Diversity, Complexity, and Specificity of Bacterial Lipopolysaccharide (LPS) Structures Impacting Their Detection and Quantification.

Endotoxins are toxic lipopolysaccharides (LPSs), extending from the outer membrane of Gram-negative bacteria and notorious for their toxicity and deleterious effects. The comparison of different LPSs, isolated from various Gram-negative bacteria, shows a global similar architecture corresponding to a glycolipid lipid A moiety, a core oligosaccharide, and outermost long O-chain polysaccharides with molecular weights from 2 to 20 kDa. LPSs display high diversity and specificity among genera and species, and each bacterium contains a unique set of LPS structures, constituting its protective external barrier. Some LPSs are not toxic due to their particular structures. Different, well-characterized, and highly purified LPSs were used in this work to determine endotoxin detection rules and identify their impact on the host. Endotoxin detection is a major task to ensure the safety of human health, especially in the pharma and food sectors. Here, we describe the impact of different LPS structures obtained under different bacterial growth conditions on selective LPS detection methods such as LAL, HEK-blue TLR-4, LC-MS2, and MALDI-MS. In these various assays, LPSs were shown to respond differently, mainly attributable to their lipid A structures, their fatty acid numbers and chain lengths, the presence of phosphate groups, and their possible substitutions.

Study of Intra-Chamber Processes in Solid Rocket Motors by Fiber Optic Sensors.

In this study, an experimental study of the burning rate of solid fuel in a model solid propellant rocket motor (SRM) E-5-0 was conducted using a non-invasive control method with fiber-optic sensors (FOSs). Three sensors based on the Mach-Zehnder interferometer (MZI), fixed on the SRM E-5-0, recorded the vibration signal during the entire cycle of solid fuel burning. The results showed that, when using MZI sensors, the non-invasive control of solid fuel burnout is made possible both by recording the time of arrival of the combustion front to the sensor and by analyzing the peaks on the spectrogram of the recorded FOS signal. The main mode of acoustic vibrations of the chamber of the model SRM is longitudinal, and it changes with time, depending on the chamber length. Longitudinal modes of the combustion chamber were detected by MZI only after the combustion front passed its fixing point, and the microphone was unable to register them at all. The results showed that the combustion rate was practically constant after the first second, which was confirmed by the graph of the pressure versus time at the nozzle exit.

Regulation of waaH by PhoB during Pi Starvation Promotes Biofilm Formation by Escherichia coli O157:H7.

In open environments such as water, enterohemorrhagic Escherichia coli O157:H7 responds to inorganic phosphate (Pi) starvation by inducing the Pho regulon controlled by PhoB. This activates the phosphate-specific transport (Pst) system that contains a high-affinity Pi transporter. In the Δpst mutant, PhoB is constitutively activated and regulates the expression of genes in the Pho regulon. Here, we show that Pi starvation and deletion of the pst system enhance E. coli O157:H7 biofilm formation. Among differentially expressed genes of EDL933 grown under Pi starvation conditions and in the Δpst mutant, we have found that a member of the PhoB regulon, waaH, predicted to encode a glycosyltransferase, was highly expressed. Interestingly, WaaH contributed to biofilm formation of E. coli O157:H7 during both Pi starvation and in the Δpst mutant. In the Δpst mutant, the presence of waaH was associated with lipopolysaccharide (LPS) R3 core type modifications, whereas in E. coli O157:H7, waaH overexpression had no effect on LPS structure during Pi starvation. Therefore, waaH participates in E. coli O157:H7 biofilm formation during Pi starvation, but its biochemical role remains to be clarified. This study highlights the importance of the Pi starvation stress response to biofilm formation, which may contribute to the persistence of E. coli O157:H7 in the environment.IMPORTANCE Enterohemorrhagic Escherichia coli O157:H7 is a human pathogen that causes bloody diarrhea that can result in renal failure. Outside of mammalian hosts, E. coli O157:H7 survives for extended periods of time in nutrient-poor environments, likely as part of biofilms. In E. coli K-12, the levels of free extracellular Pi affect biofilm formation; however, it was unknown whether Pi influences biofilm formation by E. coli O157:H7. Our results show that upon Pi starvation, PhoB activates waaH expression, which favors biofilm formation by E. coli O157:H7. These findings suggest that WaaH is a target for controlling biofilm formation. Altogether, our work demonstrates how adaptation to Pi starvation allows E. coli O157:H7 to occupy different ecological niches.

