Clinical evaluation of high-resolution cone-beam computed tomography for the implantation of flow-diverter stents in intracranial aneurysms.

It is well-established that complete expansion and proper apposition to the vessel wall of flow-diverter stents are critical for optimizing endovascular aneurysm outcomes by using flow diversion techniques. We aimed to evaluate the clinical value of high-resolution cone-beam CT (HR-CBCT) upon flow-diverter stent implantation in intracranial aneurysms. In this study, we retrospectively analyzed the clinical data of eighty-one patients (101 intracranial aneurysms) who underwent flow-diverter stent implantation (Pipeline™ or Tubridge™). Images were reconstructed using conventional cone-beam CT (CBCT)(voxel size 0.43 mm isotropic) and HR-CBCT(voxel size 0.15 mm isotropic). Immediately after stent deployment, dual volume 3D fusion images were obtained from 3D-digital subtraction angiography (DSA) and HR-CBCT. The image quality for stent visualization was graded from 0 to 2 (0:not able to assess, 1:limited, but able to assess; 2:clear visualization), and the stent expansion status (full, under-expanded or poor apposition) was also recorded. Finally, patients were treated using flow-diverter stents (n = 92: 17 Pipeline and 75 Tubridge). Compared to CBCT, HR-CBCT led to improved visualization of the structures of the stents and significantly improved the image quality (mean score: 0.59 ± 0.67 vs. 1.6 ± 0.63, P < 0.001). For 28 stents (seven Pipeline and 21 Tubridge), partially incomplete apposition was observed by HR-CBCT but not by conventional CBCT and resolved by microguidewire looping dilation or balloon dilation. High-resolution cone-beam CT could better display flow-diverter stent details and yielded an improved image quality, which facilitated the assessment of stent deployment, potentially reducing the incidence of complications.

Clinical Study of High-Resolution C-Arm CT in Mechanical Recanalization and Stent Implantation for Chronic Cerebral Artery Occlusion.

Background: In recent years, among patients with chronic cerebral artery occlusion, recanalization can be achieved by an endovascular operation. However, complications and restenosis rates remain high. Objective: To evaluate the utility of high-resolution C-arm CT (Dyna micro-CT) for stent placement in patients with chronic cerebral artery occlusion. Methods and Materials: We retrospectively reviewed the clinical data of 27 patients with chronic cerebral artery occlusion who underwent mechanical recanalization and stent implantation. Images were reconstructed using conventional C-arm CT (Dyna CT) and Dyna micro-CT. Whether the stent was fully expanded and image quality was evaluated. Follow-up assessments included clinical and angiographic outcomes and complications. Results: Twenty-two patients successfully underwent stenting (22 stents; 14 cases: Neuroform EZ; eight cases: Enterprise); stenting failed in five patients. Compared to Dyna CT, Dyna micro-CT afforded improved visualization of the stent structure, providing significantly improved image quality (P < 0.05). In seven patients, the stent under-expanded and dilatation was performed; thereafter, stent malapposition improved. One patient experienced sudden headache 22 hours after the procedure; CT showed intraparenchymal hemorrhage. The remaining 21 patients did not have acute thrombosis or bleeding complications and were followed up by imaging for 3-6 months. In three patients, digital subtraction angiography showed mild in-stent stenosis. Conclusions: High-resolution C-arm CT can improve visualization of stent structures in chronic cerebral artery occlusion, making it easy to determine the extent of stent deployment and potentially reduce complications and stent restenosis.

The actions of neonicotinoid insecticides on nicotinic acetylcholine subunits Ldα1 and Ldα8 of Leptinotarsa decemlineata and assembled receptors.

The insect nicotinic acetylcholine receptor (nAChR) is a pentameric channel protein and also a target for neonicotinoids. There are few reported studies on the molecular interactions of Leptinotarsa decemlineata nAChRs with neonicotinoids. In this study, we analyzed the response of acetylcholine and neonicotinoids (thiamethoxam [TMX], imidacloprid [IMI], and clothianidin [CLO]) on hybrid receptors constructed by nAChR α1 and α8 subunits of L. decemlineata (Ldα1 and Ldα8) co-expressed with rat ß2 subunit (rß2) at different capped RNA (cRNA) ratios in Xenopus oocytes. In addition, we evaluated the expression changes of Ldα1 and Ldα8 after median lethal dose of TMX treatment for 72 h by quantitative polymerase chain reaction (qPCR). The resulting functional nAChRs Ldα1/rß2 and Ldα1/Ldα8/rß2 showed different pharmacological characteristics. The neonicotinoids tested showed lower agonist affinity on Ldα1/Ldα8/rß2 compared to Ldα1/rß2 at same ratios of subunit cRNAs. The sensitivities of neonicotinoids tested for Ldα1/rß2 and Ldα1/Ldα8/rß2 at cRNA ratios of 5:1, 1:1 and 5:5:1, 1:1:1, respectively, were lower than those for nAChRs at ratios of 1:5 and 1:1:5, respectively, whereas the values of maximum response (Imax ) varied. For Ldα1/Ldα8/rß2, a reduction of Lda8 cRNA resulted in increased sensitivity to IMI and decreased sensitivity to TMX. The expression of Ldα1 and Ldα8 significantly decreased in adults by 82.12% and 47.02%, respectively, while Ldα8 was significantly upregulated by 2.44 times in 4th instar larvae after exposure to TMX. We infer that Ldα1 and Ldα8 together play an important role in the sensitivity of L. decemlineata to neonicotinoids.

Rapalogues as hCES2A Inhibitors: In Vitro and In Silico Investigations.

