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| 2. |
- Rohde, M., et al.
(author)
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A blind, numerical benchmark study on supercritical water heat transfer experiments in a 7-rod bundle
- 2016
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In: Journal of Nuclear Engineering and Radiation Science. - : American Society of Mechanical Engineers (ASME). - 2332-8983 .- 2332-8975. ; 2:2
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Journal article (peer-reviewed)abstract
- Heat transfer in supercritical water reactors (SCWRs) shows a complex behavior, especially when the temperatures of the water are near the pseudocritical value. For example, a significant deterioration of heat transfer may occur, resulting in unacceptably high cladding temperatures. The underlying physics and thermodynamics behind this behavior are not well understood yet. To assist the worldwide development in SCWRs, it is therefore of paramount importance to assess the limits and capabilities of currently available models, despite the fact that most of these models were not meant to describe supercritical heat transfer (SCHT). For this reason, the Gen-IV International Forum initiated the present blind, numerical benchmark, primarily aiming to show the predictive ability of currently available models when applied to a real-life application with flow conditions that resemble those of an SCWR. This paper describes the outcomes of ten independent numerical investigations and their comparison with wall temperatures measured at different positions in a 7-rod bundle with spacer grids in a supercritical water test facility at JAEA. The wall temperatures were not known beforehand to guarantee the blindness of the study. A number of models have been used, ranging from a one-dimensional (1-D) analytical approach with heat transfer correlations to a RANS simulation with the SST turbulence model on a mesh consisting of 62 million cells. None of the numerical simulations accurately predicted the wall temperature for the test case in which deterioration of heat transfer occurred. Furthermore, the predictive capabilities of the subchannel analysis were found to be comparable to those of more laborious approaches. It has been concluded that predictions of SCHT in rod bundles with the help of currently available numerical tools and models should be treated with caution.
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| 3. |
- Castanheira, Sonia, et al.
(author)
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A Specialized Peptidoglycan Synthase Promotes Salmonella Cell Division inside Host Cells
- 2017
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In: mBio. - 2161-2129 .- 2150-7511. ; 8:6
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Journal article (peer-reviewed)abstract
- Bacterial cell division has been studied extensively under laboratory conditions. Despite being a key event in the bacterial cell cycle, cell division has not been explored in vivo in bacterial pathogens interacting with their hosts. We discovered in Salmonella enterica serovar Typhimurium a gene absent in nonpathogenic bacteria and encoding a peptidoglycan synthase with 63% identity to penicillin-binding protein 3 (PBP3). PBP3 is an essential cell division-specific peptidoglycan synthase that builds the septum required to separate daughter cells. Since S. Typhimurium carries genes that encode a PBP3 paralog-which we named PBP3(SAL)-and PBP3, we hypothesized that there are different cell division events in host and nonhost environments. To test this, we generated S. Typhimurium isogenic mutants lacking PBP3(SAL) or the hitherto considered essential PBP3. While PBP3 alone promotes cell division under all conditions tested, the mutant producing only PBP3(SAL) proliferates under acidic conditions (pH <= 5.8) but does not divide at neutral pH. PBP3(SAL) production is tightly regulated with increased levels as bacteria grow in media acidified up to pH 4.0 and in intracellular bacteria infecting eukaryotic cells. PBP3(SAL) activity is also strictly dependent on acidic pH, as shown by beta-lactam antibiotic binding assays. Live-cell imaging microscopy revealed that PBP3(SAL) alone is sufficient for S. Typhimurium to divide within phagosomes of the eukaryotic cell. Additionally, we detected much larger amounts of PBP3(SAL) than those of PBP3 in vivo in bacteria colonizing mouse target organs. Therefore, PBP3(SAL) evolved in S. Typhimurium as a specialized peptidoglycan synthase promoting cell division in the acidic intraphagosomal environment. IMPORTANCE During bacterial cell division, daughter cells separate by a transversal structure known as the division septum. The septum is a continuum of the cell wall and therefore is composed of membrane(s) and a peptidoglycan layer. To date, actively growing bacteria were reported to have only a "cell division-specific" peptidoglycan synthase required for the last steps of septum formation and consequently, essential for bacterial life. Here, we discovered that Salmonella enterica has two peptidoglycan synthases capable of synthesizing the division septum. One of these enzymes, PBP3(SAL), is present only in bacterial pathogens and evolved in Salmonella to function exclusively in acidic environments. PBP3(SAL) is used preferentially by Salmonella to promote cell division in vivo in mouse target organs and inside acidified phagosomes. Our data challenge the concept of only one essential cell division-specific peptidoglycan synthase and demonstrate that pathogens can divide in defined host locations using alternative mechanisms.
