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Macroautophagy/autophagy plays a crucial role in inhibiting viral replication and regulating the host's immune response. The autophagy receptor SQSTM1/p62 (sequestosome 1) restricts viral replication by directing specific viral proteins to phagophores for degradation. In this study, we investigate the reciprocal relationship between Zika virus (ZIKV) and selective autophagy mediated by SQSTM1/p62. We show that NS2B3 protease encoded by ZIKV cleaves human SQSTM1/p62 at arginine 265 (R265). This cleavage also occurs with endogenous SQSTM1 in ZIKV-infected cells. Furthermore, overexpression of SQSTM1 inhibits ZIKV replication in A549 cells, while its absence increases viral titer. We have also shown that SQSTM1 impedes ZIKV replication by interacting with NS3 and NS5 and directing them to autophagic degradation, and that NS2B3-mediated cleavage could potentially alter this antiviral function of SQSTM1. Taken together, our study highlights the role of SQSTM1-mediated selective autophagy in the host's antiviral defense against ZIKV and uncovers potential viral evasion strategies that exploit the host's autophagic machinery to ensure successful infection. Abbreviation: Cas9: CRISPR-associated protein 9; Co-IP: co-immunoprecipitation; CRISPR: clustered regularly interspaced short palindromic repeats; DENV: dengue virus; GFP: green fluorescent protein; IFA: indirect immunofluorescence assay; KIR: KEAP1-interacting region; KO: knockout; LIR: MAP1LC3/LC3-interacting region; mAb: monoclonal antibody; NBR1: NBR1 autophagy cargo receptor; OPTN: optineurin; pAb: polyclonal antibody; PB1: Phox/BEM1 domain; R265A, a SQSTM1 construct with the arginine (R) residue at position 265 replaced with glutamic acid (A); SQSTM1: sequestosome 1; SQSTM1-C, C-terminal fragment of SQSTM1; SQSTM1-N, N-terminal fragment of SQSTM1; SVV: Seneca Valley virus; TAX1BP1: Tax1 binding protein 1; TBD: TRAF6-binding domain; TCID50: 50% tissue culture infective dose; UBA: ubiquitin-associated domain; Ub: ubiquitin; WT: wild type; ZIKV: Zika virus; ZZ: ZZ-type zinc finger domain. [ABSTRACT FROM AUTHOR]

Dual response polyurethane nanocapsules consisting of a hydrophilic core are synthesized via interfacial polyaddition of the diisocyanate monomers and different diols (VA‐060 and glycols) in inverse miniemulsion process. The presence of the water‐soluble NIR dye in the core and azo‐bonds in the polymer shell allows the selective release of encapsulated material triggered by temperature or NIR light. The capsules are characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The capsule degradation under external stimuli, like temperature and NIR light, is confirmed microscopically by TEM. Macroscopic evidence of the capsule cleavage was achieved by the incorporation of the chemical into the capsule core, and subsequent treatment of the capsules with NIR laser in the presence of the suitable reagent outside the capsules. A color‐forming chemical reaction occurred after the shell opening. The reactions were easily detected by visual observation of a color change and by UV–Vis spectroscopy. [ABSTRACT FROM AUTHOR]

Human-to-human transmission of the highly pathogenic Middle East respiratory syndrome coronavirus (MERS-CoV) is currently inefficient. However, there is concern that the virus might mutate and thereby increase its transmissibility and thus pandemic potential. The pandemic SARS-CoV-2 depends on a highly cleavable furin motif at the S1/S2 site of the viral spike (S) protein for efficient lung cell entry, transmission, and pathogenicity. Here, by employing pseudotyped particles, we investigated whether augmented cleavage at the S1/S2 site also increases MERS-CoV entry into Calu-3 human lung cells. We report that polymorphism T746K at the S1/S2 cleavage site or optimization of the furin motif increases S protein cleavage but not lung cell entry. These findings suggest that, unlike what has been reported for SARS-CoV-2, a highly cleavable S1/S2 site might not augment MERS-CoV infectivity for human lung cells. IMPORTANCE The highly cleavable furin motif in the spike protein is required for robust lung cell entry, transmission, and pathogenicity of SARS-CoV-2. In contrast, it is unknown whether optimization of the furin motif in the spike protein of the pre-pandemic MERS-CoV increases lung cell entry and allows for robust human-human transmission. The present study indicates that this might not be the case. Thus, neither a naturally occurring polymorphism that increased MERS-CoV spike protein cleavage nor art ificial optimization of the cleavage site allowed for increased spike-protein-driven entry into Calu-3 human lung cells. [ABSTRACT FROM AUTHOR]

Rural areas have often been labelled by the literature as 'left-behind' areas or 'places that don't matter', implicitly suggesting that residents of these communities feel neglected by political elites. This article studies the rural-urban divide in external political efficacy, which reflects individuals' beliefs about the responsiveness of political elites, while also examining if compositional and contextual factors can explain such a divide. Drawing on data from the European Social Survey, the results reveal a significant rural-urban gap in external efficacy, which is partly explained by differences in the sociodemographic characteristics of rural and urban dwellers, but not by disparities in their evaluation of the provision of basic public services. Notably, this rural-urban gap in external efficacy is substantively smaller in those countries with higher levels of electoral malapportionment that lead to an overrepresentation of rural areas in national parliaments. [ABSTRACT FROM AUTHOR]

RNA modifications, including N-7-methylguanosine (m7G), are pivotal in governing RNA stability and gene expression regulation. The accurate detection of internal m7G modifications is of paramount significance, given recent associations between altered m7G deposition and elevated expression of the methyltransferase METTL1 in various human cancers. The development of robust m7G detection techniques has posed a significant challenge in the field of epitranscriptomics. In this study, we introduce two methodologies for the global and accurate identification of m7G modifications in human RNA. We introduce borohydride reduction sequencing (Bo-Seq), which provides base resolution mapping of m7G modifications. Bo-Seq achieves exceptional performance through the optimization of RNA depurination and scission, involving the strategic use of high concentrations of NaBH4, neutral pH and the addition of 7-methylguanosine monophosphate (m7GMP) during the reducing reaction. Notably, compared to NaBH4-based methods, Bo-Seq enhances the m7G detection performance, and simplifies the detection process, eliminating the necessity for intricate chemical steps and reducing the protocol duration. In addition, we present an antibody-based approach, which enables the assessment of m7G relative levels across RNA molecules and biological samples, however it should be used with caution due to limitations associated with variations in antibody quality between batches. In summary, our novel approaches address the pressing need for reliable and accessible methods to detect RNA m7G methylation in human cells. These advancements hold the potential to catalyse future investigations in the critical field of epitranscriptomics, shedding light on the complex regulatory roles of m7G in gene expression and its implications in cancer biology.

The literature review discusses and systematizes synthetic approaches to medium-sized cycles and macrocycles based on ring expansion reactions of bi- or polycyclic systems via C-N bond cleavage. Ring expansion reactions of bicyclic ammonium salts proceed via thermal decomposition or the action of strong bases. Bi- or polycyclic systems containing a common amine group can be reduced with strong reducing reagents, e.g. lithium aluminum hydride. Ammonium derivatives are much more prone to nucleophilic attack and quite often are used as starting materials for the synthesis of medium-sized cycles. Bicyclic systems containing a common aminal or amidine group are used for the synthesis of medium-sized rings and macrocycles via cleavage of the endocyclic C-N bond. Various methods of their activation and reduction are discussed in the review.