BACKGROUND AND OBJECTIVE: Rapamycin and its semi-synthetic analogues (rapalogues) are frequently used in combination with other prescribed medications in clinical settings. Although the inhibitory effects of rapalogues on cytochrome P450 enzymes (CYPs) have been well examined, the inhibition potentials of rapalogues on human esterases have not been investigated. Herein, the inhibition potentials and inhibitory mechanisms of six marketed rapalogues on human esterases are investigated. METHODS: The inhibitory effects of six marketed rapalogues (rapamycin, zotarolimus, temsirolimus, everolimus, pimecrolimus and tacrolimus) on three major esterases, including human carboxylesterases 1 (hCES1A), human carboxylesterases 2 (hCES2A) and butyrylcholinesterase (BuChE), were assayed using isozyme-specific substrates. Inhibition kinetic analyses and docking simulations were performed to investigate the inhibitory mechanisms of the rapalogues with strong hCES2A inhibition potency. RESULTS: Zotarolimus and pimecrolimus displayed strong inhibition of human hCES2A but these agents did not inhibit hCES1A or BuChE. Further investigation demonstrated that zotarolimus could strongly inhibit intracellular hCES2A in living HepG2 cells, with an estimated IC50 value of 4.09 µM. Inhibition kinetic analyses revealed that zotarolimus inhibited hCES2A-catalyzed fluorescein diacetate hydrolysis in a mixed manner, with the Ki value of 1.61 µM. Docking simulations showed that zotarolimus could tightly bind on hCES2A at two district ligand-binding sites, consistent with its mixed inhibition mode. CONCLUSION: Our findings demonstrate that several marketed rapalogues are potent and specific hCES2A inhibitors, and these agents can serve as leading compounds for the development of more efficacious hCES2A inhibitors to modulate the pharmacokinetic profiles and toxicity of hCES2A-substrate drugs (such as the anticancer agent irinotecan).

Inhibition of human thrombin by the constituents of licorice: inhibition kinetics and mechanistic insights through in vitro and in silico studies.

Thrombin inhibition therapy is a practical strategy to reduce thrombotic and cardiovascular risks via blocking the formation of blood clots. This study aimed to identify naturally occurring thrombin inhibitors from licorice (one of the most popular edible herbs), as well as to investigate their inhibitory mechanisms. Among all tested licorice constituents, licochalcone A was found as the most efficacious agent against human thrombin (IC50 = 7.96 µM). Inhibition kinetic analyses demonstrated that licochalcone A was a mixed inhibitor against thrombin-mediated Z-Gly-Gly-Arg-AMC acetate hydrolysis, with a K i value of 12.23 µM. Furthermore, mass spectrometry-based chemoproteomic assays and molecular docking simulations revealed that licochalcone A could bind to human thrombin at both exosite I and the catalytic site. In summary, our findings demonstrated that the chalcones isolated from licorice were a new class of direct thrombin inhibitors, also suggesting that licochalcone A was a promising lead compound for developing novel anti-thrombotic agents.

The Chromosome-Based Rubber Tree Genome Provides New Insights into Spurge Genome Evolution and Rubber Biosynthesis.

The rubber tree, Hevea brasiliensis, produces natural rubber that serves as an essential industrial raw material. Here, we present a high-quality reference genome for a rubber tree cultivar GT1 using single-molecule real-time sequencing (SMRT) and Hi-C technologies to anchor the ∼1.47-Gb genome assembly into 18 pseudochromosomes. The chromosome-based genome analysis enabled us to establish a model of spurge chromosome evolution, since the common paleopolyploid event occurred before the split of Hevea and Manihot. We show recent and rapid bursts of the three Hevea-specific LTR-retrotransposon families during the last 10 million years, leading to the massive expansion by ∼65.88% (∼970 Mbp) of the whole rubber tree genome since the divergence from Manihot. We identify large-scale expansion of genes associated with whole rubber biosynthesis processes, such as basal metabolic processes, ethylene biosynthesis, and the activation of polysaccharide and glycoprotein lectin, which are important properties for latex production. A map of genomic variation between the cultivated and wild rubber trees was obtained, which contains ∼15.7 million high-quality single-nucleotide polymorphisms. We identified hundreds of candidate domestication genes with drastically lowered genomic diversity in the cultivated but not wild rubber trees despite a relatively short domestication history of rubber tree, some of which are involved in rubber biosynthesis. This genome assembly represents key resources for future rubber tree research and breeding, providing novel targets for improving plant biotic and abiotic tolerance and rubber production.

MLN4924 suppresses neddylation and induces cell cycle arrest, senescence, and apoptosis in human osteosarcoma.

Neddylation is a post-translational protein modification process associated with carcinogenesis and cancer development. MLN4924, a pharmaceutical neddylation inhibitor, induces potent anti-cancer effects in multiple types of cancers. In this study, we investigated the effects of MLN4924 on human osteosarcoma (OS). Levels of both NEDD8 activating enzyme E1 (NAE1) and ubiquitin-conjugating enzyme E2M (Ube2M), two critical components of the neddylation pathway, were much higher in OS tissues and cells than in normal osseous tissues and cells. MLN4924 treatment led to DNA damage, reduced cell viability, senescence and apoptosis in OS cells. Moreover, MLN4924 inhibited OS xenograft tumor growth in mice. Mechanistically, MLN4924 blocked the neddylation of cullins and induced accumulation of several tumor-suppressive substrates of Cullin-RING E3 ubiquitin ligases (CRLs), including CDT1, Wee1, p21, p27, Noxa, and p16. These results suggest clinical studies investigating the utility of MLN4924 for the treatment of OS are warranted.