scholarly journals The Legionella Lqs-LvbR regulatory network controls temperature-dependent growth onset and bacterial cell density

2021 ◽  
Author(s):  
Ramon Hochstrasser ◽  
Hubert Hilbi
Keyword(s):  
Cell Density ◽  
Legionella Pneumophila ◽  
Elevated Temperatures ◽  
Optimal Growth ◽  
Lag Phase ◽  
Temperature Dependent ◽  
Genes Encoding ◽  
Life Threatening ◽  
Cell Densities ◽  
Dependent Growth

Legionella species are facultative intracellular pathogens, which can cause a life-threatening pneumonia termed Legionnaires’ disease. Legionella pneumophila employs the Legionella quorum sensing (Lqs)-LvbR network to regulate virulence and motility, but its role for growth in media is ill-defined. Compared to the parental L. pneumophila strain JR32, a ΔlqsR mutant showed a reduced lag phase at 30°C and reached a higher cell density at 45°C, while the ΔlqsA, ΔlqsS and ΔlqsT mutants exhibited a longer lag phase and reached only a lower cell density. A ΔlvbR mutant resumed growth like the parental strain at 30°C, but exhibited a substantially reduced cell density at 45°C. Thus, LvbR is an important cell density regulator at elevated temperatures. A quantitative analysis of temperature-dependent growth characteristics of environmental and clinical strains revealed that L. pneumophila strains grew in AYE medium after distinct lag phases with similar rates at 30°C, reached different cell densities at the optimal growth temperature of 40°C, and no longer grew at 50°C. Legionella longbeachae reached a rather low cell density at 40°C and did not grow at and beyond 45°C. Genes encoding components of the Lqs-LvbR network were present in the genomes of the environmental and clinical L. pneumophila isolates, and the PlqsR, PlqsA, PlqsS and PlvbR promoters from strain JR32 were active in these strains. Taken together, our results indicate that the Lqs-LvbR network governs the temperature-dependent growth onset and cell density of the L. pneumophila reference strain JR32, and possibly also of environmental and clinical L. pneumophila isolates.

scholarly journals The Legionella Lqs-LvbR regulatory network controls temperature-dependent growth onset and bacterial cell density

2022 ◽  
Author(s):  
Ramon Hochstrasser ◽  
Hubert Hilbi
Keyword(s):  
Quorum Sensing ◽  
Cell Density ◽  
Legionella Pneumophila ◽  
Elevated Temperatures ◽  
Severe Pneumonia ◽  
Water Systems ◽  
Lag Phase ◽  
Temperature Dependent ◽  
Legionnaires Disease ◽  
Dependent Growth

Legionella species are facultative intracellular pathogens, which cause a life-threatening pneumonia termed Legionnaires’ disease. Legionella pneumophila employs the Legionella quorum sensing (Lqs)-LvbR network to regulate virulence and motility, but its role for growth in media is ill-defined. Here we report that compared to the parental L. pneumophila strain JR32, a Δ lqsR mutant showed a reduced lag phase at 30°C and reached a higher cell density at 45°C, while the Δ lqsA , Δ lqsS and Δ lqsT mutants showed a longer lag phase and reached only a lower cell density. A Δ lvbR mutant resumed growth like the parental strain at 30°C, but exhibited a substantially reduced cell density at 45°C. Thus, LvbR is an important cell density regulator at elevated temperatures. Environmental and clinical L. pneumophila strains grew in AYE medium after distinct lag phases with similar rates at 30°C, reached different cell densities at the optimal growth temperature of 40°C, and no longer grew at 50°C. Legionella longbeachae reached a rather low cell density at 40°C and did not grow at and beyond 45°C. Genes encoding components of the Lqs-LvbR network were present in the genomes of the environmental and clinical L. pneumophila isolates, and upon growth at 30°C or 45°C the P lqsR , P lqsA , P lqsS and P lvbR promoters from strain JR32 were expressed in these strains with distinct patterns. Taken together, our results indicate that the Lqs-LvbR network governs the temperature-dependent growth onset and cell density of the L. pneumophila reference strain JR32, and possibly also of environmental and clinical L. pneumophila isolates. Importance Environmental bacteria of the genus Legionella are the causative agents of the severe pneumonia Legionnaires’ disease, the incidence of which is worldwide on the rise. Legionella pneumophila and Legionella longbeachae are the clinically most relevant species. The opportunistic pathogens are inhaled through contaminated aerosols and replicate in human lung macrophages with a similar mechanism as in their natural hosts, free-living amoebae. Given their prevalence in natural and technical water systems, an efficient control of Legionella spp. by physical, chemical or biological means will reduce the incidence of Legionnaires’ disease. Here we show that the Legionella quorum sensing (Lqs) system and the pleiotropic transcription factor LvbR govern the temperature-dependent growth onset and cell density of bacterial cultures. Hence, the growth of L. pneumophila in water systems is not only determined by the temperature and nutrient availability, but also by quorum sensing, i.e., density- and signaling molecule-dependent gene regulation.