Previous studies in the horse highlight the potential benefit of prolonged in vitro maturation (IVM) (34 h) compared to short IVM (24 h) with or without prior oocyte holding, but little is known about the optimal IVM duration within this interval. To determine the effect of oocyte holding and duration of IVM ranged between 28 and 34 h on nuclear maturation, cleavage, blastocyst formation, and pregnancy rates, a retrospective study was performed in an equine clinical OPU-ICSI setting. The study included data of 2114 aspirated oocytes from 201 OPU-ICSI sessions. Duration of IVM was divided in three different time windows using quartiles, with 465 oocytes (22.0 %) between 28 and 30 h (first quartile), 1078 oocytes (51.0 %) >30 and 31.7 h (second and third quartiles), and 571 oocytes (27.0 %) >31.7 and 34 h (fourth quartile). Using logistic regression models, the effect of duration of IVM with and without holding was tested on nuclear maturation, cleavage, blastocyst, and pregnancy rates. The three IVM intervals did not show differences in nuclear maturation (respectively 64.5 ± 0.48 %, 65.7 ± 0.47 %, and 67.3 ± 0.47 %), cleavage (respectively 59.7 ± 0.49 %, 58.5 ± 0.49 %, and 64.8 ± 0.48 %), blastocyst (respectively 17.5 ± 0.38 %, 19.0 ± 0.39 %, and 20.8 ± 0.41 %) nor pregnancy rates (respectively 65.4 ± 0.49 %, 70.3 ± 0.46 %, and 74.2 % ± 0.44) (P ≥ 0.38). Oocyte holding prior to IVM did not affect the results either (P ≥ 0.15). In conclusion, oocyte holding and IVM duration between 28 and 34h do not significantly affect outcomes, allowing flexibility in the planning of clinical OPU-ICSI in horses. • Oocyte holding and in vitro maturation duration between 28 and 34 h do not affect equine OPU-ICSI outcomes. • Equine OPU-ICSI outcomes were determined for IVM intervals of 28–30h, 30–31.7h and 31.7–34h, with and without prior oocyte holding. • No differences in nuclear maturation, cleavage blastocyst or pregnancy rates were found between the three IVM time windows and prior oocyte holding did not affect these outcomes either. [ABSTRACT FROM AUTHOR]

Abstract Background Enterovirus A71 (EV-A71), as a neurotropic virus, mainly affects infants and young children under the age of 5. EV-A71 infection causes hand-foot-mouth disease and herpetic angina, and even life-threatening neurological complications. However, the molecular mechanism by which EV-A71 induces nervous system damage remains elusive. The viral protease 3C plays an important role during EV-A71 infection and is also a key intersection of virus-host interactions. Previously, we used yeast two-hybrid to screen out the host protein Double-stranded RNA-binding protein Staufen homolog 2 (Stau2), an important member involved in neuronal mRNA transport, potentially interacts with 3C. Methods We used coimmunoprecipitation (Co-IP) and immunofluorescence assay (IFA) to confirm that EV-A71 3C interacts with Stau2. By constructing the mutant of Stau2, we found the specific site where the 3C protease cleaves Stau2. Detection of VP1 protein using Western blotting characterized EV-A71 viral replication, and overexpression or knockdown of Stau2 exhibited effects on EV-A71 replication. The effect of different cleavage products on EV-A71 replication was demonstrated by constructing Stau2 truncates. Results In this study, we found that EV-A71 3C interacts with Stau2. Stau2 is cleaved by 3C at the Q507-G508 site. Overexpression of Stau2 promotes EV-A71 VP1 protein expression, whereas depletion of Stau2 by small interfering RNA inhibits EV-A71 replication. Stau2 is essential for EV-A71 replication, and the product of Stau2 cleavage by 3C, 508–570 aa, has activity that promotes EV-A71 replication. In addition, we found that mouse Stau2 is also cleaved by EV-A71 3C at the same site. Conclusions Our research provides an example for EV-A71-host interaction, enriching key targets of host factors that contribute to viral replication.

AbstractPlatelet-derived growth factor (PDGF)-induced signalling via PDGF receptor β (PDGFRβ) leads to activation of downstream signalling pathways which regulate multiple cellular responses. It is unclear how PDGFRβ is degraded; both lysosomal and proteasomal degradation have been suggested. In this study, we have characterised the proteolytic cleavage of ligand-activated PDGFRβ, which results in two fragments: a larger fragment containing the extracellular domain, the transmembrane segment, and a part of the intracellular juxtamembrane region with a molecular mass of ∼130 kDa, and an intracellular ∼70 kDa fragment released into the cytoplasm. The proteolytic processing did not take place without internalisation of PDGFRβ. In addition, chelation of intracellular Ca2+ inhibited proteolytic processing. Inhibition of the proteasome affected signal transduction by increasing the phosphorylation of PDGFRβ, PLCγ, and STAT3 while reducing it on Erk1/2 and not affecting Akt. The proteolytic cleavage was observed in fibroblasts or cells that had undergone epithelial-mesenchymal transition. [ABSTRACT FROM AUTHOR]

Dimethylhydrazones of β-dimethylaminomethyl-substituted carbonyl compounds treated with methyl iodide underwent chemoselective methylation at the trialkylamino group. Subsequent treatment of the resulting iodomethylates with bases yielded dimethylhydrazones of α,β-unsaturated enals or enones in high yields. For unsaturated hydrazones, which were stable to bases, the optimal cleaving agent was an aqueous solution of KOH; if labile trimethylsilyl groups were present in the substrate, the best results were obtained with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). [ABSTRACT FROM AUTHOR]

The non-ideal accuracy and insufficient selectivity of CRISPR/Cas9 systems is a serious problem for their use as a genome editing tool. It is important to select the target sequence correctly so that the CRISPR/Cas9 system does not cut similar sequences. This requires an understanding of how and why mismatches in the target sequence can affect the efficiency of the Cas9/sgRNA complex. In this work, we studied the catalytic activity of the Cas9 enzyme to cleave DNA substrates containing nucleotide mismatch at different positions relative to the PAM in the "seed" sequence. We show that mismatches in the complementarity of the sgRNA/DNA duplex at different positions relative to the protospacer adjacent motif (PAM) sequence tend to decrease the cleavage efficiency and increase the half-maximal reaction time. However, for two mismatches at positions 11 and 20 relative to the PAM, an increase in cleavage efficiency was observed, both with and without an increase in half-reaction time. Thermodynamic parameters were obtained from molecular dynamics results, which showed that mismatches at positions 8, 11, and 20 relative to the PAM thermodynamically stabilize the formed complex, and a mismatch at position 2 of the PAM fragment exerts the greatest stabilization compared to the original DNA sequence. The weak correlation of the thermodynamic binding parameters of the components of the Cas9/sgRNA:dsDNA complex with the cleavage data of DNA substrates containing mismatches indicates that the efficiency of Cas9 operation is mainly affected by the conformational changes in Cas9 and the mutual arrangement of sgRNA and substrates. [ABSTRACT FROM AUTHOR]

