Pseudomonas aeruginosa Lipid A Structural Variants Induce Altered Immune Responses.

Pseudomonas aeruginosa causes chronic lung infection in cystic fibrosis (CF), resulting in structural lung damage and progressive pulmonary decline. P. aeruginosa in the CF lung undergoes numerous changes, adapting to host-specific airway pressures while establishing chronic infection. P. aeruginosa undergoes lipid A structural modification during CF chronic infection, not seen in any other disease state. Lipid A, the membrane anchor of lipopolysaccharide (i.e., endotoxin), comprises the majority of the outer membrane of Gram-negative bacteria and is a potent toll-like receptor (TLR)4 agonist. The structure of P. aeruginosa lipid A is intimately linked with its recognition by TLR4, and subsequent immune response. Prior work has identified P. aeruginosa strains with altered lipid A structures that arise during chronic CF lung infection; however, the impact of P. aeruginosa lipid A structure on airway disease has not been investigated. Here, we show that P. aeruginosa lipid A lacks PagL-mediated deacylation during human airway infection using a direct-from-sample mass spectrometry approach on human bronchoalveolar lavage fluid. This structure triggers increased pro-inflammatory cytokine production by primary human macrophages. Furthermore, alterations in lipid A 2-hydroxylation impact cytokine response in a site-specific manner, independent of CFTR function. Interestingly, there is a CF-specific reduction in IL-8 secretion within the epithelial-cell compartment that only occurs in CF bronchial epithelial cells when infected with CF-adapted P. aeruginosa that lack PagL-mediated lipid A deacylation. Taken together, we show that P. aeruginosa alters its lipid A structure during acute lung infection and that this lipid A structure induces stronger signaling through TLR4.

Immunometabolic Regulation of Bacterial Infection, Biofilms, and Antibiotic Susceptibility.

BACKGROUND: Upon infection, mucosal tissues activate a brisk inflammatory response to clear the pathogen, i.e., resistance to disease. Resistance to disease is orchestrated by tissue-resident macrophages, which undergo profound metabolic reprogramming after sensing the pathogen. These metabolically activated macrophages release many inflammatory factors, which promote their bactericidal function. However, in immunocompetent individuals, pathogens like Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella evade this type of immunity, generating communities that thrive for the long term. SUMMARY: These organisms develop features that render them less susceptible to eradication, such as biofilms and increased tolerance to antibiotics. Furthermore, after antibiotic therapy withdrawal, "persister" cells rapidly upsurge, triggering inflammatory relapses that worsen host health. How these pathogens persisted in inflamed tissues replete with activated macrophages remains poorly understood. KEY MESSAGES: In this review, we discuss recent findings indicating that the ability of P. aeruginosa, S. aureus, and Salmonella to evolve biofilms and antibiotic tolerance is promoted by the similar metabolic routes that regulate macrophage metabolic reprogramming.

Desperdicio de alimentos en comedores escolares de una región de la zona centro sur de Chile: Percepciones de manipuladoras de alimentos del Programa de Alimentación Escolar

Introducción: Los desperdicios de alimentos en los comedores escolares son un problema que afecta al uso de recursos naturales, humanos y económicos. Objetivo: Identificar las opiniones y percepciones de manipuladoras de alimentos sobre factores relacionados al desperdicio de alimentos y posibles estrategias para disminuirlo en comedores escolares del Programa de Alimentación Escolar de la Junta Nacional de Auxilio Escolar y Becas en Chile. Métodos: Diseño cualitativo mediante técnica de grupos focales, con 33 participantes. Se aplicó un protocolo de investigación cualitativo que incluyó consentimiento informado, caracterización sociodemográfica, pauta de observación y guion con las dimensiones exploradas. Cada grupo contó con moderador experto y un observador. Se grabó y transcribió el audio de cada sesión y se analizó mediante ATLAS. Ti, versión 8. Resultados: Según las manipuladoras, los alimentos menos consumidos por los estudiantes son las legumbres, verduras frescas, productos marinos y algunas salsas. Estiman que el desperdicio de alimentos oscila entre los 25 a 100 Kilos al día. Las mujeres identifican factores claves en el rechazo de los alimentos por parte de los escolares; estética, olor y sabor del plato; repetitividad del menú; y aspectos culturales. Como estrategias para reducir el desperdicio proponen variar el menú, incorporar alimentos conocidos por los escolares, educar a los padres en la importancia de ofrecer alimentos y preparaciones saludables y sostenibles; fomentar la participación de toda la comunidad educativa en el proceso alimentario. Conclusiones: El estudio reveló que el desperdicio de alimentos en los comedores escolares es un problema complejo que requiere una intervención multidimensional que involucre a todos los actores del sistema alimentario escolar.