Structure and dynamics of high-temperature strontium aluminosilicate melts.

We report the study of high-temperature melts (1600-2300 °C) and related glasses in the SrO-Al2O3-SiO2 phase diagram considering three series: (i) depolymerized ([SrO]/[Al2O3] = 3); (ii) fully polymerized ([SrO]/[Al2O3] = 1); and (iii) per-aluminous ([SrO]/[Al2O3] < 1). By considering the results from high-temperature 27Al NMR and high-temperature neutron diffraction, we demonstrate that the structure of the polymerized melts is controlled by a close-to-random distribution of Al and Si in the tetrahedral sites, while the depolymerized melts show smaller rings with a possible loss of non-bridging oxygens on AlO4 units during cooling for high-silica compositions. A few five-fold coordinated VAl sites are present in all compositions, except per-aluminous ones where high amounts of high-coordinated aluminium are found in glasses and melts with complex temperature dependence. In high-temperature melts, strontium has a coordination number of 8 or less, i.e. less than in the corresponding glasses. The dynamics of high-temperature melts were studied from 27Al NMR relaxation and compared to macroscopic shear viscosity data. These methods provide correlation times in close agreement. At very high temperatures, the NMR correlation times can be related to the oxygen self-diffusion coefficient, and we show a decrease of the latter with increasing Si/(Al + Si) ratios for polymerized melts with no compositional dependence for depolymerized ones. The dominant parameter controlling the temperature dependence of the aluminum environment of all melts is the distribution of Al-(OSi)p(OAl)(4-p) units.

Quantum Dot Emission Driven by Mie Resonances in Silicon Nanostructures.

Resonant dielectric nanostructures represent a promising platform for light manipulation at the nanoscale. In this paper, we describe an active photonic system based on Ge(Si) quantum dots coupled to silicon nanodisks. We show that Mie resonances govern the enhancement of the photoluminescent signal from embedded quantum dots due to a good spatial overlap of the emitter position with the electric field of Mie modes. We identify the coupling mechanism, which allows for engineering the resonant Mie modes through the interaction of several nanodisks. In particular, the mode hybridization in a nanodisk trimer results in an up to 10-fold enhancement of the luminescent signal due to the excitation of resonant antisymmetric magnetic and electric dipole modes.

Structural and biological characteristics of different forms of V. filiformis lipid A: use of MS to highlight structural discrepancies.

Vitreoscilla filiformis is a Gram-negative bacterium isolated from spa waters and described for its beneficial effects on the skin. We characterized the detailed structure of its lipopolysaccharide (LPS) lipid A moiety, an active component of the bacterium that contributes to the observed skin activation properties. Two different batches differing in postculture cell recovery were tested. Chemical analyses and mass spectra, obtained before and after mild-alkali treatments, revealed that these lipids A share the common bisphosphorylated ß-(1→6)-linked d-glucosamine disaccharide with hydroxydecanoic acid in an amide linkage. Short-chain FAs, hydroxydecanoic and dodecanoic acid, were found in a 2:1 ratio. The two lipid A structures differed by the relative amount of the hexa-acyl molecular species and phosphoethanolamine substitution of the phosphate groups. The two V. filiformis LPS batches induced variable interleukin-6 and TNF-α secretion by stimulated myelomonocytic THP-1 cells, without any difference in reactive oxygen species production or activation of caspase 3/7. Other different well-known highly purified LPS samples were characterized structurally and used as standards. The structural data obtained in this work explain the low inflammatory response observed for V. filiformis LPS and the previously demonstrated beneficial effects on the skin.

Combination of acoustic levitation with small angle scattering techniques and synchrotron radiation circular dichroism. Application to the study of protein solutions.