Histones are essential for DNA packaging and undergo post-translational modifications that significantly influence gene regulation. Among these modifications, histone tail cleavage has recently garnered attention despite being less explored. Cleavage by various proteases impacts processes such as stem cell differentiation, aging, infection, and inflammation, though the mechanisms remain unclear. This review delves into recent insights on histone proteolytic cleavage and its epigenetic significance, highlighting how chromatin, which serves as a dynamic scaffold, responds to signals through histone modification, replacement, and ATP-dependent remodeling. Specifically, histone tail cleavage is linked to critical cellular processes such as granulocyte differentiation, viral infection, aging, yeast sporulation, and cancer development. Although the exact mechanisms connecting histone cleavage to gene expression are still emerging, it is clear that this process represents a novel epigenetic transcriptional mechanism intertwined with chromatin dynamics. This review explores known histone tail cleavage events, the proteolytic enzymes involved, their impact on gene expression, and future research directions in this evolving field. [ABSTRACT FROM AUTHOR]

A tool for precise, target-specific, efficient, and affordable genome editing is a dream for many researchers, from those who conduct basic research to those who use it for applied research. Since 2012, we have tool that almost fulfils such requirements; it is based on clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems. However, even CRISPR/Cas has limitations and obstacles that might surprise its users. In this review, we focus on the most frequently used variant, CRISPR/Cas9 from Streptococcus pyogenes , and highlight key factors affecting its mutagenesis outcomes: (i) factors affecting the CRISPR/Cas9 activity, such as the effect of the target sequence, chromatin state, or Cas9 variant, and how long it remains in place after cleavage; and (ii) factors affecting the follow-up DNA repair mechanisms including mostly the cell type and cell cycle phase, but also, for example, the type of DNA ends produced by Cas9 cleavage (blunt/staggered). Moreover, we note some differences between using CRISPR/Cas9 in plants, yeasts, and animals, as knowledge from individual kingdoms is not fully transferable. Awareness of these factors can increase the likelihood of achieving the expected results of plant genome editing, for which we provide detailed guidelines. [ABSTRACT FROM AUTHOR]

Background: Enterovirus A71 (EV-A71), as a neurotropic virus, mainly affects infants and young children under the age of 5. EV-A71 infection causes hand-foot-mouth disease and herpetic angina, and even life-threatening neurological complications. However, the molecular mechanism by which EV-A71 induces nervous system damage remains elusive. The viral protease 3C plays an important role during EV-A71 infection and is also a key intersection of virus-host interactions. Previously, we used yeast two-hybrid to screen out the host protein Double-stranded RNA-binding protein Staufen homolog 2 (Stau2), an important member involved in neuronal mRNA transport, potentially interacts with 3C. Methods: We used coimmunoprecipitation (Co-IP) and immunofluorescence assay (IFA) to confirm that EV-A71 3C interacts with Stau2. By constructing the mutant of Stau2, we found the specific site where the 3C protease cleaves Stau2. Detection of VP1 protein using Western blotting characterized EV-A71 viral replication, and overexpression or knockdown of Stau2 exhibited effects on EV-A71 replication. The effect of different cleavage products on EV-A71 replication was demonstrated by constructing Stau2 truncates. Results: In this study, we found that EV-A71 3C interacts with Stau2. Stau2 is cleaved by 3C at the Q507-G508 site. Overexpression of Stau2 promotes EV-A71 VP1 protein expression, whereas depletion of Stau2 by small interfering RNA inhibits EV-A71 replication. Stau2 is essential for EV-A71 replication, and the product of Stau2 cleavage by 3C, 508–570 aa, has activity that promotes EV-A71 replication. In addition, we found that mouse Stau2 is also cleaved by EV-A71 3C at the same site. Conclusions: Our research provides an example for EV-A71-host interaction, enriching key targets of host factors that contribute to viral replication. [ABSTRACT FROM AUTHOR]

We report the design of a single RNA sequence capable of adopting one of two ribozyme folds and catalyzing the cleavage and/or ligation of the respective substrates. The RNA is able to change its conformation in response to its environment, hence it is called chameleon ribozyme (CHR). Efficient RNA cleavage of two different substrates as well as RNA ligation by CHR is demonstrated in separate experiments and in a one pot reaction. Our study shows that sequence variants of the hairpin ribozyme intersect with the hammerhead ribozyme and that rather short RNA molecules can have comprehensive conformational flexibility, which is an important feature for the emergence of new functional folds in early evolution. [ABSTRACT FROM AUTHOR]

The present experiment was conducted during January, 2018 to March, 2019 in the laboratory, Dept. of Veterinary Physiology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, Assam (781022), India to investigate the effect of epidermal growth factor on in vitro maturation, fertilization and early embryonic development of cattle oocytes. A total of 318 nos. cattle ovaries were collected from slaughter houses for one year and 1089 nos. culturable oocytes were collected. 381 out of 1089 nos. culturable oocytes were subjected to EGF supplemented serum and serum free basic maturation media. The mean percentage of in-vitro maturation of cattle oocytes based on polar body extrusion was found to be significantly higher in EGF supplemented serum free basic maturation media (70.00±14.49) than the serum basic maturation media without EGF (54.17±7.19) respectively. But the mean percentages of in vitro maturation of cattle oocytes based on the cumulus expansion and polar body extrusion were found to be significantly higher in EGF supplemented serum free basic maturation media (77.59±5.48 and 70.00±14.49) than the value recorded in serum free basic maturation media without EGF (64.20±3.77 and 45.95±8.19). The mean cleavage percentages recorded at 4 cell, 8 cell, 16 cell and morula stages of embryos in EGF supplemented serum free culture media were found to be significantly higher than the value recorded in serum culture media without EGF. From the present experiment, it can be inferred that addition of EGF in serum free basic maturation media had little beneficial effect than serum basic maturation media. [ABSTRACT FROM AUTHOR]

Defects, such as scratches, cleavage fracture, and cleavage pit, occur during surface processing and affect the surface integrity of β-Ga2O3(100) substrates. This study analyzes the effects of polishing materials (polyurethane, lead, and copper) and substrate surface temperatures (25, 10, and 0 ℃) on the polishing results to overcome the effects of such defects on the substrates. Vickers indentation tests were used to detect the effects of the material and surface temperature on the hardness of the disc and substrate, respectively. Machining was verified by conducting ball-disk friction wear and polishing experiments. The experimental results indicate that the material of the polished optical disc significantly affected β-Ga2O3(100) during surface processing. The higher the hardness of the polished optical disc, the greater the material removal rate (MRR) of the substrate surface, and more serious the surface cleavage phenomenon. A lead disc with lower hardness could be used to process β-Ga2O3(100) for realizing a higher MRR, thereby effectively avoiding cleavage fractures and obtaining a surface roughness (Ra) and scratch depth of 3.4 nm and 18 nm, respectively. The hardness and mechanical strength of the substrate could be enhanced at low temperatures, and the indentation depth would become smaller under the action of external forces, effectively inhibiting the formation of scratches and pits on the surface of the substrate during polishing. After polishing the substrate at 0 ℃, the Ra, scratch depth, and total number of cleavage pits were 2.2 nm, 14.3 nm, and 246 nm, respectively. Compared with 25℃, Polishing the substrate at 0 ℃ decreased the Ra, scratch depth, and number of cleavage pits by 35.3, 20.6, and 34.9%, respectively. At 0 ℃, the substrate effectively avoided cleavage fracture and inhibited scratch and pit formation, thereby significantly improving the polishing effect. [ABSTRACT FROM AUTHOR]