Introduction: Food waste in school canteens is a problem that affects the use of natural, human and economic resources. Objective: Identify the opinions and perceptions of food handlers about food waste factors and possible strategies to reduce them in school canteens of the School Feeding Programme of the Junta Nacional de Auxilio Escolar y Becas in Chile. Methods: Qualitative design using a focus group technique with 33 participants. A qualitative research protocol that included informed consent, sociodemographic characterisation, observation guidelines, and a script with the dimensions explored was applied. Each group had an expert moderator and an observer. The audio of each session was recorded, transcribed, and analysed using ATLAS. Ti, version 8. Results: According to the handlers, the foods least consumed by the students are legumes, fresh vegetables, seafood, and some sauces. They estimate that food waste ranges from 25 to 100 kg per day. Women identify critical factors in the rejection of food by schoolchildren: aesthetics, smell, and taste of the dish; repetitiveness of the menu; and cultural aspects. As strategies to reduce waste, they propose varying the menu, incorporating foods familiar to schoolchildren, educating parents about the importance of offering healthy and sustainable food and preparations, and encouraging the participation of the entire educational community in the food process. Conclusions: The study revealed that food waste in school canteens is a complex problem that requires a multidimensional intervention that involves all actors in the school food system.

Ketogenesis promotes tolerance to Pseudomonas aeruginosa pulmonary infection.

Pseudomonas aeruginosa is a common cause of pulmonary infection. As a Gram-negative pathogen, it can initiate a brisk and highly destructive inflammatory response; however, most hosts become tolerant to the bacterial burden, developing chronic infection. Using a murine model of pneumonia, we demonstrate that this shift from inflammation to disease tolerance is promoted by ketogenesis. In response to pulmonary infection, ketone bodies are generated in the liver and circulate to the lungs where they impose selection for P. aeruginosa strains unable to display surface lipopolysaccharide (LPS). Such keto-adapted LPS strains fail to activate glycolysis and tissue-damaging cytokines and, instead, facilitate mitochondrial catabolism of fats and oxidative phosphorylation (OXPHOS), which maintains airway homeostasis. Within the lung, P. aeruginosa exploits the host immunometabolite itaconate to further stimulate ketogenesis. This environment enables host-P. aeruginosa coexistence, supporting both pathoadaptive changes in the bacteria and the maintenance of respiratory integrity via OXPHOS.

Lipidomics analysis reveals the effect of Sirex noctilio infestation on the lipid metabolism in Pinus radiata needles.

The Sirex noctilio's climatic adaption and rapid proliferation have caused Pinus mortality worldwide. The infestation combines the early effect of female S. noctilio gland secretion and the spreading symbiotic fungus Amylostereum areolatum. 'Lipidomics' is the study of all non-water-soluble components of the metabolome. Most of these non-water-soluble compounds correspond to lipids which can provide information about a biological activity, an organelle, an organism, or a disease. Using HPLC-MS/MS based lipidomics, 122 lipids were identified in P. radiata needles during S. noctilio infestation. Phosphatidic acids, N-acylethanolamines, and phosphatidylinositol-ceramides accumulated in infested trees could suggest a high level of phospholipases activities. The phosphatidylcholines were the most down-regulated species during infection, which could also suggest that they may be used as a substrate for up-regulated lipids. The accumulation of very long-chain fatty acids and long-chain fatty acids during the infestation could imply the tree defense response to create a barrier in the drilled zone to avoid larvae development and fungus proliferation. Also, the growth arrest phase of the trees during the prolonged infestation suggests a resistance response, regulated by the accumulation of NAE, which potentially shifts the tree energy to respond to the infestation.

