The brain-specific double-stranded RNA-binding protein Staufen2 is required for dendritic spine morphogenesis

Mammalian Staufen2 (Stau2) is a member of the double-stranded RNA-binding protein family. Its expression is largely restricted to the brain. It is thought to play a role in the delivery of RNA to dendrites of polarized neurons. To investigate the function of Stau2 in mature neurons, we interfered with Stau2 expression by RNA interference (RNAi). Mature neurons lacking Stau2 displayed a significant reduction in the number of dendritic spines and an increase in filopodia-like structures. The number of PSD95-positive synapses and miniature excitatory postsynaptic currents were markedly reduced in Stau2 down-regulated neurons. Akin effects were caused by overexpression of dominant-negative Stau2. The observed phenotype could be rescued by overexpression of two RNAi cleavage-resistant Stau2 isoforms. In situ hybridization revealed reduced expression levels of β-actin mRNA and fewer dendritic β-actin mRNPs in Stau2 down-regulated neurons. Thus, our data suggest an important role for Stau2 in the formation and maintenance of dendritic spines of hippocampal neurons

Pulmonary siRNA Delivery with Sophisticated Amphiphilic Poly(Spermine Acrylamides) for the Treatment of Lung Fibrosis

RNA interference (RNAi) is an efficient strategy to post-transcriptionally silence gene expression. While all siRNA drugs on the market target the liver, the lung offers a variety of currently undruggable targets, which can potentially be treated with RNA therapeutics. To achieve this goal, the synthesis of poly(spermine acrylamides) (P(SpAA) is reported herein. Polymers are prepared via polymerization of N-acryloxysuccinimide (NAS) and afterward this active ester is converted into spermine-based pendant groups. Copolymerizations with decylacrylamide are employed to increase the hydrophobicity of the polymers. After deprotection, polymers show excellent siRNA encapsulation to obtain perfectly sized polyplexes at very low polymer/RNA ratios. In vitro 2D and 3D cell culture, ex vivo and in vivo experiments reveal superior properties of amphiphilic spermine-copolymers with respect to delivery of siRNA to lung cells in comparison to commonly used lipid-based transfection agents. In line with the in vitro results, siRNA delivery to human lung explants confirm more efficient gene silencing of protease-activated receptor 2 (PAR2), a G protein-coupled receptor involved in fibrosis. This study reveals the importance of the balance between efficient polyplex formation, cellular uptake, gene knockdown, and toxicity for efficient siRNA delivery in vitro, in vivo, and in fibrotic human lung tissue ex vivo

The serotonin receptor 3E variant is a risk factor for female IBS-D

Irritable bowel syndrome (IBS) is a gut-brain disorder of multifactorial origin. Evidence of disturbed serotonergic function in IBS accumulated for the 5-HT3 receptor family. 5-HT3Rs are encoded by HTR3 genes and control GI function, and peristalsis and secretion, in particular. Moreover, 5-HT3R antagonists are beneficial in the treatment of diarrhea predominant IBS (IBS-D). We previously reported on functionally relevant SNPs in HTR3A c.-42C > T (rs1062613), HTR3C p.N163K (rs6766410), and HTR3E c.*76G > A (rs56109847 = rs62625044) being associated with IBS-D, and the HTR3B variant p.Y129S (rs1176744) was also described within the context of IBS. We performed a multi-center study to validate previous results and provide further evidence for the relevance of HTR3 genes in IBS pathogenesis. Therefore, genotype data of 2682 IBS patients and 9650 controls from 14 cohorts (Chile, Germany (2), Greece, Ireland, Spain, Sweden (2), the UK (3), and the USA (3)) were taken into account. Subsequent meta-analysis confirmed HTR3E c.*76G > A (rs56109847 = rs62625044) to be associated with female IBS-D (OR = 1.58; 95% CI (1.18, 2.12)). Complementary expression studies of four GI regions (jejunum, ileum, colon, sigmoid colon) of 66 IBS patients and 42 controls revealed only HTR3E to be robustly expressed. On top, HTR3E transcript levels were significantly reduced in the sigma of IBS patients (p = 0.0187); more specifically, in those diagnosed with IBS-D (p = 0.0145). In conclusion, meta-analysis confirmed rs56109847 = rs62625044 as a risk factor for female IBS-D. Expression analysis revealed reduced HTR3E levels in the sigmoid colon of IBS-D patients, which underlines the relevance of HTR3E in the pathogenesis of IBS-D

