[Formula: see text]-linear resistivity from magneto-elastic scattering: Application to PdCrO2.

An electronic solid with itinerant carriers and localized magnetic moments represents a paradigmatic strongly correlated system. The electrical transport properties associated with the itinerant carriers, as they scatter off these local moments, have been scrutinized across a number of materials. Here, we analyze the transport characteristics associated with ultraclean PdCrO[Formula: see text]-a quasi-two-dimensional material consisting of alternating layers of itinerant Pd-electrons and Mott-insulating CrO[Formula: see text] layers-which shows a pronounced regime of T-linear resistivity over a wide range of intermediate temperatures. By contrasting these observations to the transport properties in a closely related material PdCoO[Formula: see text], where the CoO[Formula: see text] layers are band-insulators, we can rule out the traditional electron-phonon interactions as being responsible for this interesting regime. We propose a previously ignored electron-magneto-elastic interaction between the Pd-electrons, the Cr local moments and an out-of-plane phonon as the main scattering mechanism that leads to the significant enhancement of resistivity and a T-linear regime in PdCrO[Formula: see text] at temperatures far in excess of the magnetic ordering temperature. We suggest a number of future experiments to confirm this picture in PdCrO[Formula: see text] as well as other layered metallic/Mott-insulating materials.

Enhancer-instructed epigenetic landscape and chromatin compartmentalization dictate a primary antibody repertoire protective against specific bacterial pathogens.

Antigen receptor loci are organized into variable (V), diversity (D) and joining (J) gene segments that rearrange to generate antigen receptor repertoires. Here, we identified an enhancer (E34) in the murine immunoglobulin kappa (Igk) locus that instructed rearrangement of Vκ genes located in a sub-topologically associating domain, including a Vκ gene encoding for antibodies targeting bacterial phosphorylcholine. We show that E34 instructs the nuclear repositioning of the E34 sub-topologically associating domain from a recombination-repressive compartment to a recombination-permissive compartment that is marked by equivalent activating histone modifications. Finally, we found that E34-instructed Vκ-Jκ rearrangement was essential to combat Streptococcus pneumoniae but not methicillin-resistant Staphylococcus aureus or influenza infections. We propose that the merging of Vκ genes with Jκ elements is instructed by one-dimensional epigenetic information imposed by enhancers across Vκ and Jκ genomic regions. The data also reveal how enhancers generate distinct antibody repertoires that provide protection against lethal bacterial infection.

Pressure-Surface Tension-Temperature Equation of State for n-Alkanes.

Herein, the geometric similitude concept is applied to propose a cubic equation that relates surface tension, saturation pressure, and temperature for n-alkanes. The input properties for each fluid are the molecular mass, pressure, temperature, and compressibility factor at the critical point. The model is applied to temperatures below 0.93·T c (critical point temperature). A total of 2429 surface tension values have been selected for 32 n-alkanes. The parameters of the model have been obtained with a fit of the surface tension values for 19 pure n-alkanes that are randomly chosen. Then, it is tested for the other 13 pure n-alkanes and used to predict the surface tension for 11 binary and 4 ternary mixtures. These predictions are compared with the reported experimental data. For pure n-alkanes, the overall absolute average deviation is 2.4%, including the correlation and testing sets. No additional adjustable coefficients are used for mixtures, yielding an overall absolute average deviation of 2.98% for the binary systems and 7.97% for the ternary ones. The results show that the model is accurate enough for predictions and that the highest deviations are due to the lack of agreement in the values of surface tension of pure fluids obtained from different sources.

Evaluation of IL-17D in Host Immunity to Group A Streptococcus Infection.

