Desulfovibrio desulfuricans isolates from the gut of a single individual: Structural and biological lipid A characterization

AbstractThe levels of sulfate-reducing bacteria (SRB), including Desulfovibrionaceae, in the gut increase following a fat-enriched diet. Endotoxins from gut microbiota contribute to the inflammation process, leading to metabolic diseases. Thus, we sought to characterize the lipid A structures of Desulfovibrionaceae lipopolysaccharides (LPS) that are associated with the microbiota inflammatory properties. LPS variants were obtained from two SRB isolates from the gut of a single individual. These LPS variants shared similar lipid A moieties with Enterobacterial LPS, but differed from one another with regard to fatty-acid numbers and endotoxic activity. This first complete structural characterization of Desulfovibrio lipid A gives new insights into previously published data on Desulfovibrio lipid A biosynthesis. LPS microdiversity within SRBs illustrates how adaptation can influence pro-inflammatory potential

Processamento auditivo em crianças e adolescentes em situação de risco e vulnerabilidade

CONTEXT AND OBJECTIVE: Children and adolescents who live in situations of social vulnerability present a series of health problems. Nonetheless, affirmations that sensory and cognitive abnormalities are present are a matter of controversy. The aim of this study was to investigate aspects to auditory processing, through applying the brainstem auditory evoked potential (BAEP) and behavioral auditory processing tests to children living on the streets, and comparison with a control group. DESIGN AND SETTING: Cross-sectional study in the Laboratory of Auditory Processing, School of Medicine, Universidade de São Paulo. METHODS: The auditory processing tests were applied to a group of 27 individuals, subdivided into 11 children (7 to 10 years old) and 16 adolescents (11 to 16 years old), of both sexes, in situations of social vulnerability, compared with an age-matched control group of 10 children and 11 adolescents without complaints. The BAEP test was also applied to investigate the integrity of the auditory pathway. RESULTS: For both children and adolescents, there were significant differences between the study and control groups in most of the tests applied, with significantly worse performance in the study group, except in the pediatric speech intelligibility test. Only one child had an abnormal result in the BAEP test. CONCLUSIONS: The results showed that the study group (children and adolescents) presented poor performance in the behavioral auditory processing tests, despite their unaltered auditory brainstem pathways, as shown by their normal results in the BAEP test

The Drosophila immune system detects bacteria through specific peptidoglycan recognition

The Drosophila immune system discriminates between different classes of infectious microbes and responds with pathogen-specific defense reactions through selective activation of the Toll and the immune deficiency (Imd) signaling pathways. The Toll pathway mediates most defenses against Gram-positive bacteria and fungi, whereas the Imd pathway is required to resist infection by Gram-negative bacteria. The bacterial components recognized by these pathways remain to be defined. Here we report that Gram-negative diaminopimelic acid-type peptidoglycan is the most potent inducer of the Imd pathway and that the Toll pathway is predominantly activated by Gram-positive lysine-type peptidoglycan. Thus, the ability of Drosophila to discriminate between Gram-positive and Gram-negative bacteria relies on the recognition of specific forms of peptidoglycan

Micromethods for lipid a isolation and structural characterization.

International audienceLipopolysaccharides (LPSs) are major components of the external membrane of Gram-negative bacteria, and act as an effective permeability barrier. They are essentially composed of a hydrophilic polysaccharide region linked to an hydrophobic one, termed lipid A. Depending on their individual variable fine structures, they may be potent immunomodulators. Because of the structural importance and role of lipid A in bacterial pathogenesis, herein we describe two rapid practical micromethods for structural analysis. The first method allows the direct isolation of lipid A from whole bacteria cell mass; the second describes conditions for the sequential release of fatty acids, enabling the determination of their substitution position in the lipid A structure to be determined by matrix-assisted laser desorption/ionization mass spectrometry. Examples are given with reference to two major pathogens: Bordetella pertussis and Pseudomonas aeruginosa

Lipopolysaccharides: structure, function and bacterial identifications

Lipopolysaccharides (LPS) are the main components of the outer membrane of Gram-negative bacteria. They are glycolipids containing a lipid moiety called lipid A, more often made of a bis-phosphorylated glucosamine disaccharide, carrying fatty acids in ester and amide linkages. Lipid A is linked to a core oligosaccharide of about 10 sugars, substituted in the wild-type strains, by long-chain oligosaccharide repetitive units, extending outside the bacteria and representing their main antigens. In addition to determine the serotype of the bacterium, LPS are highly potent biological molecules, capable of eliciting at the level of minute amounts, beneficial, as well as deleterious activities

Etude de l'effet de la salinité sur la survie et le pouvoir infectieux de Rhizobia Nodulant les Acacias : Analyse de la structure des lipopolysaccharides

LE KREMLIN-B.- PARIS 11-BU Méd (940432101) / SudocSudocFranceF

Structure of the Bordetella trematum LPS O-chain subuit

NRC publication: Ye

A comparative study of the complete lipopolysaccharide structures and biosynthesis loci of Bordetella avium, B. hinzii, and B. trematum