Sostenibilidad alimentaria y prevalencia de consumo de preparaciones tradicionales y típicas en hogares del centro-sur de Chile / Food sustainability and prevalence of consumption of traditional and typical preparations in households in central-southern Chile

Introducción. Las preparaciones tradicionales típicas incluyen ingredientes mínimamente procesados, provenientes de la agricultura local, raíces y tradiciones de un territorio. A nivel mundial la población ha cambiado sus patrones dietéticos, incorporando alimentos ultraprocesados impactando la salud poblacional y planetaria. Objetivo. Identificar la sostenibilidad y la prevalencia de consumo de preparaciones tradicionales típicas chilenas en hogares de una región de la zona centro-sur de Chile. Materiales y Métodos. Diseño transversal descriptivo, con una muestra de 104 hogares mediante muestreo no probabilístico por conveniencia. Se identificó la sostenibilidad de 34 preparaciones tradicionales típicas chilenas, aplicando el sistema NOVA de clasificación de los alimentos según grado y tipo de procesamiento. Las preparaciones se clasificaron en sostenibles (>60% de ingredientes del grupo 1), medianamente sostenibles (50-60%) y no sostenibles (<50%). El estudio contó con la aprobación del Comité de Bioética de la Universidad del Bío-Bío. El análisis de datos consideró frecuencias, porcentajes e IC95% en STATA 17.0. Resultados. El 64% de las preparaciones fueron clasificadas como sostenibles, el 23% medianamente sostenibles y un 13% como no sostenibles. Las preparaciones sostenibles son las más consumidas por los hogares, especialmente aquellas que incluyen legumbres. Las preparaciones no sostenibles consumidas en los hogares se basan en harina refinada, manteca vegetal hidrogenado y/o aceites. Conclusiones. La sostenibilidad de las preparaciones tradicionales típicas chilenas depende de sus ingredientes. Las preparaciones sostenibles son las más consumidas por los hogares de esta región, y se basan en alimentos naturales o poco procesados(AU)

Introduction. Typical traditional preparations include minimally processed ingredients from local agriculture, roots and traditions of a territory. Globally, the population has changed its dietary patterns, incorporating ultra-processed foods, impacting population and planetary health. Objective. To identify the sustainability and prevalence of consumption of typical Chilean traditional preparations in households in central-southern Chile. Materials and Methods. Descriptive cross-sectional design, with a sample of 104 households using non-probabilistic convenience sampling. The sustainability of 34 typical Chilean traditional preparations was identif ied, applying the NOVA food classif ication system according to degree and type of processing. The preparations were classif ied as sustainable (>60% of group 1 ingredients), moderately sustainable (50- 60%) and non-sustainable (<50%). The Bioethics Committee of the Universidad del Bío-Bío approved the study. Data analysis considered f requencies, percentages and 95%CI in STATA 17.0. Results. 64% of the preparations were classif ied as sustainable, 23% moderately sustainable, and 13% non-sustainable. Sustainable preparations are the most consumed by households, especially those that include pulses. Unsustainable preparations households consume are based on ref ined flour, hydrogenated vegetable shortening and/or oils. Conclusions. The sustainability of typical Chilean traditional preparations depends on their ingredients. Sustainable preparations are the most consumed by households in this region and are based on natural or minimally processed foods(AU)

Staphylococcus aureus stimulates neutrophil itaconate production that suppresses the oxidative burst.

Neutrophils are critical in the host defense against Staphylococcus aureus, a major human pathogen. However, even in the setting of a robust neutrophil response, S. aureus can evade immune clearance. Here, we demonstrate that S. aureus impairs neutrophil function by triggering the production of the anti-inflammatory metabolite itaconate. The enzyme that synthesizes itaconate, Irg1, is selectively expressed in neutrophils during S. aureus pneumonia. Itaconate inhibits neutrophil glycolysis and oxidative burst, which impairs survival and bacterial killing. In a murine pneumonia model, neutrophil Irg1 expression protects the lung from excessive inflammation but compromises bacterial clearance. S. aureus is thus able to evade the innate immune response by targeting neutrophil metabolism and inducing the production of the anti-inflammatory metabolite itaconate.

The Impact of Highly Effective Modulator Therapy on Cystic Fibrosis Microbiology and Inflammation.