IL-17D is a cytokine that belongs to the IL-17 family and is conserved in vertebrates and invertebrates. In contrast to IL-17A and IL-17F, which are expressed in Th17 cells, IL-17D is expressed broadly in nonimmune cells. IL-17D can promote immune responses to cancer and viruses in part by inducing chemokines and recruiting innate immune cells such as NK cells. Although bacterial infection can induce IL-17D in fish and invertebrates, the role of mammalian IL-17D in antibacterial immunity has not been established. To determine whether IL-17D has a role in mediating host defense against bacterial infections, we studied i.p. infection by group A Streptococcus (GAS) in wild-type (WT) and Il17d -/- mice. Compared with WT animals, mice deficient in IL-17D experienced decreased survival, had greater weight loss, and showed increased bacterial burden in the kidney and peritoneal cavity following GAS challenge. In WT animals, IL-17D transcript was induced by GAS infection and correlated to increased levels of chemokine CCL2 and greater neutrophil recruitment. Of note, GAS-mediated IL-17D induction in nonimmune cells required live bacteria, suggesting that processes beyond recognition of pathogen-associated molecular patterns were required for IL-17D induction. Based on our results, we propose a model in which nonimmune cells can discriminate between nonviable and viable GAS cells, responding only to the latter by inducing IL-17D.

Dynamic Interactions of Group A Streptococcus with Host Macrophages.

Macrophages play a critical role in Group A Streptococcus (GAS) recognition and the consequent activation of innate immunity and inflammatory responses against the pathogen. In parallel, GAS deploys several strategies for escaping detection and elimination by these efficient phagocytic cells. The events that take place in this GAS-macrophage battleground, the cellular consequences for the pathogen and for the immune cell, and the balance between the magnitude of infection and the efficiency of the host immune response can be investigated with a variety of assays presented in this chapter.

A bacterial gene-drive system efficiently edits and inactivates a high copy number antibiotic resistance locus.

Gene-drive systems in diploid organisms bias the inheritance of one allele over another. CRISPR-based gene-drive expresses a guide RNA (gRNA) into the genome at the site where the gRNA directs Cas9-mediated cleavage. In the presence of Cas9, the gRNA cassette and any linked cargo sequences are copied via homology-directed repair (HDR) onto the homologous chromosome. Here, we develop an analogous CRISPR-based gene-drive system for the bacterium Escherichia coli that efficiently copies a gRNA cassette and adjacent cargo flanked with sequences homologous to the targeted gRNA/Cas9 cleavage site. This "pro-active" genetic system (Pro-AG) functionally inactivates an antibiotic resistance marker on a high copy number plasmid with ~ 100-fold greater efficiency than control CRISPR-based methods, suggesting an amplifying positive feedback loop due to increasing gRNA dosage. Pro-AG can likewise effectively edit large plasmids or single-copy genomic targets or introduce functional genes, foreshadowing potential applications to biotechnology or biomedicine.

Functional and Proteomic Analysis of Streptococcus pyogenes Virulence Upon Loss of Its Native Cas9 Nuclease.

The public health impact of Streptococcus pyogenes (group A Streptococcus, GAS) as a top 10 cause of infection-related mortality in humans contrasts with its benefit to biotechnology as the main natural source of Cas9 nuclease, the key component of the revolutionary CRISPR-Cas9 gene editing platform. Despite widespread knowledge acquired in the last decade on the molecular mechanisms by which GAS Cas9 achieves precise DNA targeting, the functions of Cas9 in the biology and pathogenesis of its native organism remain unknown. In this study, we generated an isogenic serotype M1 GAS mutant deficient in Cas9 protein and compared its behavior and phenotypes to the wild-type parent strain. Absence of Cas9 was linked to reduced GAS epithelial cell adherence, reduced growth in human whole blood ex vivo, and attenuation of virulence in a murine necrotizing skin infection model. Virulence defects of the GAS Δcas9 strain were explored through quantitative proteomic analysis, revealing a significant reduction in the abundance of key GAS virulence determinants. Similarly, deletion of cas9 affected the expression of several known virulence regulatory proteins, indicating that Cas9 impacts the global architecture of GAS gene regulation.

A Novel Redox-Sensing Histidine Kinase That Controls Carbon Catabolite Repression in Azoarcus sp. CIB.