Density-Dependent Growth Adaptation Kinetics in 3T 3 Cell Populations Following Sudden [Ca2+] and Temperature Changes. A Comparison with SV40-3T3 Cells

1979 ◽  
Vol 34 (3-4) ◽  
pp. 279-283 ◽  
Author(s):  
Jürgen van der Bosch ◽  
Ilse Sommer ◽  
Heinz Maier ◽  
Willy Rahmig
Keyword(s):  
Growth Rate ◽  
Cell Density ◽  
Cell Number ◽  
Cell Populations ◽  
3T3 Cells ◽  
Density Dependent ◽  
Cell Densities ◽  
Dependent Growth ◽  
Growth Adaptation ◽  
Stationary Cell

Abstract Lowered extracellular [Ca2+] causes low growth rates and low stationary cell densities in 3T3 cell cultures as compared to physiological [Ca2+]. Under otherwise constant conditions the extra­ cellular [Ca2+] determines a stable stationary cell density, which can be readied by increase of net cell number or decrease of net cell number, depending on cell density at the time of [Ca2+] adjustment. SV40-3T3 cells do not show this [Ca2+] dependency. At 39 °C 3T3 and SV40-3T3 cell populations show an increased growth rate at low cell densities as compared to cell populations at 35 °C. Approaching the stationary density the growth rate of both cell sorts is reduced faster at 39 °C than at 35 °C, leading to lower stationary cell densities at 39 °C than at 35 °C. A temperature change from 39 °C to 35 °C or in the opposite direction can affect the stationary cell density of 3T3 cell populations only if applied before reduction of growth rate by density-dependent growth-inhibiting principles has taken place.

scholarly journals Temperature-Dependent Growth Modeling of Environmental and Clinical Legionella pneumophila Multilocus Variable-Number Tandem-Repeat Analysis (MLVA) Genotypes

2017 ◽  
Vol 83 (8) ◽  
Author(s):  
Yehonatan Sharaby ◽  
Sarah Rodríguez-Martínez ◽  
Olga Oks ◽  
Marina Pecellin ◽  
Hila Mizrahi ◽  
...  
Keyword(s):  
Drinking Water ◽  
Growth Kinetics ◽  
Legionella Pneumophila ◽  
Variable Number Tandem Repeat ◽  
Ecological Niches ◽  
Water Network ◽  
Temperature Dependent ◽  
Content Type ◽  
Dependent Growth ◽  
Kinetics Of

ABSTRACT Legionella pneumophila causes waterborne infections resulting in severe pneumonia. High-resolution genotyping of L. pneumophila isolates can be achieved by multiple-locus variable-number tandem-repeat analysis (MLVA). Recently, we found that different MLVA genotypes of L. pneumophila dominated different sites in a small drinking-water network, with a genotype-related temperature and abundance regime. The present study focuses on understanding the temperature-dependent growth kinetics of the genotypes that dominated the water network. Our aim was to model mathematically the influence of temperature on the growth kinetics of different environmental and clinical L. pneumophila genotypes and to compare it with the influence of their ecological niches. Environmental strains showed a distinct temperature preference, with significant differences among the growth kinetics of the three studied genotypes (Gt4, Gt6, and Gt15). Gt4 strains exhibited superior growth at lower temperatures (25 and 30°C), while Gt15 strains appeared to be best adapted to relatively higher temperatures (42 and 45°C). The temperature-dependent growth traits of the environmental genotypes were consistent with their distribution and temperature preferences in the water network. Clinical isolates exhibited significantly higher growth rates and reached higher maximal cell densities at 37°C and 42°C than the environmental strains. Further research on the growth preferences of L. pneumophila clinical and environmental genotypes will result in a better understanding of their ecological niches in drinking-water systems as well as in the human body. IMPORTANCE Legionella pneumophila is a waterborne pathogen that threatens humans in developed countries. The bacteria inhabit natural and man-made freshwater environments. Here we demonstrate that different environmental L. pneumophila genotypes have different temperature-dependent growth kinetics. Moreover, Legionella strains that belong to the same species but were isolated from environmental and clinical sources possess adaptations for growth at different temperatures. These growth preferences may influence the bacterial colonization at specific ecological niches within the drinking-water network. Adaptations for growth at human body temperatures may facilitate the abilities of some L. pneumophila strains to infect and cause illness in humans. Our findings may be used as a tool to improve Legionella monitoring in drinking-water networks. Risk assessment models for predicting the risk of legionellosis should take into account not only Legionella concentrations but also the temperature-dependent growth kinetics of the isolates.