BACKGROUND: The acoustic levitation technique is a useful sample handling method for small solid and liquids samples, suspended in air by means of an ultrasonic field. This method was previously used at synchrotron sources for studying pharmaceutical liquids and protein solutions using x-ray diffraction and small angle x-ray scattering (SAXS). METHODS: In this work we combined for the first time this containerless method with small angle neutron scattering (SANS) and synchrotron radiation circular dichroism (SRCD) to study the structural behavior of proteins in solutions during the water evaporation. SANS results are also compared with SAXS experiments. RESULTS: The aggregation behavior of 45µl droplets of lysozyme protein diluted in water was followed during the continuous increase of the sample concentration by evaporating the solvent. The evaporation kinetics was followed at different drying stage by SANS and SAXS with a good data quality. In a prospective work using SRCD, we also studied the evolution of the secondary structure of the myoglobin protein in water solution in the same evaporation conditions. CONCLUSIONS: Acoustic levitation was applied for the first time with SANS and the high performances of the used neutron instruments made it possible to monitor fast container-less reactions in situ. A preliminary work using SRCD shows the potentiality of its combination with acoustic levitation for studying the evolution of the protein structure with time. GENERAL SIGNIFICANCE: This multi-techniques approach could give novel insights into crystallization and self-assembly phenomena of biological compound with promising potential applications in pharmaceutical, food and cosmetics industry. This article is part of a Special Issue entitled "Science for Life" Guest Editor: Dr. Austen Angell, Dr. Salvatore Magazù and Dr. Federica Migliardo.

Bordetella holmesii: Lipid A Structures and Corresponding Genomic Sequences Comparison in Three Clinical Isolates and the Reference Strain ATCC 51541.

Bordetella holmesii can cause invasive infections but can also be isolated from the respiratory tract of patients with whooping-cough like symptoms. For the first time, we describe the lipid A structure of B. holmesii reference strain ATCC 51541 (alias NCTC12912 or CIP104394) and those of three French B. holmesii clinical isolates originating from blood (Bho1) or from respiratory samples (FR4020 and FR4101). They were investigated using chemical analyses, gas chromatography-mass spectrometry (GC-MS), and matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS). The analyses revealed a common bisphosphorylated ß-(1→6)-linked d-glucosamine disaccharide with hydroxytetradecanoic acid in amide linkages. Similar to B. avium, B. hinzii and B. trematum lipids A, the hydroxytetradecanoic acid at the C-2' position are carrying in secondary linkage a 2-hydroxytetradecanoic acid residue resulting of post-traductional biosynthesis modifications. The three clinical isolates displayed characteristic structural traits compared to the ATCC 51541 reference strain: the lipid A phosphate groups are more or less modified with glucosamine in the isolates and reference strain, but the presence of 10:0(3-OH) is only observed in the isolates. This trait was only described in B. pertussis and B. parapertussis strains, as well as in B. petrii isolates by the past. The genetic bases for most of the key structural elements of lipid A were analyzed and supported the structural data.

Minor modifications to the phosphate groups and the C3' acyl chain length of lipid A in two Bordetella pertussis strains, BP338 and 18-323, independently affect Toll-like receptor 4 protein activation.

Lipopolysaccharides (LPS) of Bordetella pertussis are important modulators of the immune system. Interaction of the lipid A region of LPS with the Toll-like receptor 4 (TLR4) complex causes dimerization of TLR4 and activation of downstream nuclear factor κB (NFκB), which can lead to inflammation. We have previously shown that two strains of B. pertussis, BP338 (a Tohama I-derivative) and 18-323, display two differences in lipid A structure. 1) BP338 can modify the 1- and 4'-phosphates by the addition of glucosamine (GlcN), whereas 18-323 cannot, and 2) the C3' acyl chain in BP338 is 14 carbons long, but only 10 or 12 carbons long in 18-323. In addition, BP338 lipid A can activate TLR4 to a greater extent than 18-323 lipid A. Here we set out to determine the genetic reasons for the differences in these lipid A structures and the contribution of each structural difference to the ability of lipid A to activate TLR4. We show that three genes of the lipid A GlcN modification (Lgm) locus, lgmA, lgmB, and lgmC (previously locus tags BP0399-BP0397), are required for GlcN modification and a single amino acid difference in LpxA is responsible for the difference in C3' acyl chain length. Furthermore, by introducing lipid A-modifying genes into 18-323 to generate isogenic strains with varying penta-acyl lipid A structures, we determined that both modifications increase TLR4 activation, although the GlcN modification plays a dominant role. These results shed light on how TLR4 may interact with penta-acyl lipid A species.