We have identified and characterized the AccS multidomain sensor kinase that mediates the activation of the AccR master regulator involved in carbon catabolite repression (CCR) of the anaerobic catabolism of aromatic compounds in Azoarcus sp. CIB. A truncated AccS protein that contains only the soluble C-terminal autokinase module (AccS') accounts for the succinate-dependent CCR control. In vitro assays with purified AccS' revealed its autophosphorylation, phosphotransfer from AccS'∼P to the Asp60 residue of AccR, and the phosphatase activity toward its phosphorylated response regulator, indicating that the equilibrium between the kinase and phosphatase activities of AccS' may control the phosphorylation state of the AccR transcriptional regulator. Oxidized quinones, e.g., ubiquinone 0 and menadione, switched the AccS' autokinase activity off, and three conserved Cys residues, which are not essential for catalysis, are involved in such inhibition. Thiol oxidation by quinones caused a change in the oligomeric state of the AccS' dimer resulting in the formation of an inactive monomer. This thiol-based redox switch is tuned by the cellular energy state, which can change depending on the carbon source that the cells are using. This work expands the functional diversity of redox-sensitive sensor kinases, showing that they can control new bacterial processes such as CCR of the anaerobic catabolism of aromatic compounds. The AccSR two-component system is conserved in the genomes of some betaproteobacteria, where it might play a more general role in controlling the global metabolic state according to carbon availability.IMPORTANCE Two-component signal transduction systems comprise a sensor histidine kinase and its cognate response regulator, and some have evolved to sense and convert redox signals into regulatory outputs that allow bacteria to adapt to the altered redox environment. The work presented here expands knowledge of the functional diversity of redox-sensing kinases to control carbon catabolite repression (CCR), a phenomenon that allows the selective assimilation of a preferred compound among a mixture of several carbon sources. The newly characterized AccS sensor kinase is responsible for the phosphorylation and activation of the AccR master regulator involved in CCR of the anaerobic degradation of aromatic compounds in the betaproteobacterium Azoarcus sp. CIB. AccS seems to have a thiol-based redox switch that is modulated by the redox state of the quinone pool. The AccSR system is conserved in several betaproteobacteria, where it might play a more general role controlling their global metabolic state.

Trichomonas vaginalis Induces NLRP3 Inflammasome Activation and Pyroptotic Cell Death in Human Macrophages.

Trichomonas vaginalis is a sexually transmitted, eukaryotic parasite that causes trichomoniasis, the most common nonviral, sexually transmitted disease in the USA and worldwide. Little is known about the molecular mechanisms involved in the host immune response to this widespread parasite. Here we report that T. vaginalis induces NLRP3 inflammasome activation in human macrophages, leading to caspase-1 activation and the processing of pro-IL-1ß to the mature and bioactive form of the cytokine. Using inhibitor-based approaches, we show that NLRP3 activation by T. vaginalis involves host cell detection of extracellular ATP via P2X7 receptors and potassium efflux. In addition, our data reveal that T. vaginalis inflammasome activation induces macrophage inflammatory cell death by pyroptosis, known to occur via caspase-1 cleavage of the gasdermin D protein, which assembles to form pores in the host cell membrane. We found that T. vaginalis-induced cytolysis of macrophages is attenuated in gasdermin D knockout cells. Lastly, in a murine challenge model, we detected IL-1ß production in vaginal fluids in response to T. vaginalis infection in vivo. Together, our findings mechanistically dissect how T. vaginalis contributes to the production of the proinflammatory IL-1ß cytokine and uncover pyroptosis as a mechanism by which the parasite can trigger host macrophage cell death.

Density-functional fluctuation theory of crowds.

A primary goal of collective population behavior studies is to determine the rules governing crowd distributions in order to predict future behaviors in new environments. Current top-down modeling approaches describe, instead of predict, specific emergent behaviors, whereas bottom-up approaches must postulate, instead of directly determine, rules for individual behaviors. Here, we employ classical density functional theory (DFT) to quantify, directly from observations of local crowd density, the rules that predict mass behaviors under new circumstances. To demonstrate our theory-based, data-driven approach, we use a model crowd consisting of walking fruit flies and extract two functions that separately describe spatial and social preferences. The resulting theory accurately predicts experimental fly distributions in new environments and provides quantification of the crowd "mood". Should this approach generalize beyond milling crowds, it may find powerful applications in fields ranging from spatial ecology and active matter to demography and economics.