Human cancers express altered levels of actin-binding cytoskeletal filamin A (FLNA) protein. FLNA in mammals consists of an actin-binding domain at its N-terminus that is followed by 24 immunoglobulin-like repeat modules interrupted by two hinge regions between repeats 15–16 and 23–24. Cleavage of these hinge regions produces a naturally occurring C-terminal 90 kDa fragment of FLNA (FLNACT) that physically interacts with multiple proteins with diverse functions. This cleavage leads to actin cytoskeleton remodeling, which in turn contributes to cellular signaling, nucleocytoplasmic shuttling of transcriptional factors and nuclear receptors, and regulation of their transcriptional activities that are important for initiation and progression of cancers. Therefore, recent studies have proposed blocking FLNA cleavage as a means of cancer therapy. Here, we update how FLNA cleavage has been targeted by different approaches and their potential implications for future treatment of human cancers. [ABSTRACT FROM AUTHOR]

Interhomolog recombination in meiosis is mediated by the Dmc1 recombinase. The Mei5‐Sae3 complex of Saccharomyces cerevisiae promotes Dmc1 assembly and functions with Dmc1 for homology‐mediated repair of meiotic DNA double‐strand breaks. How Mei5‐Sae3 facilitates Dmc1 assembly remains poorly understood. In this study, we created and characterized several mei5 mutants featuring the amino acid substitutions of basic residues. We found that Arg97 of Mei5, conserved in its ortholog, SFR1 (complex with SWI5), RAD51 mediator, in humans and other organisms, is critical for complex formation with Sae3 for Dmc1 assembly. Moreover, the substitution of either Arg117 or Lys133 with Ala in Mei5 resulted in the production of a C‐terminal truncated Mei5 protein during yeast meiosis. Notably, the shorter Mei5‐R117A protein was observed in meiotic cells but not in mitotic cells when expressed, suggesting a unique regulation of Dmc1‐mediated recombination by posttranslational processing of Mei5‐Sae3. [ABSTRACT FROM AUTHOR]

Although many Fertility Centers have adopted day 5 or 6 embryo transfer policy, yet, 30% of embryo transfers in the US are performed on day 3. This is mainly due to concerns related to longer embryo culture effect and higher rates of embryo transfer cancellation on day 5, with no effect on cumulative pregnancy rate. We conducted a retrospective cohort study comparing individual embryo transfer order rank, best embryo for fresh transfer and intention to freeze, of day-3 and day-5 embryos based on their morphology score. Day-3 embryos of each patient were ranked by embryologists for the order of transfer and intention to freeze, based on morphological score, blinded to actual blastulation outcome. The corresponding blastocysts were similarly ranked for the order of transfer and vitrification intention. Ranking was compared to test the predictive value of day-3 morphological assessment. Sixty patients with 784 day-3 embryos were included. There was only a moderate positive significant correlation between ranks on day-3 and ranks on day-5 [ r = 0.662 95% CI (0.611–0.706, p < 0.001)]. Only 25% of the best embryos for transfer on day 3 (rank = 1) were chosen for fresh transfer on day 5. A total of 441 embryos were intended to be frozen on day 3. Of those, 201 were not transferred nor vitrified on day 5–6 (45%), 3.35 embryos per patient. No significant difference was found between average day-3 rank of embryos ranked 1, 2 (3.12 vs 4.12, p = 0.074) and 3 (3.12 vs 4.08, p = 0.082) on day-5–6. To conclude, this study brings a different perspective to the comparison of day 3 and day 5 by following each embryo's putative and actual designation. Day-3 ranking of embryo morphology did not provide a reliable prediction for blastocyst formation, transfer order and vitrification intention, and may support transfer or cryopreservation of blastocysts over cleavage stage embryos. [ABSTRACT FROM AUTHOR]

The expression of eukaryotic genes relies on the precise 3'-terminal cleavage and polyadenylation of newly synthesized pre-mRNA transcripts. Defects in these processes have been associated with various diseases, including cancer. While cancer-focused sequencing studies have identified numerous driver mutations in protein-coding sequences, noncoding drivers – particularly those affecting the cis-elements required for pre-mRNA cleavage and polyadenylation – have received less attention. Here, we systematically analysed somatic mutations affecting 3'UTR polyadenylation signals in human cancers using the Pan-Cancer Analysis of Whole Genomes (PCAWG) dataset. We found a striking enrichment of cancer-specific somatic mutations that disrupt strong and evolutionarily conserved cleavage and polyadenylation signals within tumour suppressor genes. Further bioinformatics and experimental analyses conducted as a part of our study suggest that these mutations have a profound capacity to downregulate the expression of tumour suppressor genes. Thus, this work uncovers a novel class of noncoding somatic mutations with significant potential to drive cancer progression.

This article discusses the use of sodium hypochlorite pentahydrate (NaOCl·5H2O) as an oxidant for the selective transformation of 1,3-dicarbonyls. The authors conducted experiments to determine the optimal conditions for producing carboxylic acids or dichlorinated compounds. They found that the reaction in acetonitrile resulted in carboxylic acids, while the reaction in acetic acid produced dichlorinated compounds. The authors also explored alternative conditions and found similar results. The study contributes to the development of environmentally friendly transformations of 1,3-dicarbonyl compounds. Additionally, the document provides chemical data and experimental results for various compounds, which can be useful for researchers studying these compounds. [Extracted from the article]

Despite growing interest in both party and citizens’ support for the EU, the relationship between sources of public attitudes toward the EU and the values deriving from structural cleavages remains poorly observed. Particularly, the emergence of a new Transnational Cleavage appears theoretically and empirically disconnected from the state of knowledge on the sources of support and opposition for the Union. How do past explanations of support for the EU relate to individual positioning along Green-Alternative-Libertarian (GAL) and Traditional-Authoritarian-Nationalist (TAN) values? Our research aims to bridge public opinion studies on support for the EU with cleavage theory, going beyond the analysis of party positions, but focusing, instead, on citizens’ beliefs at the individual level. Our results show that transnational cleavage defines a dimension on which explanatory factors of support for the EU align consistently. Differently from the U-curve of the left-right cleavage, it shows a linear trend profiling pros versus opponents of the EU integration process. [ABSTRACT FROM AUTHOR]

A novel fluorogenic alkyl diazirine photocrosslinker bearing an o‐hydroxycinnamate moiety has been developed for identification of the targets of bioactive molecules. The o‐hydroxycinnamate moiety can be converted to the corresponding 7‐hydroxycoumarin derivative, which should be created on the interacting site within the photocaptured target protein. The label yield and fluorescence intensity have been immensely improved in comparison with our previous aromatic crosslinkers to facilitate target identification in small quantities. [ABSTRACT FROM AUTHOR]

This study sought to investigate the synthesis of eight complexes constituted by a cobalt(III) (CoIII) metallic center coordinated to two units of iminic ligands LnC (n = 1–4, L1C–L4C), which are derivatives of pyridoxal hydrochloride and anilines with thioether function containing one to four carbons. Depending on the source of the cobalt ion and the addition (or not) of a non-coordinating counterion, complexes with distinct structures may form, being categorized into two series: [CoIII(LnC)(L0C)] (n = 1–4, C1'–C4') with a LnC ligand and a ligand that has a thiolate function which cleaves the C-S(thioether) bond (L0C) and [CoIII(LnC)2]PF6 (n = 1–4, C1–C4) with two similar units of the same LnC ligand. The occurrence (or not) of cleavage in the eight complexes was observed by elucidating the solid-state structures by single crystal X-ray diffraction. This exciting method allows the synthesis of CoIII complexes without cleaving the C-S bonds from the ligands, thereby not requiring an inert atmosphere in the reaction systems. The synthesized complexes were evaluated by in silico calculations on viable biological targets such as deoxyribonucleic acid, superoxide dismutase enzyme, human serum albumin, and the structural spike glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with the receptor binding domain (RBD) in both up and down conformations without and in complex with the cellular receptor angiotensin-converting enzyme 2 (ACE2). Overall, in silico results suggested that all the inorganic complexes under study are potential anticancer/antiviral agents; however, C4 and C4' are the best candidates for future in vitro assays. [ABSTRACT FROM AUTHOR]