The serotonin receptor 3E variant is a risk factor for female IBS-D

Irritable bowel syndrome (IBS) is a gut-brain disorder of multifactorial origin. Evidence of disturbed serotonergic function in IBS accumulated for the 5-HT receptor family. 5-HT Rs are encoded by HTR3 genes and control GI function, and peristalsis and secretion, in particular. Moreover, 5-HT R antagonists are beneficial in the treatment of diarrhea predominant IBS (IBS-D). We previously reported on functionally relevant SNPs in HTR3A c.-42C > T (rs1062613), HTR3C p.N163K (rs6766410), and HTR3E c.*76G > A (rs56109847 = rs62625044) being associated with IBS-D, and the HTR3B variant p.Y129S (rs1176744) was also described within the context of IBS. We performed a multi-center study to validate previous results and provide further evidence for the relevance of HTR3 genes in IBS pathogenesis. Therefore, genotype data of 2682 IBS patients and 9650 controls from 14 cohorts (Chile, Germany (2), Greece, Ireland, Spain, Sweden (2), the UK (3), and the USA (3)) were taken into account. Subsequent meta-analysis confirmed HTR3E c.*76G > A (rs56109847 = rs62625044) to be associated with female IBS-D (OR = 1.58; 95% CI (1.18, 2.12)). Complementary expression studies of four GI regions (jejunum, ileum, colon, sigmoid colon) of 66 IBS patients and 42 controls revealed only HTR3E to be robustly expressed. On top, HTR3E transcript levels were significantly reduced in the sigma of IBS patients (p = 0.0187); more specifically, in those diagnosed with IBS-D (p = 0.0145). In conclusion, meta-analysis confirmed rs56109847 = rs62625044 as a risk factor for female IBS-D. Expression analysis revealed reduced HTR3E levels in the sigmoid colon of IBS-D patients, which underlines the relevance of HTR3E in the pathogenesis of IBS-D. [Abstract copyright: © 2022. The Author(s).

Genome-wide analysis of 53,400 people with irritable bowel syndrome highlights shared genetic pathways with mood and anxiety disorders

Funder: Kennedy Trust Rheumatology Research Prize StudentshipFunder: DFG Cluster of Excellence “Precision Medicine in Chronic In-flammation” (PMI; ID: EXC2167)Funder: EC | EC Seventh Framework Programm | FP7 Ideas: European Research Council (FP7-IDEAS-ERC - Specific Programme: “Ideas” Implementing the Seventh Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (2007 to 2013)); doi: https://doi.org/10.13039/100011199; Grant(s): 715772Funder: NWO-VIDI grant 016.178.056, the Netherlands Heart Foundation CVON grant 2018-27, and NWO Gravitation grant ExposomeNLFunder: Li Ka Shing Foundation (Li Ka Shing Foundation Limited); doi: https://doi.org/10.13039/100007421Abstract: Irritable bowel syndrome (IBS) results from disordered brain–gut interactions. Identifying susceptibility genes could highlight the underlying pathophysiological mechanisms. We designed a digestive health questionnaire for UK Biobank and combined identified cases with IBS with independent cohorts. We conducted a genome-wide association study with 53,400 cases and 433,201 controls and replicated significant associations in a 23andMe panel (205,252 cases and 1,384,055 controls). Our study identified and confirmed six genetic susceptibility loci for IBS. Implicated genes included NCAM1, CADM2, PHF2/FAM120A, DOCK9, CKAP2/TPTE2P3 and BAG6. The first four are associated with mood and anxiety disorders, expressed in the nervous system, or both. Mirroring this, we also found strong genome-wide correlation between the risk of IBS and anxiety, neuroticism and depression (rg > 0.5). Additional analyses suggested this arises due to shared pathogenic pathways rather than, for example, anxiety causing abdominal symptoms. Implicated mechanisms require further exploration to help understand the altered brain–gut interactions underlying IBS