Staphylococcus aureus Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models.

Staphylococcus aureus produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that S. aureus-derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from S. aureus grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified S. aureus MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of S. aureus to killing by whole blood or purified human neutrophils ex vivo and increased S. aureus survival in vivo. Finally, immunization of mice with S. aureus-derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic S. aureus infection. Collectively, our results suggest S. aureus MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection.

Group A Streptococcus encounters with host macrophages.

Group A Streptococcus (GAS) is a leading human bacterial pathogen with diverse clinical manifestations. Macrophages constitute a critical first line of host defense against GAS infection, using numerous surface and intracellular receptors such as Toll-like receptors and inflammasomes for pathogen recognition and activation of inflammatory signaling pathways. Depending on the intensity of the GAS infection, activation of these signaling cascades may provide a beneficial early alarm for effective immune clearance, or conversely, may cause hyperinflammation and tissue injury during severe invasive infection. Although traditionally considered an extracellular pathogen, GAS can invade and replicate within macrophages using specific molecular mechanisms to resist phagolysosomal and xenophagic killing. Unraveling GAS-macrophage encounters may reveal new treatment options for this leading agent of infection-associated mortality. [Formula: see text].

Group A streptococcal M protein activates the NLRP3 inflammasome.

Group A Streptococcus (GAS) is among the top ten causes of infection-related mortality in humans. M protein is the most abundant GAS surface protein, and M1 serotype GAS strains are associated with invasive infections, including necrotizing fasciitis and toxic shock syndrome. Here, we report that released, soluble M1 protein triggers programmed cell death in macrophages (Mϕ). M1 served as a second signal for caspase-1-dependent NLRP3 inflammasome activation, inducing maturation and release of proinflammatory cytokine interleukin-1ß (IL-1ß) and macrophage pyroptosis. The structurally dynamic B-repeat domain of M1 was critical for inflammasome activation, which involved K+ efflux and M1 protein internalization by clathrin-mediated endocytosis. Mouse intraperitoneal challenge showed that soluble M1 was sufficient and specific for IL-1ß activation, which may represent an early warning to activate host immunity against the pathogen. Conversely, in systemic infection, hyperinflammation associated with M1-mediated pyroptosis and IL-1ß release could aggravate tissue injury.

Serine-Aspartate Repeat Protein D Increases Staphylococcus aureus Virulence and Survival in Blood.

Staphylococcus aureus expresses a panel of cell wall-anchored adhesins, including proteins belonging to the microbial surface components recognizing adhesive matrix molecule (MSCRAMM) family, exemplified by the serine-aspartate repeat protein D (SdrD), which serve key roles in colonization and infection. Deletion of sdrD from S. aureus subsp. aureus strain NCTC8325-4 attenuated bacterial survival in human whole blood ex vivo, which was associated with increased killing by human neutrophils. Remarkably, SdrD was able to inhibit innate immune-mediated bacterial killing independently of other S. aureus proteins, since addition of recombinant SdrD protein and heterologous expression of SdrD in Lactococcus lactis promoted bacterial survival in human blood. SdrD contributes to bacterial virulence in vivo, since fewer S. aureus subsp. aureus NCTC8325-4 ΔsdrD bacteria than bacteria of the parent strain were recovered from blood and several organs using a murine intravenous infection model. Collectively, our findings reveal a new property of SdrD as an important key contributor to S. aureus survival and the ability to escape the innate immune system in blood.

Coiled-coil destabilizing residues in the group A Streptococcus M1 protein are required for functional interaction.