Highly effective cystic fibrosis (CF) transmembrane conductance regulator (CFTR) modulator therapy (HEMT) corrects the underlying molecular defect causing CF disease. HEMT decreases symptom burden and improves clinical metrics and quality of life for most people with CF (PwCF) and eligible cftr mutations. Improvements in measures of pulmonary health suggest that restoration of function of defective CFTR anion channels by HEMT not only enhances airway mucociliary clearance, but also reduces chronic pulmonary infection and inflammation. This article reviews the evidence for how HEMT influences the dynamic and interdependent processes of infection and inflammation in the CF airway, and what questions remain unanswered.

Stochastic tsunami modeling induced by kinematic complex sources.

In this study, local tsunami hazard due to seismic sources is evaluated in a stochastic framework. Several assumptions such as static passive generation, constant rake angle and source centroid (among others) are relaxed. Spatial uncertainties are modeled in a large set of scenarios. The proposed methodology is easy to implement and can be combined with other types of sources or hazards. Application in the Kuril-Kamchatkah trench shows a straightforward use of our methodology, producing simple hazard maps, which can be replicated in any region of the world.

Homology-based reconstruction of regulatory networks for bacterial and archaeal genomes.

Gene regulation is a key process for all microorganisms, as it allows them to adapt to different environmental stimuli. However, despite the relevance of gene expression control, for only a handful of organisms is there related information about genome regulation. In this work, we inferred the gene regulatory networks (GRNs) of bacterial and archaeal genomes by comparisons with six organisms with well-known regulatory interactions. The references we used are: Escherichia coli K-12 MG1655, Bacillus subtilis 168, Mycobacterium tuberculosis, Pseudomonas aeruginosa PAO1, Salmonella enterica subsp. enterica serovar typhimurium LT2, and Staphylococcus aureus N315. To this end, the inferences were achieved in two steps. First, the six model organisms were contrasted in an all-vs-all comparison of known interactions based on Transcription Factor (TF)-Target Gene (TG) orthology relationships and Transcription Unit (TU) assignments. In the second step, we used a guilt-by-association approach to infer the GRNs for 12,230 bacterial and 649 archaeal genomes based on TF-TG orthology relationships of the six bacterial models determined in the first step. Finally, we discuss examples to show the most relevant results obtained from these inferences. A web server with all the predicted GRNs is available at https://regulatorynetworks.unam.mx/ or http://132.247.46.6/.

Sirex noctilio infestation led to inevitable pine death despite activating pathways involved in tolerance.

Defense-related metabolome traits in pine species after infestation by Sirex noctilio are largely unknown, despite, in most cases, trees being overwhelmed. Using LC-MS-based untargeted metabolomics, we revealed the systemic metabolic changes induced by this insect in 14-year-old Pinus radiata trees, the most affected species worldwide. An immediate metabolome alteration was expressed in needles after infestation, including the up-regulation of flavonols, flavan-3-ols, oxyneolignans, auxins, proline, and tryptophan, among others. The flavan-3-ols (catechin and procyanidin B1) suggested a rapidly induced photoprotection mechanism aided by diverting proline as an alternative substrate for respiration to compensate for the progressive chlorosis that degrades photosystems. Meanwhile, glutathione, glutamate, and ascorbate levels significantly dropped in needles, which may indicate the critical oxidative stress that trees had to face since the onset of the infestation. They were not fully replenished after long-term infestation, and redox homeostasis was probably not achieved, compromising tree survival. Nevertheless, a huge auxins overexpression detected in needles throughout the infestation may reflect tolerance against the premature senescence caused by the woodwasp venom. In contrast, the metabolome of wood tissues remained initially unchanged, although it seems to collapse after three months. Overall, the metabolomics strategy adopted in this work evidenced its usefulness in uncovering the fundamental roles of plants' chemical defense that govern interactions with specific stressors.

Immunometabolic crosstalk during bacterial infection.

Following detection of bacteria, macrophages switch their metabolism from oxidative respiration through the tricarboxylic acid cycle to high-rate aerobic glycolysis. This immunometabolic shift enables pro-inflammatory and antimicrobial responses and is facilitated by the accumulation of fatty acids, tricarboxylic acid-derived metabolites and catabolism of amino acids. Recent studies have shown that these immunometabolites are co-opted by pathogens as environmental cues for expression of virulence genes. We review mechanisms by which host immunometabolites regulate bacterial pathogenicity and discuss opportunities for the development of therapeutics targeting metabolic host-pathogen crosstalk.