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| 4. |
- Hernández, Sara B., et al.
(author)
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Bile-induced peptidoglycan remodeling in Salmonella enterica
- 2015
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In: Environmental Microbiology. - : Wiley-Blackwell. - 1462-2912 .- 1462-2920. ; 17:4, s. 1081-1089
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Journal article (peer-reviewed)abstract
- Changes in the peptidoglycan (PG) structure of Salmonella enterica are detected in the presence of a sublethal concentration of sodium deoxycholate (DOC): (i) lower proportions of Braun lipoprotein (Lpp)-bound muropeptides; (ii) reduced levels of muropeptides cross-linked by L(meso)-diaminopimelyl-D(meso)-diaminopimelic acid (L-D) peptide bridges (3-3 cross-links). Similar structural changes are found in S. enterica cultures adapted to grow in the presence of a lethal concentration of DOC, suggesting that reduced anchoring of Braun protein to PG and low occurrence of 3-3 cross-links may increase S. enterica resistance to bile. This view is further supported by additional observations: (i) A triple mutant lacking L,D-transpeptidases YbiS, ErfK, and YcfS, which does not contain Lpp anchored to PG, is hyper-resistant to bile; (ii) enhanced 3-3 cross-linking upon overexpression of YnhG transpeptidase causes a decrease in bile resistance. These observations suggest that remodelling of the cell wall may be added to the list of adaptive responses that permit survival of S. enterica in the presence of bile.
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| 5. |
- Hernández, Sara B., et al.
(author)
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Peptidoglycan editing in non-proliferating intracellular Salmonella as source of interference with immune signaling
- 2022
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In: PLoS Pathogens. - : Public Library of Science. - 1553-7366 .- 1553-7374. ; 18:1
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Journal article (peer-reviewed)abstract
- Salmonella enterica causes intracellular infections that can be limited to the intestine or spread to deeper tissues. In most cases, intracellular bacteria show moderate growth. How these bacteria face host defenses that recognize peptidoglycan, is poorly understood. Here, we report a high-resolution structural analysis of the minute amounts of peptidoglycan purified from S. enterica serovar Typhimurium (S. Typhimurium) infecting fibroblasts, a cell type in which this pathogen undergoes moderate growth and persists for days intracellularly. The peptidoglycan of these non-proliferating bacteria contains atypical crosslinked muropeptides with stem peptides trimmed at the L-alanine-D-glutamic acid-(γ) or D-glutamic acid-(γ)-meso-diaminopimelic acid motifs, both sensed by intracellular immune receptors. This peptidoglycan has a reduced glycan chain average length and ~30% increase in the L,D-cross-link, a type of bridge shared by all the atypical crosslinked muropeptides identified. The L,Dtranspeptidases LdtD (YcbB) and LdtE (YnhG) are responsible for the formation of these L, D-bridges in the peptidoglycan of intracellular bacteria. We also identified in a fraction of muropeptides an unprecedented modification in the peptidoglycan of intracellular S. Typhimurium consisting of the amino alcohol alaninol replacing the terminal (fourth) D-alanine. Alaninol was still detectable in the peptidoglycan of a double mutant lacking LdtD and LdtE, thereby ruling out the contribution of these enzymes to this chemical modification. Remarkably, all multiple mutants tested lacking candidate enzymes that either trim stem peptides or form the L,D-bridges retain the capacity to modify the terminal D-alanine to alaninol and all attenuate NF-κB nuclear translocation. These data inferred a potential role of alaninol-containing muropeptides in attenuating pro-inflammatory signaling, which was confirmed with a synthetic tetrapeptide bearing such amino alcohol. We suggest that the modification of D-alanine to alaninol in the peptidoglycan of non-proliferating intracellular S. Typhimurium is an editing process exploited by this pathogen to evade immune recognition inside host cells.
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