Camellia oleifera is a native woody oil plant in southern China and is infected with anthracnose wherever it is grown. We previously identified Colletotrichum fructicola as the major causal agent of anthracnose on C. oleifera and found that CfAtg8 regulates the pathogenicity and development of C. fructicola. Here, we revealed that CfAtg4 interacts with CfAtg8, contributing to the formation of autophagosomes. The CfAtg81–160 allele, which only contains 1–160 amino acids of the CfAtg8, partially recovered the autophagosome numbers and autophagy defects of the ΔCfatg4 mutant. Consequently, these recoveries resulted in the restoration of the defects of the ΔCfatg4 mutant in growth and responses to different external stresses, albeit to an extent. Importantly, we illustrated the critical roles of CfAtg81–160 in appressoria formation, and pathogenicity. Collectively, our findings provide new insights into the importance of the interaction between CfAtg8 and CfAtg4 in the growth, autophagy and pathogenicity of the phytopathogenic fungi. [ABSTRACT FROM AUTHOR]

The epithelial Na+ channel (ENaC) γ subunit is essential for homeostasis of Na+, K+, and body fluid. Dual γ subunit cleavage before and after a short inhibitory tract allows dissociation of this tract, increasing channel open probability (PO), in vitro. Cleavage proximal to the tract occurs at a furin recognition sequence (143RKRR146, in the mouse γ subunit). Loss of furin-mediated cleavage prevents in vitro activation of the channel by proteolysis at distal sites. We hypothesized that 143RKRR146 mutation to 143QQQQ146 (γQ4) in 129/Sv mice would reduce ENaC PO, impair flow-stimulated flux of Na+ (JNa) and K+ (JK) in perfused collecting ducts, reduce colonic amiloride-sensitive short-circuit current (ISC), and impair Na+, K+, and body fluid homeostasis. Immunoblot of γQ4/Q4 mouse kidney lysates confirmed loss of a band consistent in size with the furin-cleaved proteolytic fragment. However, γQ4/Q4 male mice on a low Na+ diet did not exhibit altered ENaC PO or flow-induced JNa, though flow-induced JK modestly decreased. Colonic amiloride-sensitive ISC in γQ4/Q4 mice was not altered. γQ4/Q4 males, but not females, exhibited mildly impaired fluid volume conservation when challenged with a low Na+ diet. Blood Na+ and K+ were unchanged on a regular, low Na+, or high K+ diet. These findings suggest that biochemical evidence of γ subunit cleavage should not be used in isolation to evaluate ENaC activity. Furthermore, factors independent of γ subunit cleavage modulate channel PO and the influence of ENaC on Na+, K+, and fluid volume homeostasis in 129/Sv mice, in vivo. NEW & NOTEWORTHY: The epithelial Na+ channel (ENaC) is activated in vitro by post-translational proteolysis. In vivo, low Na+ or high K+ diets enhance ENaC proteolysis, and proteolysis is hypothesized to contribute to channel activation in these settings. Using a mouse expressing ENaC with disruption of a key proteolytic cleavage site, this study demonstrates that impaired proteolytic activation of ENaC's γ subunit has little impact upon channel open probability or the ability of mice to adapt to low Na+ or high K+ diets. [ABSTRACT FROM AUTHOR]

Summary: The aim was to explore whether the time-lapse imaging system can help day-3 single cleavage embryo transfer to obtain comparative clinical outcomes to day-4 or 5. The data of 1237 patients who underwent single embryo transfer from January 1, 2018, to September 30, 2020, in our reproductive medicine centre were retrospectively analysed. They were divided into the day-3 single cleavage-stage embryo transfer (SCT) group (n = 357), day-4 single morula transfer (SMT) group (n = 129) and day-5 single blastocyst transfer (SBT) group (n = 751) according to the different embryo transfer stage. The clinical and perinatal outcomes of the three groups were analysed and compared. The clinical pregnancy rates of the patients in the day-3 SCT group, day-4 SMT group and day-5 SBT group were 68.07, 70.54 and 72.04%, respectively. The live birth rates were 56.86, 61.24 and 60.99%, respectively. The monozygotic twin (MZT) rate in the day-3 SCT group was significantly lower than that in the day-5 SBT group (P = 0.049). Regarding perinatal outcomes, only the secondary sex ratio had a significant difference (P < 0.05). After age stratification, no improvement was found in the pregnancy outcomes of patients >35 years of age receiving blastocyst transfer. Our findings suggest that for patients with multiple high-quality embryos on day-3, prolonging the culture time can improve the pregnancy outcome to some extent, but it will bring risks. For centres that have established morphodynamic models, day-3 SCT can also achieve an ideal pregnancy outcome and reduce the rate of monozygotic twins and sex ratio. [ABSTRACT FROM AUTHOR]

SARS-CoV-2 is the pathogen responsible for the most recent global pandemic, which has claimed hundreds of thousands of victims worldwide. Despite remarkable efforts to develop an effective vaccine, concerns have been raised about the actual protection against novel variants. Thus, researchers are eager to identify alternative strategies to fight against this pathogen. Like other opportunistic entities, a key step in the SARS-CoV-2 lifecycle is the maturation of the envelope glycoprotein at the RARR685↓ motif by the cellular enzyme Furin. Inhibition of this cleavage greatly affects viral propagation, thus representing an ideal drug target to contain infection. Importantly, no Furin-escape variants have ever been detected, suggesting that the pathogen cannot replace this protease by any means. Here, we designed a novel fluorogenic SARS-CoV-2-derived substrate to screen commercially available and custom-made libraries of small molecules for the identification of new Furin inhibitors. We found that a peptide substrate mimicking the cleavage site of the envelope glycoprotein of the Omicron variant (QTQTKSHRRAR-AMC) is a superior tool for screening Furin activity when compared to the commercially available Pyr-RTKR-AMC substrate. Using this setting, we identified promising novel compounds able to modulate Furin activity in vitro and suitable for interfering with SARS-CoV-2 maturation. In particular, we showed that 3-((5-((5-bromothiophen-2-yl)methylene)-4-oxo-4,5 dihydrothiazol-2-yl)(3-chloro-4-methylphenyl)amino)propanoic acid (P3, IC50 = 35 μM) may represent an attractive chemical scaffold for the development of more effective antiviral drugs via a mechanism of action that possibly implies the targeting of Furin secondary sites (exosites) rather than its canonical catalytic pocket. Overall, a SARS-CoV-2-derived peptide was investigated as a new substrate for in vitro high-throughput screening (HTS) of Furin inhibitors and allowed the identification of compound P3 as a promising hit with an innovative chemical scaffold. Given the key role of Furin in infection and the lack of any Food and Drug Administration (FDA)-approved Furin inhibitor, P3 represents an interesting antiviral candidate. [ABSTRACT FROM AUTHOR]

