Influence of different functional elements of plasmid pGT232 on maintenance of recombinant plasmids in Lactobacillus reuteri populations in vitro and in vivo

Plasmid pGT232 (5.1 kb), an indigenous plasmid of Lactobacillus reuteri 100-23, was determined, on the basis of nucleotide and deduced protein sequence data, to belong to the pC194-pUB110 family of plasmids that replicate via the rolling-circle mechanism. The minimal replicon of pGT232 was located on a 1.7-kb sequence consisting of a double-strand origin of replication and a gene encoding the replication initiation protein, repA. An erythromycin-selectable recombinant plasmid containing this minimal replicon was stably maintained (>97% erythromycin-resistant cells) without antibiotic selection in an L. reuteri population under laboratory growth conditions but was poorly maintained (90% resistant cells) of pGT232-derived plasmids in the lactobacillus population in vivo required an additional 1.0-kb sequence which contained a putative single-strand replication origin (SSO). The SSO of pGT232 is believed to be novel and functions in an orientation-specific manner

The β6/β7 region of the Hsp70 substrate-binding domain mediates heat-shock response and prion propagation.

Hsp70 is a highly conserved chaperone that in addition to providing essential cellular functions and aiding in cell survival following exposure to a variety of stresses is also a key modulator of prion propagation. Hsp70 is composed of a nucleotide-binding domain (NBD) and substrate-binding domain (SBD). The key functions of Hsp70 are tightly regulated through an allosteric communication network that coordinates ATPase activity with substrate-binding activity. How Hsp70 conformational changes relate to functional change that results in heat shock and prion-related phenotypes is poorly understood. Here, we utilised the yeast [PSI +] system, coupled with SBD-targeted mutagenesis, to investigate how allosteric changes within key structural regions of the Hsp70 SBD result in functional changes in the protein that translate to phenotypic defects in prion propagation and ability to grow at elevated temperatures. We find that variants mutated within the β6 and β7 region of the SBD are defective in prion propagation and heat-shock phenotypes, due to conformational changes within the SBD. Structural analysis of the mutants identifies a potential NBD:SBD interface and key residues that may play important roles in signal transduction between domains. As a consequence of disrupting the β6/β7 region and the SBD overall, Hsp70 exhibits a variety of functional changes including dysregulation of ATPase activity, reduction in ability to refold proteins and changes to interaction affinity with specific co-chaperones and protein substrates. Our findings relate specific structural changes in Hsp70 to specific changes in functional properties that underpin important phenotypic changes in vivo. A thorough understanding of the molecular mechanisms of Hsp70 regulation and how specific modifications result in phenotypic change is essential for the development of new drugs targeting Hsp70 for therapeutic purposes

Taxonomy and phylogeny of mud owls (Annelida: Sternaspidae), including a new synonymy and new records from the Southern Ocean, North East Atlantic Ocean and Pacific Ocean: challenges in morphological delimitation

Species delimitation in sternaspid polychaetes is currently based on the morphology of a limited suite of characters, namely characters of the ventro-caudal shield—a unique feature of the family. Sternaspid species description has increased rapidly in recent years; however, the validity of the shield as a diagnostic character has not been assessed through molecular means. This study performs the largest molecular taxonomy of Sternaspidae to date, using the nuclear gene 18S, and the mitochondrial genes 16S and cytochrome oxidase subunit I (COI) to assess phylogenetic relationships within the family, to reassess the placement of Sternaspidae within the wider polychaete tree and to investigate the effectiveness of the shield as a diagnostic morphological character. This study includes many new records and reports Sternaspis affinis Stimpson, 1864 from USA Pacific coastline and genetic connectivity between specimens identified as Sternaspis cf. annenkovae Salazar-Vallejo & Buzhinskaja, 2013 from off southeastern Australia and specimens identified as Sternaspis cf. williamsae Salazar-Vallejo & Buzhinskaja, 2013 from the northwestern Pacific. In addition, we investigate material identified as Sternaspis cf. scutata (Ranzani, 1817) in the English Channel and compare with S. scutata through both molecular and morphological means. We further perform a detailed morphological and molecular investigation of new sternaspid material collected from the Southern Ocean and Antarctic Peninsula and regard Sternaspis monroi Salazar-Vallejo, 2014 syn. n. as a junior synonym of Sternaspis sendalli Salazar-Vallejo, 2014, two species recently described from the region, raising questions concerning the validity of current morphological delimitation.© The Author(s) 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made

Exploring the Effect of Crowd Management Measures on Passengers’ Behaviour at Metro Stations

To reduce problems of interaction at the platform train interface (PTI) platform edge doors (PEDs) and markings on the platform are used as door positions indicators. The common methods to study the effect of these measures are based on average values of density using Fruin’s Level of Service (LOS), however identification cannot be made of which part of the PTI is more congested. To solve this problem, a new method is proposed. The method included a conceptual model in which the PTI was discretised into 40 cm square cells to identify which part of the platform is more congested. Passengers’ behaviour was recorded considering two situations before the train arrives: i) passengers waiting in front of the doors; ii) passengers waiting beside the doors. Observation was done at existing stations at Metro de Santiago and London Underground. Results show that PEDs changed the behaviour of passengers as they were located beside the doors rather than in front of them. In addition, when markings were used on the platform, then this behaviour was reinforced. Therefore, it is recommended to use this method to better design the PTI rather than the LOS which is used to design the whole platform. Further research is needed to study the effect of PEDs on passengers with reduced mobility

Does prohibitin expression regulate sperm mitochondrial membrane potential, sperm motility, and male fertility?