International audienceA dozen species of human and animal pathogens have been described to date in the Bordetella genus, with the majority being respiratory tract pathogens. Bordetella avium lipopolysaccharides have been shown to be important virulence factors for this bird pathogen. B. hinzii is closely related to the B. avium species, but has also been isolated from humans. B. trematum is associated to ear and blood infections in humans. Its lipid A structure, the biological active moiety of LPS, was found to be closely related to those of B. avium and B. hinzii. It is important to unveil the subtle structural modifications orchestrated during the LPS biosynthetic pathway to better understand host adaptation. The present data are also important in the context of deciphering the virulence pathways of this important genus containing the major pathogens B. pertussis and B. parapertussis, responsible for whooping cough. We recently reported the isolated lipid A structures of the three presented species, following the previously identified O-chain structures. In the present study, we provide details on the free and O-chain-linked core oligosaccharides which were required to characterize the complete LPS structures. Data are presented here in relation to relevant biosynthesis genes. The present characterization of the three species is well illustrated by Matrix Assisted Laser Desorption Mass Spectrometry experiments, and data were obtained mainly on native LPS molecules for the first time. (C) 2018 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved

Micromethods for Isolation and Structural Characterization of Lipid A, and Polysaccharide Regions of Bacterial Lipopolysaccharides

Lipopolysaccharides (LPS) are major components of the external membrane of most Gram-negative bacteria, providing them with an effective permeability barrier. They are essentially composed of a hydrophilic polysaccharide region (PS) linked to a hydrophobic one, termed lipid A. The LPS polysaccharide moiety is divided into the core oligosaccharide (OS) and O-chain repetitive elements. Depending on their individual variable fine structures, LPS may be potent immunomodulators. The lipid A structure is a key determinant for LPS activity. However, the presence of the core region, or at least of the highly charged 3-deoxy-d-manno-oct-2-ulosonic acid molecules, is also important for preserving the native lipid A conformation within individual LPS molecules. We describe herein four rapid and practical micromethods for LPS, lipid A, and core OS structural analyses. The first method allows the direct isolation of lipid A from whole bacteria cell mass; the second describes conditions for the sequential release of fatty acids enabling the characterization of their substitution position in the lipid A backbone, to be determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The third one is a microscale procedure for the mass spectra screening of LPS, lipid A, and PS using triethylamine and citric acid. The fourth method is a chromatography procedure for Rough-type LPS on thin-layer-chromatography. These methods were developed to be coupled to mass-spectrometry (e.g., MALDI-MS) but can also be used with other analytical techniques (e.g., chromatography). Examples are given with reference to two major human pathogens: Bordetella pertussis and Pseudomonas aeruginosa; to one porcine pathogen: Actinobacillus pleuropneumoniae; and to commercial samples of Salmonella Minnesota Re595 LPS

Analyse structurale d'endotoxines bactériennes par spectrométrie de masse

Du point de vue chimique les endotoxines sont des lipopolysaccharides (LPS) constitués d une chaîne de sucres, caractéristique de chaque espèce bactérienne, et d une structure lipidique, le lipide A. Nous avons mis au point des méthodes d analyse structurale et de purification permettant des avancées importantes dans le domaine de la relation structure-activité. La première est une micro-méthode d analyse qui peut être appliquée à des quantités de bactéries de l ordre du milligramme. Elle conduit à l obtention de spectres de lipide A en une journée, au lieu d une semaine par les techniques classiques. La seconde permet d établir la structure et le positionnement des acides gras sur des quantités de lipide A minimes. Cette technologie a été appliquée aux lipides A de Citrobacter, une Entérobactérie responsable de maladies nosocomiales. Enfin, une méthode de purification permet d obtenir des échantillons hautement purifiés pour les tests biologiques. Elle permettra des comparaisons plus fiables entre différentes préparations purifiées dans différents laboratoires. Nous avons aussi mis en évidence la présence de nouveaux substituants au niveau de la région lipidique des lipides A du genre Bordetella, dont B. pertussis, la coqueluche. Il s agit de la présence d une glucosamine sur chaque phosphate dont l implication sur les activités biologiques seront majeurs au niveau de l action des peptides antibactériennes, car ces structures viennent neutraliser les charges du lipide A, la molécule responsable des activités endotoxiques du LPS.Endotoxins are lipopolysaccharides (LPS) made up of a lipid - (called lipid A) and a sugar - chain, both characteristic of each bacterial species. We developed methods of structure analysis and a purification method representing an important improvement for studies of structure/activity relationships. The first one is a micro analytical method that can be applied to milligram quantities of bacteria. With it, one can obtain spectra of the lipid A in one day, instead of a week as for previous techniques. The second one leads to the determination of the structure and the positions of the fatty acids with a little amount of lipid A. This technology was applied to Citrobacter, an Enterobacterium causing nosocomial diseases. A LPS purification method gave highly purified samples suitable for biological tests. This method will give preparation having identical activities in different laboratories. We also have established the existence of new constituents on the lipid A of B. bronchiseptica, from which the whooping cough originated. The presence of a glucosamine on the phosphate group(s) was demonstrated and its effect on the biological activities will be of high impact particularly on the activity of anti-bacterial peptides, as glucosamines neutralise the lipid A structure responsible for the endotoxic activities of LPS.ORSAY-PARIS 11-BU Sciences (914712101) / SudocSudocFranceF