The Internet has become central to economic exchange and political communication, placing regulatory initiatives high on European policy agendas. What cleavages shape the political conflicts surrounding Internet policy? I argue that proposals to regulate the Internet frequently affect not only economic interests but also the civil liberties of citizens in the online environment. Political parties must therefore balance their stance on market regulation and their socio-cultural preferences on the 'liberal-authoritarian' dimension of political contestation. To explore party competition on Internet policy in the European Union, I analyse all Internet policy roll-call votes in the European Parliament from 1999 to 2014. Ideal point estimation shows that political competition in this policy field is best explained by the 'liberal-authoritarian' dimension. Reinforcing this finding, two case studies illustrate how civil liberty concerns motivate left-wing parties and the liberal party group to form voting coalitions despite diverging economic preferences. [ABSTRACT FROM AUTHOR]

Objective: To discuss the regulatory effect of serum and glucocorticoid-induced protein kinase 1 (SGK1) in the early development of fertilized eggs at G1 phase of the mice, and to clarify the related mechanism. Methods: Some female mice aged 4-6 weeks and weighed about 20 g, and several male mice aged over 8 weeks and weighed about 30 g were selected. The female mice were intraperitoneally injected with 10 IU of pregnant mare serum gonadotropin (PMSG), followed by 10 IU of human chorionic gonadotropin (HCG) after 48 h. After HCG injection, the female mice were caged overnight with the male mice at a ratio of 1: 1. The fertilized eggs at G1, S, G2, and M phases were collected at 12-21 h, 21-26 h, 26-28 h, and 28-30 h after injected with HCG, and their cellular morphology at different cell cycles were observed under light microscope. The mouse fertilized eggs at G1 phase after superovulation were collected, the mRNA was synthesized in vitro, and divided into no injection group, Tris-EDTA buffer injection group (TE injection group), and SGK1-mRNA injection group. The SGK1 antibodies were mixed with KSOM culture medium with the concentrations of 1 : 25, 1 : 50, 1 : 100, 1: 200, and 0 to culture the mouse fertilized eggs at G1 phase. Western blotting method was used to detect the expression levels of SGK1 protein in fertilized eggs of the mice in various groups and the dephosphorylation for phosphorylated SGK1-Threonine 256 site tyrosine15 site of cell diusion cyclin 2 (Cdc2) (Cdc2-pTyr15) in the fertilized eggs of the mice in various groups and different concentrations of SGK1 antibody groups and the developmental states of the fertilized eggs in the fertilized eggs of the mice in various groups and different concentrations of SGK1 antibody groups were observed under phase contrast microscope; the expression levels of phosphorylated SGK1-Thr256 (SGK1-pThr256) and Cdc2-pTyr15 proteins in fertilized eggs at different post-HCG injection times were detected by Western blotting method. Results: Compared with no injection and TE injection groups, the expression level of SGK1 protein in the cells in SGK1-mRNA injection group was significantly increased (P<0. 01). 27-28 h after injected with HCG, the phosphorylation signaling of Cdc2-pTyr15 in fertilized eggs of the mice in SGK1-mRNA injection group was gradually disappeared, and there was no phosphorylation signaling 29 h after injected with HCG. At 28-29 h after injected with HCG, the phosphorylation signaling of Cdc2-pTyr15 in fertilized eggs of the mice in no injection and TE injection groups gradually disappeared, completely disappeared at 30 h after injected with HCG. With the increasing of the concentration of SGK1 antibody, the disappearing time of the Cdc2-pTyr15 phosphorylation signaling was increased. At 27 h after injected with HCG, the fertilized eggs of the mice in SGK1-mRNA injection group was initiated cleavage; at 31 h after injected with HCG, nearly all the fertilized eggs turned into G2 phase; at 33 h after injected with HCG, all the fertilized eggs in 0 and 1: 200 SGK1 antibody groups underwent cleavage. However, with the increasing of SGK1 antibody concentration, the cleavage of the fertilized eggs in 1: 25, 1: 50, and 1: 100 SGK1 antibody groups was gradually decreased, particularly at 1: 25 SGK1 antibody group. Compared with no injection and TE injection groups, the death rate of the fertilized eggs of the mice in SGK1-mRNA injection group was significantly decreased at 31 h after injected with HCG (P<0. 05), and the cleavage rate was increased (P<0. 05). With the increasing of the SGK1 antibody concentration, the death rates of the fertilized eggs in different concentrations of SGK1 antibody group were increased (P<0. 05), with the extending of cleavage time was increased, and the cleavage rate of the fertilized eggs was decreased in a dose-dependent manner, and the cleavage rate of fertilized eggs in 1: 25 SGK1 antibody group was the lowest. The expression level of SGK1-pThr256 protein in fertilized eggs of the mice was gradually increased from 27 h after injected with HCG (P<0. 05 or P< 0. 01) in a time-dependent manner; at 28 to 29 h after injected with HCG, the expression levels of Cdc2-Tyr15 protein were gradually decreased (P<0. 05) in a time-dependent manner, and had completely disappeared at 30 h after injected with HCG. Conclusion: Both the over-expression and inhibition of SGK1 can affect the time for the fertilized eggs at G1 phase to entry into M phase, suggesting that SGK1 protein may be one of the regulatory factors in the early development of fertilized eggs at G1 phase of the mice, and it may regulate the development of the fertilized eggs at G1 phase through regulation of Cdc2. [ABSTRACT FROM AUTHOR]

ABSTRACT Members of the gasdermin (GSDM) family are critical for inducing programmable pyroptosis by forming pores on the cell membrane. GSDMB, GSDMC, GSDMD, and GSDME are activated by caspases or granzyme, leading to the release of their autoinhibitory domains. The protease SpeB from group A Streptococcus has been shown to cleave and activate GSDMA-mediated pyroptosis. Meanwhile, African Swine Fever Virus infection regulates pyroptosis by cleaving porcine GSDMA (pGSDMA) via active caspase-3 and caspase-4. However, it is not known whether virus-encoded proteases also target GSDMA. Here, we show that residues 1–252 of pGSDMA (pGSDMA1–252) is the pore-forming fragment that induces lytic cell death and pyroptosis. Interestingly, Seneca Valley Virus (SVV) infection induces the cleavage of both pGSDMA and human GSDMA and suppresses GSDMA-mediated cell death. Mechanistically, SVV protease 3C cleaves pGSDMA between Q187 and G188 to generate a shorter fragment, pGSDMA1–186, which fails to induce lytic cell death and lactate dehydrogenase release. Furthermore, pGSDMA1–186 does not localize to the plasma membrane and does not induce cell death, thereby promoting viral replication by suppressing host immune responses. These studies reveal a sophisticated evolutionary adaptation of SVV to bypass GSDMA-mediated pyroptosis, allowing it to overcome host inflammatory defenses.IMPORTANCEGasdermin A (GSDMA) remains a protein shrouded in mystery, particularly regarding its regulation by virus-encoded proteases. Previous studies have identified human GSDMA (hGSDMA) as a sensor and substrate of the SpeB from group A Streptococcus, which initiates pyroptosis. However, it is not clear if viral proteases also cleave GSDMA. In this study, we show that a fragment of porcine GSDMA (pGSDMA) containing the first 252 residues constitutes the pore-forming domain responsible for inducing lytic cell death and pyroptosis. Interestingly, picornavirus Seneca Valley Virus (SVV) protease 3C cleaves both pGSDMA and hGSDMA, generating a shorter fragment that fails to associate with the plasma membrane and does not induce pyroptosis. This cleavage by SVV 3C suppresses GSDMA-mediated lactate dehydrogenase release, bactericidal activity, and lytic cell death. This study reveals how SVV subverts host inflammatory defense by disrupting GSDMA-induced pyroptosis, thereby advancing our understanding of antiviral immunity and opening avenues for treating GSDMA-associated autoimmune diseases.