scholarly journals Modelling the Efficacy of Febrile Heating in Infected Endotherms

2021 ◽  
Vol 9 ◽  
Author(s):  
Gregory Lewis ◽  
Michael B. Bonsall
Keyword(s):  
Body Temperature ◽  
Antimicrobial Effect ◽  
Optimal Growth ◽  
Absolute Temperature ◽  
Continuous Heating ◽  
Temperature Dependent ◽  
Simple Physical Model ◽  
Direct Growth ◽  
Dependent Growth ◽  
Body Temperature Increase

Fever is a response to infection characterised by an increase in body temperature. The adaptive value of this body temperature increase for endotherms is unclear, given the relatively small absolute temperature increases associated with endotherm fever, its substantial metabolic costs, and the plausibility for pathogens to adapt to higher temperatures. We consider three thermal mechanisms for fever's antimicrobial effect: (1) direct growth inhibition by elevating temperature above the pathogens optimal growth temperature; (2) further differentiating the host body from the wider environment; and (3) through increasing thermal instability of the pathogen environment. We assess these by modelling their effects pathogen on temperature dependent growth, finding thermal effects can vary from highly to minimally effective depending on pathogen species. We also find, depending on the specification of a simple physical model, intermittent heating can inhibit pathogen growth more effectively than continuous heating with an energy constraint.

scholarly journals In vivo transcriptome analysis provides insights into host-dependent expression of virulence factors by Yersinia entomophaga MH96, during infection of Galleria mellonella

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Amber R Paulson ◽  
Maureen O’Callaghan ◽  
Xue-Xian Zhang ◽  
Paul B Rainey ◽  
Mark R H Hurst
Keyword(s):  
Galleria Mellonella ◽  
Functional Enrichment ◽  
Natural Environments ◽  
Insecticidal Toxin ◽  
Heat Stable ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Cell Densities

Abstract The function of microbes can be inferred from knowledge of genes specifically expressed in natural environments. Here, we report the in vivo transcriptome of the entomopathogenic bacterium Yersinia entomophaga MH96, captured during initial, septicemic, and pre-cadaveric stages of intrahemocoelic infection in Galleria mellonella. A total of 1285 genes were significantly upregulated by MH96 during infection; 829 genes responded to in vivo conditions during at least one stage of infection, 289 responded during two stages of infection, and 167 transcripts responded throughout all three stages of infection compared to in vitro conditions at equivalent cell densities. Genes upregulated during the earliest infection stage included components of the insecticidal toxin complex Yen-TC (chi1, chi2, and yenC1), genes for rearrangement hotspot element containing protein yenC3, cytolethal distending toxin cdtAB, and vegetative insecticidal toxin vip2. Genes more highly expressed throughout the infection cycle included the putative heat-stable enterotoxin yenT and three adhesins (usher-chaperone fimbria, filamentous hemagglutinin, and an AidA-like secreted adhesin). Clustering and functional enrichment of gene expression data also revealed expression of genes encoding type III and VI secretion system-associated effectors. Together these data provide insight into the pathobiology of MH96 and serve as an important resource supporting efforts to identify novel insecticidal agents.

scholarly journals Physical networks from entropy-driven non-covalent interactions

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anthony C. Yu ◽  
Huada Lian ◽  
Xian Kong ◽  
Hector Lopez Hernandez ◽  
Jian Qin ◽  
...  
Keyword(s):  
Elevated Temperatures ◽  
Dissociation Rate ◽  
Mathematical Framework ◽  
Temperature Dependent ◽  
Nanoparticle Interactions ◽  
Temperature Superposition ◽  
Non Covalent Interactions ◽  
Time Temperature Superposition ◽  
Covalent Interactions ◽  
Physical Crosslinking

