Pannexin-1 channels bridge apoptosis to NLRP3 inflammasome activation.

Apoptosis can promote inflammation by triggering activation of the NLRP3 inflammasome (NLR family, pyrin domain containing 3). However, the molecular mechanisms regulating these processes are ill-defined. We recently reported that pannexin-1 is required to promote NLRP3 inflammasome assembly. We further demonstrate that differential cleavage of gasdermin D (GSDMD) by apoptotic caspases regulates inflammatory cell lysis. Here, we discuss our findings and perspectives for future studies

Arabidopsis Tetraspanins Are Confined to Discrete Expression Domains and Cell Types in Reproductive Tissues and Form Homo- and Heterodimers When Expressed in Yeast

Tetraspanins are evolutionary conserved transmembrane proteins present in all multicellular organisms. In animals, they are known to act as central organizers of membrane complexes and thought to facilitate diverse biological processes, such as cell proliferation, movement, adhesion, and fusion. The genome of Arabidopsis (Arabidopsis thaliana) encodes 17 members of the tetraspanin family; however, little is known about their functions in plant development. Here, we analyzed their phylogeny, protein topology, and domain structure and surveyed their expression and localization patterns in reproductive tissues. We show that, despite their low sequence identity with metazoan tetraspanins, plant tetraspanins display the typical structural topology and most signature features of tetraspanins in other multicellular organisms. Arabidopsis tetraspanins are expressed in diverse tissue domains or cell types in reproductive tissues, and some accumulate at the highest levels in response to pollination in the transmitting tract and stigma, male and female gametophytes and gametes. Arabidopsis tetraspanins are preferentially targeted to the plasma membrane, and they variously associate with specialized membrane domains, in a polarized fashion, to intercellular contacts or plasmodesmata. A membrane-based yeast (Saccharomyces cerevisiae) two-hybrid system established that tetraspanins can physically interact, forming homo- and heterodimer complexes. These results, together with a likely genetic redundancy, suggest that, similar to their metazoan counterparts, plant tetraspanins might be involved in facilitating intercellular communication, whose functions might be determined by the composition of tetraspanin complexes and their binding partners at the cell surface of specific cell types.Marie Curie International Reintegration grant: (no. IRG–256602), U.S. Department of Agriculture-Agricultural Research Service Current Research Information System grant: (5335–21000–030–00D), Fundação Ciência e Tecnologia Postdoctoral Fellowship: (SFRH/BPD/43584/2008), China Scholarship Council fellowship, UC-Berkeley College of Natural Resources SPUR

Physical and dynamical characterization of the Euphrosyne asteroid Family

The Euphrosyne asteroid family occupies a unique zone in orbital element space around 3.15 au and may be an important source of the low-albedo near-Earth objects. The parent body of this family may have been one of the planetesimals that delivered water and organic materials onto the growing terrestrial planets. We aim to characterize the compositional properties as well as the dynamical properties of the family. We performed a systematic study to characterize the physical properties of the Euphrosyne family members via low-resolution spectroscopy using the IRTF telescope. In addition, we performed smoothed-particle hydrodynamics (SPH) simulations and N-body simulations to investigate the collisional origin, determine a realistic velocity field, study the orbital evolution, and constrain the age of the Euphrosyne family. Our spectroscopy survey shows that the family members exhibit a tight taxonomic distribution, suggesting a homogeneous composition of the parent body. Our SPH simulations are consistent with the Euphrosyne family having formed via a reaccumulation process instead of a cratering event. Finally, our N-body simulations indicate that the age of the family is 280 Myr +180/-80 Myr, which is younger than a previous estimate.Comment: 10 pages, 13 figures, accepted to be published in A&

Statistical Mechanics of Nonuniform Magnetization Reversal

The magnetization reversal rate via thermal creation of soliton pairs in quasi-1D ferromagnetic systems is calculated. Such a model describes e.g. the time dependent coercivity of elongated particles as used in magnetic recording media. The energy barrier that has to be overcome by thermal fluctuations corresponds to a soliton-antisoliton pair whose size depends on the external field. In contrast to other models of first order phase transitions such as the phi^4 model, an analytical expression for this energy barrier is found for all values of the external field. The magnetization reversal rate is calculated using a functional Fokker-Planck description of the stochastic magnetization dynamics. Analytical results are obtained in the limits of small fields and fields close to the anisotropy field. In the former case the hard-axis anisotropy becomes effectively strong and the magnetization reversal rate is shown to reduce to the nucleation rate of soliton-antisoliton pairs in the overdamped double sine-Gordon model. The present theory therefore includes the nucleation rate of soliton-antisoliton pairs in the double sine-Gordon chain as a special case. These results demonstrate that for elongated particles, the experimentally observed coercivity is significantly lower than the value predicted by the standard theories of N\'eel and Brown.Comment: 21 pages RevTex 3.0 (twocolumn), 6 figures available on request, to appear in Phys Rev B, Dec (1994

Mobilizable Plasmids for Tunable Gene Expression in Francisella novicida.