ABSTRACT Yersinia pestis has recently evolved into a highly lethal flea-borne pathogen through the pseudogenization of extensive genes and the acquisition of exogenous plasmids. Particularly noteworthy are the newly acquired pPCP1 and pMT1 plasmids, which encode the virulence determinants Pla and Yersinia murine toxin (Ymt), crucial for subcutaneous infection and survival within flea vector of Y. pestis, respectively. This study reveals that Pla can cleave Ymt at K299 both in vivo and in vitro. Y. pestis expressing YmtK299A displays enhanced in vitro biofilm formation and increased blood survival, indicating significant roles of Pla-mediated Ymt cleavage in these phenotypes. Intriguingly, although both the ancestral form of Pla and the prevalent Pla-I259T variant in modern Y. pestis strains are capable of cleaving Ymt at K299, the cleavage efficiency of Pla-I259T is only half that of the ancestral variant. In subcutaneous infection, mice infected with Δymt::ymt-K299A show significantly prolonged survival compared to those infected with Δymt::ymt. Similarly, infection with Δpla::pla-I259T also results in extended survival compared to Δpla::pla infection. These data demonstrate that the I259T substitution of Pla mitigates the enhanced virulence of Y. pestis in mice caused by Pla-mediated Ymt cleavage, thereby prolonging the survival period of infected animals and potentially conferring advantages on the transmission of Y. pestis to the next host. These findings deepen our understanding of the intricate interplay between two newly acquired plasmids and shed light on the positive selection of the Pla-I259T mutation, providing new insights into the virulence dynamics and transmission mechanisms of Y. pestis.IMPORTANCEThe emergence of Y. pestis as a highly lethal pathogen is driven by extensive gene pseudogenization and acquisition of exogenous plasmids pPCP1 and pMT1. However, the interplay between these two plasmids during evolution remains largely unexplored. Our study reveals intricate interactions between Ymt and Pla, two crucial virulence determinants encoded on these plasmids. Pla-mediated cleavage of Ymt significantly decreases Y. pestis survival in mouse blood and enhances its virulence in mice. The prevalent Pla-I259T variant in modern strains displays reduced Ymt cleavage, thereby extending the survival of infected animals and potentially increasing strain transmissibility. Our findings shed light on the nuanced evolution of Y. pestis, wherein reduced cleavage efficiency is a positive selection force, shaping the pathogen's natural trajectory.

Seneca Valley virus (SVV) causes vesicular disease in pigs, posing a threat to global pork production. OPTN (optineurin) is a macroautophagy/autophagy receptor that restricts microbial propagation by targeting specific viral or bacterial proteins for degradation. OPTN is degraded and cleaved at glutamine 513 following SVV infection via the activity of viral 3C protease (3C[pro]), resulting in N-terminal and a C-terminal OPTN fragments. Moreover, OPTN interacts with VP1 and targets VP1 for degradation to inhibit viral replication. The N-terminal cleaved OPTN sustained its interaction with VP1, whereas the degradation capacity targeting VP1 decreased. The inhibitory effect of N-terminal OPTN against SVV infection was significantly reduced, C-terminal OPTN failed to inhibit viral replication, and degradation of VP1 was blocked. The knockdown of OPTN resulted in reduced TBK1 activation and phosphorylation of IRF3, whereas overexpression of OPTN led to increased TBK1-IRF3 signaling. Additionally, the N-terminal OPTN diminished the activation of the type I IFN (interferon) pathway. These results show that SVV 3C[pro] targets OPTN because its cleavage impairs its function in selective autophagy and type I IFN production, revealing a novel model in which the virus develops diverse strategies for evading host autophagic machinery and type I IFN response for survival. Abbreviations: Co-IP: co-immunoprecipitation; GFP-green fluorescent protein; hpi: hours post-infection; HRP: horseradish peroxidase; IFN: interferon; IFNB/IFN-β: interferon beta; IRF3: interferon regulatory factor 3; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MOI: multiplicity of infection; OPTN: optineurin; PBS: phosphate-buffered saline; SVV: Seneca Valley virus; SQSTM1: sequestosome 1; TAX1BP1: Tax1 binding protein 1; TBK1: TANK binding kinase 1; TCID50: 50% tissue culture infectious doses; UBAN: ubiquitin binding in TNIP/ABIN (TNFAIP3/A20 and inhibitor of NFKB/NF-kB) and IKBKG/NEMO; UBD: ubiquitin-binding domain; ZnF: zinc finger. [ABSTRACT FROM AUTHOR]

Recent political changes in established democracies have led to a new cleavage, often described as a juxtaposition of 'winners' and 'losers of globalization'. Despite a growing interest in subjective group membership and identity, previous research has not studied whether individuals actually categorize themselves as globalization winners or losers and what effect this has. Based on survey data from Germany, we report evidence of a division between self‐categorized globalization winners and losers that is partially but not completely rooted in social structure and associated with attitudes towards globalization‐related issues and party choices. We thereby confirm many of the assumptions from prior research – such as that (self‐categorized) losers of globalization tend to hold lower levels of education and lean towards the radical right. At the same time, the self‐categorizations are not merely transmission belts of socio‐structural effects but seem to be politically consequential in their own right. We conclude that the categories of globalization winners and losers have the potential to form part of the identity component of the globalization cleavage and are important for understanding how political entrepreneurs appeal to voters on their side of the new divide. [ABSTRACT FROM AUTHOR]

A large and growing body of research draws attention to the rising salience of socio‐cultural and identitarian issues and, potentially, the emergence of a new political cleavage that divides voters on those issues. However, the micro‐foundations of this transformation are less well understood. Here we take a voter‐perspective to evaluate how party competition has been restructured in the eyes of the voter. We leverage measures of citizens' self‐reported probabilities to vote for alternative political parties in the European Election Study voter surveys between 1999 and 2019 in order to map electoral affinity and opposition among party families. We estimate to what extent spatial location on the economic left–right dimension and the GAL‐TAN dimension explain the patterns that emerge, and how this has changed over time. Our results provide evidence of a substantial shift in voter assessment from party competition structured along the economic left–right dimension to competition structured along the GAL‐TAN dimension. We also find great separation of TAN parties from other parties, with the deepest antipathy between the TAN parties and greens. [ABSTRACT FROM AUTHOR]

Background & Aims: Intrahepatic cholangiocarcinoma (iCCA) is a primary liver tumour, characterized by poor prognosis and lack of effective therapy. The cytoskeleton protein Filamin A (FLNA) is involved in cancer progression and metastasis, including primary liver cancer. FLNA is cleaved by calpain, producing a 90 kDa fragment (FLNACT) that can translocate to the nucleus and inhibit gene transcription. We herein aim to define the role of FLNA and its cleavage in iCCA carcinogenesis. Methods & Results: We evaluated the expression and localization of FLNA and FLNACT in liver samples from iCCA patients (n = 82) revealing that FLNA expression was independently correlated with disease-free survival. Primary tumour cells isolated from resected iCCA patients expressed both FLNA and FLNACT, and bulk RNA sequencing revealed a significant enrichment of cell proliferation and cell motility pathways in iCCAs with high FLNA expression. Further, we defined the impact of FLNA and FLNACT on the proliferation and migration of primary iCCA cells (n = 3) and HuCCT1 cell line using silencing and Calpeptin, a calpain inhibitor. We observed that FLNA silencing decreased cell proliferation and migration and Calpeptin was able to reduce FLNACT expression in b oth t he H uCCT1 a nd i CCA cells ( p < .05 vs. control). Moreover, Calpeptin 100 μM decreased HuCCT1 and primary iCCA cell proliferation (p <.00001 vs. control) and migration (p < .05 vs. control). Conclusions: These findings demonstrate that FLNA is involved in human iCCA progression and calpeptin strongly decreased FLNACT expression, reducing cell proliferation and migration. [ABSTRACT FROM AUTHOR]