AbstractPhysical networks typically employ enthalpy-dominated crosslinking interactions that become more dynamic at elevated temperatures, leading to network softening. Moreover, standard mathematical frameworks such as time-temperature superposition assume network softening and faster dynamics at elevated temperatures. Yet, deriving a mathematical framework connecting the crosslinking thermodynamics to the temperature-dependent viscoelasticity of physical networks suggests the possibility for entropy-driven crosslinking interactions to provide alternative temperature dependencies. This framework illustrates that temperature negligibly affects crosslink density in reported systems, but drastically influences crosslink dynamics. While the dissociation rate of enthalpy-driven crosslinks is accelerated at elevated temperatures, the dissociation rate of entropy-driven crosslinks is negligibly affected or even slowed under these conditions. Here we report an entropy-driven physical network based on polymer-nanoparticle interactions that exhibits mechanical properties that are invariant with temperature. These studies provide a foundation for designing and characterizing entropy-driven physical crosslinking motifs and demonstrate how these physical networks access thermal properties that are not observed in current physical networks.

scholarly journals The Early Transcriptional Response of Human Granulocytes to Infection with Candida albicans Is Not Essential for Killing but Reflects Cellular Communications

2006 ◽  
Vol 75 (3) ◽  
pp. 1493-1501 ◽  
Author(s):  
Chantal Fradin ◽  
Abigail L. Mavor ◽  
Günther Weindl ◽  
Martin Schaller ◽  
Karin Hanke ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Mononuclear Cells ◽  
De Novo ◽  
Transcriptional Response ◽  
Mononuclear Leukocytes ◽  
General Notion ◽  
Genes Encoding ◽  
Global Transcriptional Profile ◽  
Life Threatening ◽  
Systemic Infections

ABSTRACT Candida albicans is a polymorphic opportunistic fungus that can cause life-threatening systemic infections following hematogenous dissemination in patients susceptible to nosocomial infection. Neutrophils form part of the innate immune response, which is the first line of defense against microbes and is particularly important in C. albicans infections. To compare the transcriptional response of leukocytes exposed to C. albicans, we investigated the expression of key cytokine genes in polymorphonuclear and mononuclear leukocytes after incubation with C. albicans for 1 h. Isolated mononuclear cells expressed high levels of genes encoding proinflammatory signaling molecules, whereas neutrophils exhibited much lower levels, similar to those observed in whole blood. The global transcriptional profile of neutrophils was examined by using an immunology-biased human microarray to determine whether different morphological forms or the viability of C. albicans altered the transcriptome. Hyphal cells appeared to have the broadest effect, although the most strongly induced genes were regulated independently of morphology or viability. These genes were involved in proinflammatory cell-cell signaling, cell signal transduction, and cell growth. Generally, genes encoding known components of neutrophil granules showed no upregulation at this time point; however, lactoferrin, a well-known candidacidal peptide, was secreted by neutrophils. Addition to inhibitors of RNA or protein de novo synthesis did not influence the killing activity within 30 min. These results support the general notion that neutrophils do not require gene transcription to mount an immediate and direct attack against microbes. However, neutrophils exposed to C. albicans express genes involved in communication with other immune cells.

Conductivity-Temperature Dependent Studies on MG49 Doped Lithium Triflate Salt

2015 ◽  
Vol 1107 ◽  
pp. 181-186
Author(s):  
Zaidatul Salwa Mahmud ◽  
N.H.M. Zaki ◽  
R. Zakaria ◽  
Mohamad Faizul Yahya ◽  
Ab Malik Marwan Ali
Keyword(s):  
Ionic Conductivity ◽  
Elevated Temperatures ◽  
Ethylene Carbonate ◽  
Ionic Mobility ◽  
Ionic Conductors ◽  
X Ray Diffraction ◽  
Temperature Dependent ◽  
Lithium Triflate ◽  
Mobility Of Charge Carriers ◽  
Solution Cast

This paper reports on the conductivity-temperature studies of gel polymer electrolytes (GPEs) based on 49% poly (methyl methacrylate) grafted-natural rubber (MG49) doped with lithium triflate salt (LiTf) and plasticized with ethylene carbonate (EC). The GPE films are prepared by solution cast technique. The X-ray diffraction (XRD) studies reveal the polymer electrolyte systems are amorphous. AC impedance spectroscopy is carried out in the temperature range between 303 and 373 K. The magnitudes of conductivity observed are strongly dependent on salt concentration and temperature. The high ionic conductivity at elevated temperatures of GPE is attributed to the high ionic mobility of charge carriers. The ionic migration is seen to follow the VTF behavior and approaches to Arrhenius rule at high and low at temperature. Ionic conductivity relaxation appears to be a characteristic of the ionic polarization and the modulus formalism studies confirmed the GPEs in the present investigation are ionic conductors.