Pseudomonas aeruginosa Consumption of Airway Metabolites Promotes Lung Infection.

Prevailing dogma indicates that the lung of cystic fibrosis (CF) individuals is infected by multiple pathogens due to the abundant accumulation of mucus, which traps most of inhaled organisms. However, this hypothesis does not explain how specific opportunists, like Pseudomonas aeruginosa, are selected in the CF lung to cause chronic disease. This strongly suggests that other factors than mucus are accrued in the human airway and might predispose to bacterial disease, especially by P. aeruginosa. In this review we discuss the role of macrophage metabolites, like succinate and itaconate, in P. aeruginosa pneumonia. We analyze how dysfunction of the CF transmembrane conductance regulator (CFTR) favors release of these metabolites into the infected airway, and how P. aeruginosa exploits these elements to induce transcriptomic and metabolic changes that increase its capacity to cause intractable disease. We describe the host and pathogen pathways associated with succinate and itaconate catabolism, mechanisms of bacterial adaptation to these determinants, and suggest how both experimental settings and future therapies should consider macrophage metabolites abundance to better study P. aeruginosa pathogenesis.

Large slip, long duration, and moderate shaking of the Nicaragua 1992 tsunami earthquake caused by low near-trench rock rigidity.

Large earthquake ruptures propagating up to areas close to subduction trenches are infrequent, but when they occur, they heavily displace the ocean seafloor originating destructive tsunamis. The current paradigm is that the large seafloor deformation is caused by local factors reducing friction and increasing megathrust fault slip, or prompting the activation of ancillary faults or energy sources. As alternative to site-specific models, it has been proposed that large shallow slip could result from depth-dependent rock rigidity variations. To confront both hypotheses, here, we map elastic rock properties across the rupture zone of the M S7.0-M W7.7 1992 Nicaragua tsunami earthquake to estimate a property-compatible finite fault solution. The obtained self-consistent model accounts for trenchward increasing slip, constrains stress drop, and explains key tsunami earthquake characteristics such as long duration, high-frequency depletion, and magnitude discrepancy. The confirmation that these characteristics are all intrinsic attributes of shallow rupture opens new possibilities to improve tsunami hazard assessment.

Automated generation of context-specific gene regulatory networks with a weighted approach in Drosophila melanogaster.

The regulation of gene expression is a key factor in the development and maintenance of life in all organisms. Even so, little is known at whole genome scale for most genes and contexts. We propose a method, Tool for Weighted Epigenomic Networks in Drosophila melanogaster (Fly T-WEoN), to generate context-specific gene regulatory networks starting from a reference network that contains all known gene regulations in the fly. Unlikely regulations are removed by applying a series of knowledge-based filters. Each of these filters is implemented as an independent module that considers a type of experimental evidence, including DNA methylation, chromatin accessibility, histone modifications and gene expression. Fly T-WEoN is based on heuristic rules that reflect current knowledge on gene regulation in D. melanogaster obtained from the literature. Experimental data files can be generated with several standard procedures and used solely when and if available. Fly T-WEoN is available as a Cytoscape application that permits integration with other tools and facilitates downstream network analysis. In this work, we first demonstrate the reliability of our method to then provide a relevant application case of our tool: early development of D. melanogaster. Fly T-WEoN together with its step-by-step guide is available at https://weon.readthedocs.io.

Host-bacteria metabolic crosstalk drives S. aureus biofilm.

Staphylococcus aureus is a prominent pathogen that can cause intractable lung infections in humans. S. aureus persists in the airway despite inflammation and immune cell recruitment by adapting to host-derived antimicrobial factors. A key component of the immune response to infection are host metabolites that regulate inflammation and bacterial survival. In our recent paper (Tomlinson et al., Nat Commun, doi: 10.1038/s41467-021-21718-y), we demonstrated that S. aureus induces the production of the immunoregulatory metabolite itaconate in airway immune cells by stimulating mitochondrial oxidant stress. Itaconate in turn inhibited S. aureus glycolysis and growth, and promoted carbon flux through bacterial metabolic pathways that support biofilm production. These itaconate-induced metabolic changes were recapitulated in a longitudinal series of clinical isolates from a patient with chronic staphylococcal lung infections, demonstrating a role for host immunometabolism in driving bacterial persistence during long-term staphylococcal lung infections.