The effect of cold working on hydrogen embrittlement of Monel-400 alloy has been investigated in this report. Monel-400 alloy having 50 and 60% cold working has been selected for this study. A part of these cold-worked materials is annealed. A series of slow strain rate tensile tests of the cold-worked as well as annealed materials are carried out in two different environmental conditions—air and with hydrogen charging in 3.5% NaCl solution. Alloy with 60% cold working exhibits the highest reduction in ductility with hydrogen charging. The reduction in ductility is lower for the annealed materials compared to that of cold-worked materials when tested with hydrogen charging. The results lead to infer that (i) the alloy becomes more detrimental to hydrogen embrittlement with the increase in percentage cold working, which reduces ductility significantly with increasing strength under hydrogen charging, (ii) annealing of cold-worked materials decreases their susceptibility to hydrogen embrittlement, and (iii) hydrogen damage promotes brittle fracture for the cold-worked materials. [ABSTRACT FROM AUTHOR]

HDAC6, a structurally and functionally unique member of the histone deacetylase (HDAC) family, is an important host factor that restricts viral infection. The broad-spectrum antiviral activity of HDAC6 makes it a potent antiviral agent. Previously, we found that HDAC6 functions to antagonize porcine deltacoronavirus (PDCoV), an emerging enteropathogenic coronavirus with zoonotic potential. However, the final outcome is typically a productive infection that materializes as cells succumb to viral infection, indicating that the virus has evolved sophisticated mechanisms to combat the antiviral effect of HDAC6. Here, we demonstrate that PDCoV nonstructural protein 5 (nsp5) can cleave HDAC6 at glutamine 519 (Q519), and cleavage of HDAC6 was also detected in the context of PDCoV infection. More importantly, the anti-PDCoV activity of HDAC6 was damaged by nsp5 cleavage. Mechanistically, the cleaved HDAC6 fragments (amino acids 1-519 and 520-1159) lost the ability to degrade PDCoV nsp8 due to their impaired deacetylase activity. Furthermore, nsp5-mediated cleavage impaired the ability of HDAC6 to activate RIG-I-mediated interferon responses. We also tested three other swine enteric coronaviruses (transmissible gastroenteritis virus, porcine epidemic diarrhea virus, and swine acute diarrhea syndrome-coronavirus) and found that all these coronaviruses have adopted similar mechanisms to cleave HDAC6 in both an overexpression system and virus-infected cells, suggesting that cleavage of HDAC6 is a common strategy utilized by swine enteric coronaviruses to antagonize the host’s antiviral capacity. Together, these data illustrate how swine enteric coronaviruses antagonize the antiviral function of HDAC6 to maintain their infection, providing new insights to the interaction between virus and host. IMPORTANCE Viral infections and host defenses are in constant opposition. Once viruses combat or evade host restriction, productive infection is achieved. HDAC6 is a broad-spectrum antiviral protein that has been demonstrated to inhibit many viruses, including porcine deltacoronavirus (PDCoV). However, whether HDAC6 is reciprocally targeted and disabled by viruses remains unclear. In this study, we used PDCoV as a model and found that HDAC6 is targeted and cleaved by nsp5, a viral 3C-like protease. The cleaved HDAC6 loses its deacetylase activity as well as its ability to degrade viral proteins and activate interferon responses. Furthermore, this cleavage mechanism is shared among other swine enteric coronaviruses. These findings shed light on the intricate interplay between viruses and HDAC6, highlighting the strategies employed by viruses to evade host antiviral defenses. [ABSTRACT FROM AUTHOR]

Porcine deltacoronavirus (PDCoV) has caused enormous economic losses to the global pig industry. However, the immune escape mechanism of PDCoV remains to be fully clarified. Transcriptomic analysis revealed a high abundance of interferon (IFN)-induced protein with tetratricopeptide repeats 3 (IFIT3) transcripts after PDCoV infection, which initially implied a correlation between IFIT3 and PDCoV. Further studies showed that PDCoV nsp5 could antagonize the host type I interferon signaling pathway by cleaving IFIT3. We demonstrated that PDCoV nsp5 cleaved porcine IFIT3 (pIFIT3) at Gln-406. Similar cleavage of endogenous IFIT3 has also been observed in PDCoV-infected cells. The pIFIT3-Q406A mutant was resistant to nsp5-mediated cleavage and exhibited a greater ability to inhibit PDCoV infection than wild-type pIFIT3. Furthermore, we found that cleavage of IFIT3 is a common characteristic of nsp5 proteins of human coronaviruses, albeit not alphacoronavirus. This finding suggests that the cleavage of IFIT3 is an important mechanism by which PDCoV nsp5 antagonizes IFN signaling. Our study provides new insights into the mechanisms by which PDCoV antagonizes the host innate immune response. IMPORTANCE Porcine deltacoronavirus (PDCoV) is a potential emerging zoonotic pathogen, and studies on the prevalence and pathogenesis of PDCoV are ongoing. The main protease (nsp5) of PDCoV provides an excellent target for antivirals due to its essential and conserved function in the viral replication cycle. Previous studies have revealed that nsp5 of PDCoV antagonizes type I interferon (IFN) production by targeting the interferon-stimulated genes. Here, we provide the first demonstration that nsp5 of PDCoV antagonizes IFN signaling by cleaving IFIT3, which affects the IFN response after PDCoV infection. Our findings reveal that PDCoV nsp5 is an important interferon antagonist and enhance the understanding of immune evasion by deltacoronaviruses [ABSTRACT FROM AUTHOR]

The magnetic-domain structure of the surface of microwires with the composition Fe73.9B13.2Si10.9C2 is studied by magnetic-force microscopy. It is found that removal of the glass shell by chipping leads to distortion of the original magnetic-domain structure. Chemical etching of the glass shell makes it possible to observe the magnetic-domain structure due to the stresses that arise as a result of microwire production. In the absence of an applied magnetic field, a magnetic-domain structure of the surface layer consisting of domain layers inclined to the microwire axis by 45 or 135 degrees is observed. This structure has a close-to-zigzag-like shape. The thickness of the domain layers is not constant and varies from 3 to 5 μm. The application of a constant magnetic field along the microwire axis is found to cause the formation of ring domain layers of various thicknesses (from 1 to 5 μm) with different orientations of the magnetic moment relative to the microwire surface. In a field of 60 Oe along the microwire axis, the magnetic-domain structure consists of only ring layers of domains. Magnetic-field inversion leads to almost complete inversion of the observed domain structure. In this case, complete removal of the magnetic field leads to the formation of a new domain structure of the surface layer. Such a structure is close in shape and position of the domains to the original one, but does not repeat it. [ABSTRACT FROM AUTHOR]