The sequences of M proteins, the major surface-associated virulence factors of the widespread bacterial pathogen group A Streptococcus, are antigenically variable but have in common a strong propensity to form coiled coils. Paradoxically, these sequences are also replete with coiled-coil destabilizing residues. These features are evident in the irregular coiled-coil structure and thermal instability of M proteins. We present an explanation for this paradox through studies of the B repeats of the medically important M1 protein. The B repeats are required for interaction of M1 with fibrinogen (Fg) and consequent proinflammatory activation. The B repeats sample multiple conformations, including intrinsically disordered, dissociated, as well as two alternate coiled-coil conformations: a Fg-nonbinding register 1 and a Fg-binding register 2. Stabilization of M1 in the Fg-nonbinding register 1 resulted in attenuation of Fg binding as expected, but counterintuitively, so did stabilization in the Fg-binding register 2. Strikingly, these register-stabilized M1 proteins gained the ability to bind Fg when they were destabilized by a chaotrope. These results indicate that M1 stability is antithetical to Fg interaction and that M1 conformational dynamics, as specified by destabilizing residues, are essential for interaction. A "capture-and-collapse" model of association accounts for these observations, in which M1 captures Fg through a dynamic conformation and then collapses into a register 2-coiled coil as a result of stabilization provided by binding energy. Our results support the general conclusion that destabilizing residues are evolutionarily conserved in M proteins to enable functional interactions necessary for pathogenesis.

Thermoresponsive microgels at the air-water interface: the impact of the swelling state on interfacial conformation.

Poly(N-vinylcaprolactam) (PVCL) is a new temperature-responsive type of polymer microgel with improved biocompatibility as compared to more commonly used poly(N-isopropylacrylamide) (PNIPAM). Both polymers swell at low temperatures and collapse at high ones, showing a volume phase transition temperature (VPTT) around the physiological temperature. Exploring the interfacial characteristics of thermoresponsive microgels is important due to their potential application in emulsion based systems with tailored stabilities and controlled degradation profiles. In this work, we study the properties of charged PVCL particles at the air-water interface by a combination of adsorption, dilatational rheology and Langmuir monolayers. Although PVCL particles adsorb spontaneously at the air-water interface in both, swollen and collapsed conformations, the interfacial properties show significant differences depending on the swelling state. In particular, the total amount of adsorbed microgels and the rigidity of the monolayer increase as the temperature increases above the VPTT, which is connected to the more compact morphology of the microgels in this regime. Dilatational rheology data show the formation of a very loose adsorbed layer with low cohesivity. In addition, collapsed microgels yield a continuous increase of the surface pressure, whereas swollen microgels show a phase transition at intermediate compressions caused by the deformation of the loose external polymer shell of the particles. We also provide a qualitative interpretation for the surface pressure behavior in terms of microgel-microgel effective pair potentials, and correlate our experimental findings to recent rescaling models that take into account the importance of the internal polymer degrees of freedom in the rearrangement of the conformation of the microgel particles at the interface.

The cacao pathogen Moniliophthora roreri (Marasmiaceae) possesses biallelic A and B mating loci but reproduces clonally.

The cacao pathogen Moniliophthora roreri belongs to the mushroom-forming family Marasmiaceae, but it has never been observed to produce a fruiting body, which calls to question its capacity for sexual reproduction. In this study, we identified potential A (HD1 and HD2) and B (pheromone precursors and pheromone receptors) mating genes in M. roreri. A PCR-based method was subsequently devised to determine the mating type for a set of 47 isolates from across the geographic range of the fungus. We developed and generated an 11-marker microsatellite set and conducted association and linkage disequilibrium (standardized index of association, IA(s)) analyses. We also performed an ancestral reconstruction analysis to show that the ancestor of M. roreri is predicted to be heterothallic and tetrapolar, which together with sliding window analyses support that the A and B mating loci are likely unlinked and follow a tetrapolar organization within the genome. The A locus is composed of a pair of HD1 and HD2 genes, whereas the B locus consists of a paired pheromone precursor, Mr_Ph4, and receptor, STE3_Mr4. Two A and B alleles but only two mating types were identified. Association analyses divided isolates into two well-defined genetically distinct groups that correlate with their mating type; IA(s) values show high linkage disequilibrium as is expected in clonal reproduction. Interestingly, both mating types were found in South American isolates but only one mating type was found in Central American isolates, supporting a prior hypothesis of clonal dissemination throughout Central America after a single or very few introductions of the fungus from South America.