Prohibitin (PHB) is a highly conserved major sperm mitochondrial membrane protein whose absence in somatic cells is associated with mitochondrial membrane depolarization and increased generation of reactive oxygen species (ROS). Our recent findings suggest that high levels of oxidants in human semen may contribute to male infertility and that sperm motility could be the earliest and most sensitive indicator of oxidative damage. Based on PHB's roles in mitochondrial sub-compartmentalization and respiratory chain assembly, we examine sperm PHB expression and mitochondrial membrane potential (MITO) in infertile men with poor sperm motility (asthenospermia, A) and/or low sperm concentrations (oligoasthenospermia, OA). Here, we demonstrate that MITO is significantly lower in sperm from A and OA subjects than in normospermic (N) subjects; the decrease is more severe for OA than for A subjects. PHB expression is also significantly lower in sperm from A and OA subjects. Significantly positive correlations are found among PHB expression, MITO, and sperm motility in normospermic, asthenospermic, and oligoasthenospermic subjects. Collectively, our observations lead to the hypothesis that PHB expression is an indicator of sperm quality in infertile men, and that it regulates sperm motility via an alteration in MITO and increased ROS levels. © Copyright 2012, Mary Ann Liebert, Inc.published_or_final_versio

Whole exome sequencing identifies genetic markers of enterovirus susceptibility in East Asians.

INTRODUCTION: Following acute enterovirus (EV) infection, outcomes vary based on factors like the immune response, viral cell entry receptor expression levels, tissue tropism, and genetic factors of both the host and virus. While most individuals exhibit mild, self-limited symptoms, others may suffer severe complications or prolonged infections that can lead to autoimmune disorders. METHODS: To elucidate host responses to EV infection, we performed whole exome sequencing on blood samples from both infected and uninfected individuals. Our initial focus was on genes encoding EV entry receptors-PSGL-1, SCARB2, and ANAXA2 for EV-A71, and CD155 for poliovirus-and on host genes ACBD3 and PI4KΒ, crucial for EV replication. RESULTS: Although no specific genetic variants directly associated with EV infection were identified, we discovered 118 variants across 116 genes enriched in East Asian populations through multi-layered variant filtering. These variants were further analyzed for their potential impacts on organs, biological processes, and molecular pathways. Phenome-wide association studies were conducted to refine our understanding of their contributions to EV infection susceptibility. DISCUSSION: Our findings aim to develop a predictive panel based on these 118 variants, which could help susceptible individuals during EV outbreaks, guiding targeted clinical interventions and preventative strategies

Single valley Dirac fermions in zero-gap HgTe quantum wells

Dirac fermions have been studied intensively in condensed matter physics in recent years. Many theoretical predictions critically depend on the number of valleys where the Dirac fermions are realized. In this work, we report the discovery of a two dimensional system with a single valley Dirac cone. We study the transport properties of HgTe quantum wells grown at the critical thickness separating between the topologically trivial and the quantum spin Hall phases. At high magnetic fields, the quantized Hall plateaus demonstrate the presence of a single valley Dirac point in this system. In addition, we clearly observe the linear dispersion of the zero mode spin levels. Also the conductivity at the Dirac point and its temperature dependence can be understood from single valley Dirac fermion physics.Comment: version 2: supplementary material adde

On the Thermodynamic Geometry and Critical Phenomena of AdS Black Holes

In this paper, we study various aspects of the equilibrium thermodynamic state space geometry of AdS black holes. We first examine the Reissner-Nordstrom-AdS (RN-AdS) and the Kerr-AdS black holes. In this context, the state space scalar curvature of these black holes is analysed in various regions of their thermodynamic parameter space. This provides important new insights into the structure and significance of the scalar curvature. We further investigate critical phenomena, and the behaviour of the scalar curvature near criticality, for KN-AdS black holes in two mixed ensembles, introduced and elucidated in our earlier work arXiv:1002.2538 [hep-th]. The critical exponents are identical to those in the RN-AdS and Kerr-AdS cases in the canonical ensemble. This suggests an universality in the scaling behaviour near critical points of AdS black holes. Our results further highlight qualitative differences in the thermodynamic state space geometry for electric charge and angular momentum fluctuations of these.Comment: 1 + 37 Pages, LaTeX, includes 31 figures. A figure and a clarification added

Strained graphene structures: from valleytronics to pressure sensing

Due to its strong bonds graphene can stretch up to 25% of its original size without breaking. Furthermore, mechanical deformations lead to the generation of pseudo-magnetic fields (PMF) that can exceed 300 T. The generated PMF has opposite direction for electrons originating from different valleys. We show that valley-polarized currents can be generated by local straining of multi-terminal graphene devices. The pseudo-magnetic field created by a Gaussian-like deformation allows electrons from only one valley to transmit and a current of electrons from a single valley is generated at the opposite side of the locally strained region. Furthermore, applying a pressure difference between the two sides of a graphene membrane causes it to bend/bulge resulting in a resistance change. We find that the resistance changes linearly with pressure for bubbles of small radius while the response becomes non-linear for bubbles that stretch almost to the edges of the sample. This is explained as due to the strong interference of propagating electronic modes inside the bubble. Our calculations show that high gauge factors can be obtained in this way which makes graphene a good candidate for pressure sensing.Comment: to appear in proceedings of the NATO Advanced Research Worksho