43 research outputs found
Recombinant Probiotic Expressing Listeria Adhesion Protein Attenuates Listeria monocytogenes Virulence In Vitro
BACKGROUND: Listeria monocytogenes, an intracellular foodborne pathogen, infects immunocompromised hosts. The primary route of transmission is through contaminated food. In the gastrointestinal tract, it traverses the epithelial barrier through intracellular or paracellular routes. Strategies to prevent L. monocytogenes entry can potentially minimize infection in high-risk populations. Listeria adhesion protein (LAP) aids L. monocytogenes in crossing epithelial barriers via the paracellular route. The use of recombinant probiotic bacteria expressing LAP would aid targeted clearance of Listeria from the gut and protect high-risk populations from infection. METHODOLOGY/PRINCIPAL FINDINGS: The objective was to investigate the ability of probiotic bacteria or LAP-expressing recombinant probiotic Lactobacillus paracasei (Lbp(LAP)) to prevent L. monocytogenes adhesion, invasion, and transwell-based transepithelial translocation in a Caco-2 cell culture model. Several wild type probiotic bacteria showed strong adhesion to Caco-2 cells but none effectively prevented L. monocytogenes infection. Pre-exposure to Lbp(LAP) for 1, 4, 15, or 24 h significantly (P<0.05) reduced adhesion, invasion, and transepithelial translocation of L. monocytogenes in Caco-2 cells, whereas pre-exposure to parental Lb. paracasei had no significant effect. Similarly, Lbp(LAP) pre-exposure reduced L. monocytogenes translocation by as much as 46% after 24 h. Lbp(LAP) also prevented L. monocytogenes-mediated cell damage and compromise of tight junction integrity. Furthermore, Lbp(LAP) cells reduced L. monocytogenes-mediated cell cytotoxicity by 99.8% after 1 h and 79% after 24 h. CONCLUSIONS/SIGNIFICANCE: Wild type probiotic bacteria were unable to prevent L. monocytogenes infection in vitro. In contrast, Lbp(LAP) blocked adhesion, invasion, and translocation of L. monocytogenes by interacting with host cell receptor Hsp60, thereby protecting cells from infection. These data show promise for the use of recombinant probiotics in preventing L. monocytogenes infection in high-risk populations
N-Terminal Gly224–Gly411 Domain in Listeria Adhesion Protein Interacts with Host Receptor Hsp60
Listeria adhesion protein (LAP) is a housekeeping bifunctional enzyme consisting of N-terminal acetaldehyde dehydrogenase (ALDH) and C-terminal alcohol dehydrogenase (ADH). It aids Listeria monocytogenes in crossing the epithelial barrier through a paracellular route by interacting with its host receptor, heat shock protein 60 (Hsp60). To gain insight into the binding interaction between LAP and Hsp60, LAP subdomain(s) participating in the Hsp60 interaction were investigated.Using a ModBase structural model, LAP was divided into 4 putative subdomains: the ALDH region contains N1 (Met(1)-Pro(223)) and N2 (Gly(224)-Gly(411)), and the ADH region contains C1 (Gly(412)-Val(648)) and C2 (Pro(649)-Val(866)). Each subdomain was cloned and overexpressed in Escherichia coli and purified. Purified subdomains were used in ligand overlay, immunofluorescence, and bead-based epithelial cell adhesion assays to analyze each domain's affinity toward Hsp60 protein or human ileocecal epithelial HCT-8 cells.The N2 subdomain exhibited the greatest affinity for Hsp60 with a K(D) of 9.50±2.6 nM. The K(D) of full-length LAP (7.2±0.5 nM) to Hsp60 was comparable to the N2 value. Microspheres (1 µm diameter) coated with N2 subdomain showed significantly (P<0.05) higher binding to HCT-8 cells than beads coated with other subdomains and this binding was inhibited when HCT-8 cells were pretreated with anti-Hsp60 antibody to specifically block epithelial Hsp60. Furthermore, HCT-8 cells pretreated with purified N2 subdomain also reduced L. monocytogenes adhesion by about 4 log confirming its involvement in interaction with epithelial cells.These data indicate that the N2 subdomain in the LAP ALDH domain is critical in initiating interaction with mammalian cell receptor Hsp60 providing insight into the molecular mechanism of pathogenesis for the development of potential anti-listerial control strategies
Supernova remnants: the X-ray perspective
Supernova remnants are beautiful astronomical objects that are also of high
scientific interest, because they provide insights into supernova explosion
mechanisms, and because they are the likely sources of Galactic cosmic rays.
X-ray observations are an important means to study these objects.And in
particular the advances made in X-ray imaging spectroscopy over the last two
decades has greatly increased our knowledge about supernova remnants. It has
made it possible to map the products of fresh nucleosynthesis, and resulted in
the identification of regions near shock fronts that emit X-ray synchrotron
radiation.
In this text all the relevant aspects of X-ray emission from supernova
remnants are reviewed and put into the context of supernova explosion
properties and the physics and evolution of supernova remnants. The first half
of this review has a more tutorial style and discusses the basics of supernova
remnant physics and thermal and non-thermal X-ray emission. The second half
offers a review of the recent advances.The topics addressed there are core
collapse and thermonuclear supernova remnants, SN 1987A, mature supernova
remnants, mixed-morphology remnants, including a discussion of the recent
finding of overionization in some of them, and finally X-ray synchrotron
radiation and its consequences for particle acceleration and magnetic fields.Comment: Published in Astronomy and Astrophysics Reviews. This version has 2
column-layout. 78 pages, 42 figures. This replaced version has some minor
language edits and several references have been correcte
Community-Based Funding and Budgeting: Participatory Budgeting as a Transformative Act
Jung Min Choi, John W. Murphy, Ramsey Dahab, and Charlene Holkenbrink-Monk, in this chapter, explore the issue of funding and budgeting. Often funds are directed to community organizations in ways that are either irrelevant or difficult to use. Additionally, budgets are formulated by agencies that are disconnected from the communities where services are needed. Community-based funding and budgeting, accordingly, are beginning to receive serious attention. Communities, accordingly, are given the latitude in some cases to establish budgets and spending strategies, along with identifying and pursuing sources of funds that are consistent ethically with these priorities and desires. Community-based funding and budgeting, in this way, are vital to supporting interventions in a community-sensitive manner