Francisella tularensis is the causative agent of the life-threatening disease tularemia. However, the molecular tools to study Francisella are limited. Especially, expression plasmids are sparse and difficult to use, as they are unstable and prone to spontaneous loss. Most Francisella expression plasmids lack inducible promoters making it difficult to control gene expression levels. In addition, available expression plasmids are mainly designed for F. tularensis, however, genetic differences including restriction-modification systems impede the use of these plasmids in F. novicida, which is often used as a model organism to study Francisella pathogenesis. Here we report construction and characterization of two mobilizable plasmids (pFNMB1 and pFNMB2) designed for regulated gene expression in F. novicida. pFNMB plasmids contain a tetracycline inducible promoter to control gene expression levels and oriT for RP4 mediated mobilization. We show that both plasmids are stably maintained in bacteria for more than 40 generations over 4 days of culturing in the absence of selection against plasmid loss. Expression levels are dependent on anhydrotetracycline concentration and homogeneous in a bacterial population. pFNMB1 and pFNMB2 plasmids differ in the sequence between promoter and translation start site and thus allow to reach different maximum levels of protein expression. We used pFNMB1 and pFNMB2 for complementation of Francisella Pathogenicity Island mutants ΔiglF, ΔiglI, and ΔiglC in-vitro and pFNMB1 to complement ΔiglI mutant in bone marrow derived macrophages

Exome sequencing improves the molecular diagnostics of paediatric unexplained neurodevelopmental disorders

\ua9 The Author(s) 2024. Background: Neurodevelopmental disorders (NDDs) and/or associated multiple congenital abnormalities (MCAs) represent a genetically heterogeneous group of conditions with an adverse prognosis for the quality of intellectual and social abilities and common daily functioning. The rapid development of exome sequencing (ES) techniques, together with trio-based analysis, nowadays leads to up to 50% diagnostic yield. Therefore, it is considered as the state-of-the-art approach in these diagnoses. Results: In our study, we present the results of ES in a cohort of 85 families with 90 children with severe NDDs and MCAs. The interconnection of the in-house bioinformatic pipeline and a unique algorithm for variant prioritization resulted in a diagnostic yield of up to 48.9% (44/90), including rare and novel causative variants (41/90) and intragenic copy-number variations (CNVs) (3/90). Of the total number of 47 causative variants, 53.2% (25/47) were novel, highlighting the clinical benefit of ES for unexplained NDDs. Moreover, trio-based ES was verified as a reliable tool for the detection of rare CNVs, ranging from intragenic exon deletions (GRIN2A, ZC4H2 genes) to a 6-Mb duplication. The functional analysis using PANTHER Gene Ontology confirmed the involvement of genes with causative variants in a wide spectrum of developmental processes and molecular pathways, which form essential structural and functional components of the central nervous system. Conclusion: Taken together, we present one of the first ES studies of this scale from the central European region. Based on the high diagnostic yield for paediatric NDDs in this study, 48.9%, we confirm trio-based ES as an effective and reliable first-tier diagnostic test in the genetic evaluation of children with NDDs

Deuteron frozen spin polarized target for nd experiements at the VdG accelerator of Charles University

A frozen spin polarized deuteron target cooled by the 3He/4He dilution refrigerator is described. Fully deuterated 1,2-propanediol was used as a target material. Deuteron vector polarization about 40% was obtained for the target in the shape of a cylinder of 2 cm diameter and 6 cm length. The target is intended for a study of 3N interactions at the polarized neutron beam generated by the Van de Graaff accelerator at the Charles University in Prague

Fluctuations and Instabilities of Ferromagnetic Domain Wall pairs in an External Magnetic Field

Soliton excitations and their stability in anisotropic quasi-1D ferromagnets are analyzed analytically. In the presence of an external magnetic field, the lowest lying topological excitations are shown to be either soliton-soliton or soliton-antisoliton pairs. In ferromagnetic samples of macro- or mesoscopic size, these configurations correspond to twisted or untwisted pairs of Bloch walls. It is shown that the fluctuations around these configurations are governed by the same set of operators. The soliton-antisoliton pair has exactly one unstable mode and thus represents a critical nucleus for thermally activated magnetization reversal in effectively one-dimensional systems. The soliton-soliton pair is stable for small external fields but becomes unstable for large magnetic fields. From the detailed expression of this instability threshold and an analysis of nonlocal demagnetizing effects it is shown that the relative chirality of domain walls can be detected experimentally in thin ferromagnetic films. The static properties of the present model are equivalent to those of a nonlinear sigma-model with anisotropies. In the limit of large hard-axis anisotropy the model reduces to a double sine-Gordon model.Comment: 15 pages RevTex 3.0 (twocolumn), 9 figures available on request, to appear in Phys Rev B, Dec (1994

ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation.

Pyroptosis is a lytic form of cell death that is induced by inflammatory caspases upon activation of the canonical or noncanonical inflammasome pathways. These caspases cleave gasdermin D (GSDMD) to generate an N-terminal GSDMD fragment, which executes pyroptosis by forming membrane pores. We found that calcium influx through GSDMD pores serves as a signal for cells to initiate membrane repair by recruiting the endosomal sorting complexes required for transport (ESCRT) machinery to damaged membrane areas, such as the plasma membrane. Inhibition of the ESCRT-III machinery strongly enhances pyroptosis and interleukin-1β release in both human and murine cells after canonical or noncanonical inflammasome activation. These results not only attribute an anti-inflammatory role to membrane repair by the ESCRT-III system but also provide insight into general cellular survival mechanisms during pyroptosis

AIM2 inflammasome is activated by pharmacological disruption of nuclear envelope integrity.

Inflammasomes are critical sensors that convey cellular stress and pathogen presence to the immune system by activating inflammatory caspases and cytokines such as IL-1β. The nature of endogenous stress signals that activate inflammasomes remains unclear. Here we show that an inhibitor of the HIV aspartyl protease, Nelfinavir, triggers inflammasome formation and elicits an IL-1R-dependent inflammation in mice. We found that Nelfinavir impaired the maturation of lamin A, a structural component of the nuclear envelope, thereby promoting the release of DNA in the cytosol. Moreover, deficiency of the cytosolic DNA-sensor AIM2 impaired Nelfinavir-mediated inflammasome activation. These findings identify a pharmacologic activator of inflammasome and demonstrate the role of AIM2 in detecting endogenous DNA release upon perturbation of nuclear envelope integrity