cells and inhibiting vascular endothelium. Cisplatin-endostar was superior to endostar-cisplatin in killing cancer cells, while the latter in inhibiting peritoneal vascular endothelium

Wafer-sized multifunctional polyimine-based two-dimensional conjugated polymers with high mechanical stiffness

One of the key challenges in two-dimensional (2D) materials is to go beyond graphene, a prototype 2D polymer (2DP), and to synthesize its organic analogues with structural control at the atomic- or molecular-level. Here we show the successful preparation of porphyrin-containing monolayer and multilayer 2DPs through Schiff-base polycondensation reaction at an air-water and liquid-liquid interface, respectively. Both the monolayer and multilayer 2DPs have crystalline structures as indicated by selected area electron diffraction. The monolayer 2DP has a thickness of∼0.7 nm with a lateral size of 4-inch wafer, and it has a Young's modulus of 267±30 GPa. Notably, the monolayer 2DP functions as an active semiconducting layer in a thin film transistor, while the multilayer 2DP from cobalt-porphyrin monomer efficiently catalyses hydrogen generation from water. This work presents an advance in the synthesis of novel 2D materials for electronics and energy-related applications

Thiophene-Based Conjugated Acetylenic Polymers with Dual Active Sites for Efficient Co-Catalyst-Free Photoelectrochemical Water Reduction in Alkaline Medium

Although being attractive materials for photoelectrochemical hydrogen evolution reaction (PEC HER) under neutral or acidic conditions, conjugated polymers still show poor PEC HER performance in alkaline medium due to the lack of water dissociation sites. Herein, we demonstrate that tailoring the polymer skeleton from poly(diethynylthieno[3,2-b]thiophene) (pDET) to poly(2,6-diethynylbenzo[1,2-b:4,5-b′]dithiophene (pBDT) and poly(diethynyldithieno[3,2-b:2′,3′-d]thiophene) (pDTT) in conjugated acetylenic polymers (CAPs) introduces highly efficient active sites for water dissociation. As a result, pDTT and pBDT, grown on Cu substrate, demonstrate benchmark photocurrent densities of 170 μA cm−2 and 120 μA cm−2 (at 0.3 V vs. RHE; pH 13), which are 4.2 and 3 times higher than that of pDET, respectively. Moreover, by combining DFT calculations and electrochemical operando resonance Raman spectroscopy, we propose that the electron-enriched Cβ of the outer thiophene rings of pDTT are the water dissociation active sites, while the −C≡C− bonds function as the active sites for hydrogen evolution. © 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH Gmb