Pilot-plant scale extraction of phenolic compounds from grape canes: Comprehensive characterization by LC-ESI-LTQ-Orbitrap-MS.

Grape canes, also named vine shoots, are well-known viticultural byproducts containing high levels of phenolic compounds, which are associated with a broad range of health benefits. In this work, grape canes (Vitis vinifera cv. Pinot noir) were extracted in a 750 L pilot-plant reactor under the following conditions: temperature 80 °C, time 100 min, solid/liquid ratio 1:10. The comprehensive characterization of grape cane phenolic compounds was performed by liquid chromatography coupled to high-resolution/accurate mass measurement LTQ-Orbitrap mass spectrometry. A total of 44 compounds were identified and, 26 of them also quantified, consisting of phenolic acids and aldehydes (17), flavonoids (12), and stilbenoids (15). The most abundant class of phenolics were stilbenoids, among which (E)-ε-viniferin predominated. The phenolic profile of grape canes obtained using pilot plant extraction differed significantly from the results of laboratory-scale studies obtained previously. Additionally, we observed a high antioxidant capacity of grape cane pilot-plant extract measured by the radical antioxidant scavenging potential (ABTS+) (2209 ± 125 µmol TE/g DW) and oxygen radical absorbance capacity using fluorescein (ORAC-FL) (4612 ± 155 µmol TE/g DW). Grape cane pilot-plant extract for their phenolic profile may be used as a by-product for the development of novel nutraceutical and pharmaceutical products, improving the value and the sustainability of these residues.

Staphylococcus aureus induces an itaconate-dominated immunometabolic response that drives biofilm formation.

Staphylococcus aureus is a prominent human pathogen that readily adapts to host immune defenses. Here, we show that, in contrast to Gram-negative pathogens, S. aureus induces a distinct airway immunometabolic response dominated by the release of the electrophilic metabolite, itaconate. The itaconate synthetic enzyme, IRG1, is activated by host mitochondrial stress, which is induced by staphylococcal glycolysis. Itaconate inhibits S. aureus glycolysis and selects for strains that re-direct carbon flux to fuel extracellular polysaccharide (EPS) synthesis and biofilm formation. Itaconate-adapted strains, as illustrated by S. aureus isolates from chronic airway infection, exhibit decreased glycolytic activity, high EPS production, and proficient biofilm formation even before itaconate stimulation. S. aureus thus adapts to the itaconate-dominated immunometabolic response by producing biofilms, which are associated with chronic infection of the human airway.

Airway immunometabolites fuel Pseudomonas aeruginosa infection.

Pulmonary infections are associated with a brisk inflammatory reaction to bacterial surface components. Lipopolysaccharides (LPS) trigger macrophage activation and release of mitochondrial metabolites that control the intensity of the immune response. Whereas succinate induces oxidative stress (ROS), HIF1α stabilization, glycolysis and IL-1ß release, itaconate suppresses inflammation by inhibiting succinate oxidation, glycolytic flux and promoting anti-oxidant Nrf2-HO-1 functions. P. aeruginosa is a major pathogen associated with acute and chronic lung infection. Although both secreted toxins, LPS and proteases are key factors to establish acute P. aeruginosa pneumonia, lack of these components in chronic P. aeruginosa isolates suggest these organisms exploit other mechanisms to adapt and persist in the lung. Upon inhalation, P. aeruginosa strains trigger airway macrophage reprograming and bacterial variants obtained from acutely and chronically infected subjects exhibit metabolic adaptation consistent with succinate and itaconate assimilation; namely, high expression of extracellular polysaccharides (EPS), reduced lptD-LPS function, increased glyoxylate shunt (GS) activity and substantial biofilm production. In this review we discuss recent findings illustrating how P. aeruginosa induces and adapts to macrophage metabolites in the human lung, and that catabolism of succinate and itaconate contribute to their formidable abilities to tolerate oxidative stress, phagocytosis and immune clearance.