scholarly journals Membrane-Based Separation of Phenol/Water Mixtures Using Ionically and Covalently Cross-Linked Ethylene-Methacrylic Acid Copolymers

2008 ◽  
Vol 2008 ◽  
pp. 1-12 ◽  
Author(s):  
Alexander Mixa ◽  
Claudia Staudt
Keyword(s):  
Methacrylic Acid ◽  
Elevated Temperatures ◽  
Type Equation ◽  
Enrichment Factors ◽  
Cross Linking ◽  
Temperature Dependent ◽  
Monomer Content ◽  
Phenol Water ◽  
Feed Temperature ◽  
Acetyl Acetonate

Membrane-based separation of phenol/water mixtures with concentrations of phenol between 3 wt% and 8 wt% in the feed has been performed with nonmodified as well as cross-linked ethylene-methacrylic acid (E-MAA) copolymers with different amounts of methacrylic acid. As cross-linking agents, aluminium acetyl acetonate, which leads to ionically cross-linked membranes, and 2,3,5,6-tetramethyl-1,4-phenylene diamine and glycerine digycidether, leading to covalently cross-linked membranes, have been used. Generally, it was found that with increasing phenol content in the feed, the total flux is increasing whereas the enrichment factor is decreasing. Using nonmodified membranes with higher methacrylic acid monomer content in the polymer, lower fluxes and higher enrichment factors were observed. Investigation of different cross-linked membranes showed that with high phenol concentration in the feed, ionic cross-linking seems to be very promising. Furthermore, variation of feed temperature shows that ionically cross-linked membranes reached higher fluxes as well as higher enrichment factors at elevated temperatures. The temperature-dependent data were fitted based on an Arrhenius-type equation, and activation energies for the permeation of phenol and water through the membrane were calculated.

scholarly journals Platform Engineering of Corynebacterium glutamicum with Reduced Pyruvate Dehydrogenase Complex Activity for Improved Production of l-Lysine, l-Valine, and 2-Ketoisovalerate

2013 ◽  
Vol 79 (18) ◽  
pp. 5566-5575 ◽  
Author(s):  
Jens Buchholz ◽  
Andreas Schwentner ◽  
Britta Brunnenkan ◽  
Christina Gabris ◽  
Simon Grimm ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Fed Batch ◽  
Complex Activity ◽  
Gene Encoding ◽  
Content Type ◽  
Genes Encoding ◽  
Cell Densities ◽  
Dehydrogenase Complex

ABSTRACTExchange of the nativeCorynebacterium glutamicumpromoter of theaceEgene, encoding the E1p subunit of the pyruvate dehydrogenase complex (PDHC), with mutateddapApromoter variants led to a series ofC. glutamicumstrains with gradually reduced growth rates and PDHC activities. Upon overexpression of thel-valine biosynthetic genesilvBNCE, all strains producedl-valine. Among these strains,C. glutamicum aceEA16 (pJC4ilvBNCE) showed the highest biomass and product yields, and thus it was further improved by additional deletion of thepqoandppcgenes, encoding pyruvate:quinone oxidoreductase and phosphoenolpyruvate carboxylase, respectively. In fed-batch fermentations at high cell densities,C. glutamicum aceEA16 Δpqo Δppc(pJC4ilvBNCE) produced up to 738 mM (i.e., 86.5 g/liter)l-valine with an overall yield (YP/S) of 0.36 mol per mol of glucose and a volumetric productivity (QP) of 13.6 mM per h [1.6 g/(liter × h)]. Additional inactivation of the transaminase B gene (ilvE) and overexpression ofilvBNCDinstead ofilvBNCEtransformed thel-valine-producing strain into a 2-ketoisovalerate producer, excreting up to 303 mM (35 g/liter) 2-ketoisovalerate with aYP/Sof 0.24 mol per mol of glucose and aQPof 6.9 mM per h [0.8 g/(liter × h)]. The replacement of theaceEpromoter by thedapA-A16 promoter in the twoC. glutamicuml-lysine producers DM1800 and DM1933 improved the production by 100% and 44%, respectively. These results demonstrate thatC. glutamicumstrains with reduced PDHC activity are an excellent platform for the production of pyruvate-derived products.

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