Evaluating the influence of physical, economic and managerial factors on sheet erosion in rangelands of SW Spain by performing a sensitivity analysis on an integrated dynamic model.

An integrated dynamic model was used to evaluate the influence of climatic, soil, pastoral, economic and managerial factors on sheet erosion in rangelands of SW Spain (dehesas). This was achieved by means of a variance-based sensitivity analysis. Topsoil erodibility, climate change and a combined factor related to soil water storage capacity and the pasture production function were the factors which influenced water erosion the most. Of them, climate change is the main source of uncertainty, though in this study it caused a reduction in the mean and the variance of long-term erosion rates. The economic and managerial factors showed scant influence on soil erosion, meaning that it is unlikely to find such influence in the study area for the time being. This is because the low profitability of the livestock business maintains stocking rates at low levels. However, the potential impact of livestock, through which economic and managerial factors affect soil erosion, proved to be greater in absolute value than the impact of climate change. Therefore, if changes in some economic or managerial factors led to higher stocking rates in the future, significant increases in erosion rates would be expected.

Anatomical variations of the celiac trunk: cadaveric and radiological study.

PURPOSE: The celiac trunk (CT) is a vascular structure of the upper abdomen which gives off the left gastric artery (LGA), the splenic artery and the common hepatic artery. This study aims to compare the vascular patterns of the CT of two different samples (cadaveric and radiological) and to propose a simple classification of CT variations based on previous studies and our results. MATERIALS AND METHODS: To perform this study we examined 43 adult cadavers, 24 males and 19 females, ages ranged from 69 to 92. In addition, we analysed 596 MDCT (multidetector computed tomographic) angiography examinations of 430 males and 166 females, ages ranged from 42 to 82. RESULTS: According to the classification proposed, results were divided into Type I or complete CT (578/639 cases, 90.5 %), Type II or incomplete CT (61/639 cases, 9.5 %), Type III or absence of CT and Type IV or celiacomesenteric trunk with no cases reported. Type I was divided into Type Ia or bifurcated trunk with LGA arising first (368/639 cases, 57.6 %), Type Ib or trifurcated trunk (205/639 cases, 32.1 %) and Type Ic or tetrafurcated trunk with an extra branch (5/639 cases, 0.8 %). Type II included hepatosplenic (29/639 cases, 4.5 %), gastroplenic (32/639, 5 %) and hepatogastric trunks (0/639, 0 %) which represented Types IIa, IIb and IIc respectively. CONCLUSIONS: No significant differences were found between the cadaveric and radiological samples. Gender did not appear to be related to any variability of the structures either. A new, simple and complete classification of the anatomical variations of the CT is proposed.

Bile salts at the air-water interface: adsorption and desorption.

Bile salts (BS) are bio-surfactants which constitute a vital component in the process of fat digestion. Despite the importance of the interfacial properties in their biological role, these have been scarcely studied in the literature. In this work, we present the adsorption-desorption profiles of two BS (NaTC and NaGDC) including dilatational rheology. Findings from this study reveal very different surface properties of NaTC and NaGDC which originate from different complexation properties relevant to the digestion process. Dynamic adsorption curves show higher adsorption rates for NaTC and suggest the existence of various conformational regimes in contrast to NaGDC which presents only one conformational regime. This is corroborated by analysis of the adsorption isotherms and more in detail by the rheological behaviour. Accordingly, the dilatational response at 1Hz displays two maxima of the dilatational modulus for NaTC as a function of bulk concentration, in contrast to NaGDC which displays only one maximum. The desorption profiles reveal that NaTC adopts an irreversibly adsorbed form at high surface coverage whereas NaGDC fully desorbs from the surface within the whole range of concentrations used. Analysis of the adsorption-desorption profiles provides new insight into the surface properties of BS, suggesting a surface complexation of NaTC. This knowledge can be useful since through interfacial engineering we might control the extent of lipolysis providing the basis for the rational design of food products with tailored digestibility.