C–H Bond Activation of Silyl-Substituted Pyridines with Bis(Phenolate)Yttrium Catalysts as a Facile Tool towards Hydroxyl-Terminated Michael-Type Polymers

Herein, silicon-protected, ortho-methylated hydroxy-pyridines were reported as initiators in 2-aminoalkoxy-bis(phenolate)yttrium complexes for rare earth metal-mediated group-transfer polymerization (REM-GTP) of Michael-type monomers. To introduce these initiators, C−H bond activation was performed by reacting [(ONOO)tBuY(X)(thf)] (X = CH2TMS, thf = tetrahydrofuran) with tert-butyl-dimethyl-silyl-functionalized α-methylpyridine to obtain the complex [(ONOOtBuY(X)(thf)] (X = 4-(4′-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)-2,6-di-methylpyridine). These initiators served as functional end-groups in polymers produced via REM-GTP. In this contribution, homopolymers of 2-vinylpyridine (2VP) and diethyl vinyl phosphonate (DEVP) were produced. Activity studies and end-group analysis via mass spectrometry, size-exclusion chromatography (SEC) and NMR spectroscopy were performed to reveal the initiator efficiency, the catalyst activity towards both monomers as well as the initiation mechanism of this initiator in contrast to commonly used alkyl initiators. In addition, 2D NMR studies were used to further confirm the end-group integrity of the polymers. For all polymers, different deprotection routes were evaluated to obtain hydroxyl-terminated poly(2-vinylpyridine) (P2VP) and poly(diethyl vinyl phosphonate) (PDEVP). Such hydroxyl groups bear the potential to act as anchoring points for small bioactive molecules, for post-polymerization functionalization or as macroinitiators for further polymerizations

Detection of the Lateral Thermal Spread during Bipolar Vessel Sealing in an Ex Vivo Model—Preliminary Results

Background: As an unwanted side effect, lateral thermal expansion in bipolar tissue sealing may lead to collateral tissue damage. Materials and Methods: Our investigations were carried out on an ex vivo model of porcine carotid arteries. Lateral thermal expansion was measured and a calculated index, based on thermographic recording and histologic examination, was designed to describe the risk of tissue damage. Results: For instrument 1, the mean extent of the critical zone > 50 °C was 2315 ± 509.2 µm above and 1700 ± 331.3 µm below the branches. The width of the necrosis zone was 412.5 ± 79.0 µm above and 426.7 ± 100.7µm below the branches. For instrument 2, the mean extent of the zone > 50 °C was 2032 ± 592.4 µm above and 1182 ± 386.9 µm below the branches. The width of the necrosis zone was 642.6 ± 158.2 µm above and 645.3 ± 111.9 µm below the branches. Our risk index indicated a low risk of damage for instrument 1 and a moderate to high risk for instrument 2. Conclusion: Thermography is a suitable method to estimate lateral heat propagation, and a validated risk index may lead to improved surgical handling

Azide‐Modified Poly(diethyl vinylphosphonate) for Straightforward Graft‐to Carbon Nanotube Functionalization

Abstract Rare‐earth metal‐mediated group‐transfer polymerization (REM‐GTP) offers distinctive features over common polymerization techniques, such as living character, a broad scope of functional monomers, high activity, excellent control of the polymeric parameters as well as inherent chain‐end functionalization. Through the latter, polymers with reactive end‐groups become feasible, opening the pathway for further post‐polymerization functionalization. In this study, a straightforward graft‐to immobilization of the Michael‐type polymer poly(diethyl vinylphosphonate) (PDEVP) on multi‐walled carbon nanotubes (MWCNT) is reported. Hence, a customized azide initiator is synthesized and studied in the CH bond activation with various lanthanide‐based catalysts and the subsequent polymerization of diethyl vinylphosphonate (DEVP). The successful attachment of the azide end‐group is demonstrated via electrospray ionization mass spectrometry (ESI‐MS) and the synthesized polymers are subjected to immobilization on multi‐walled carbon nanotubes in a graft‐to approach. The prepared MWCNT:PDEVP composites are analyzed via thermogravimetric analysis (TGA), elemental analysis (EA), Raman spectroscopy, X‐Ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM) and the versatility of this approach is shown via the stabilization of MWCNT dispersions in water