Controlled electroactive release from solid-state conductive elastomer electrodes.

This work highlights the development of a conductive elastomer (CE) based electrophoretic platform that enables the transfer of charged molecules from a solid-state CE electrode directly to targeted tissues. Using an elastomer-based electrode containing poly (3,4-ethylenedioxythiophene) nanowires, controlled electrophoretic delivery of methylene blue (MB) and fluorescein (FLSC) was achieved with applied voltage. Electroactive release of positively charged MB and negatively charged FLSC achieved 33.19 ± 6.47 µg release of MB and 22.36 ± 3.05 µg release of FLSC, a 24 and 20-fold increase in comparison to inhibitory voltages over 1 h. Additionally, selective, and sequential release of the two oppositely charged molecules from a single CE device was demonstrated, showing the potential of this device to be used in multi-drug treatments.

Tuning the Luminescence Response of an Air-Hole Photonic Crystal Slab Using Etching Depth Variation.

Detailed studies of the luminescent properties of the Si-based 2D photonic crystal (PhC) slabs with air holes of various depths are reported. Ge self-assembled quantum dots served as an internal light source. It was obtained that changing the air hole depth is a powerful tool which allows tuning of the optical properties of the PhC. It was shown that increasing the depth of the holes in the PhC has complex influences on its overall photoluminescence (PL) response due to the simultaneous influences of counteracting factors. As a result, the maximal increase in the PL signal of more than two orders of magnitude was obtained for some intermediate, but not full, depth of the PhC's air holes. It was demonstrated that it is possible to engineer the PhC band structure in such a way as to construct specific states, namely bound states in continuum (BIC), with specially designed dispersion curves being relatively flat. In this case, such states manifest themselves as sharp peaks in the PL spectra, and have high Q-factors which are larger than those of radiative modes and other BIC modes without such a flat dispersion characteristic.

A TLR4 Agonist Induces Osteosarcoma Regression by Inducing an Antitumor Immune Response and Reprogramming M2 Macrophages to M1 Macrophages.

Osteosarcoma (OsA) has limited treatment options and stagnant 5-year survival rates. Its immune microenvironment is characterized by a predominance of tumor-associated macrophages (TAMs), whose role in OsA progression remain unclear. Nevertheless, immunotherapies aiming to modulate macrophages activation and polarization could be of interest for OsA treatment. In this study, the antitumor effect of a liposome-encapsulated chemically detoxified lipopolysaccharide (Lipo-MP-LPS) was evaluated as a therapeutic approach for OsA. Lipo-MP-LPS is a toll-like receptor 4 (TLR4) agonist sufficiently safe and soluble to be IV administered at effective doses. Lipo-MP-LPS exhibited a significant antitumor response, with tumor regression in 50% of treated animals and delayed tumor progression in the remaining 50%. The agent inhibited tumor growth by 75%, surpassing the efficacy of other immunotherapies tested in OsA. Lipo-MP-LPS modulated OsA's immune microenvironment by favoring the transition of M2 macrophages to M1 phenotype, creating a proinflammatory milieu and facilitating T-cell recruitment and antitumor immune response. Overall, the study demonstrates the potent antitumor effect of Lipo-MP-LPS as monotherapy in an OsA immunocompetent model. Reprogramming macrophages and altering the immune microenvironment likely contribute to the observed tumor control. These findings support the concept of immunomodulatory approaches for the treatment of highly resistant tumors like OsA.

Emission Enhancement of Ge/Si Quantum Dots in Hybrid Structures with Subwavelength Lattice of Al Nanodisks.

The effects of resonance interaction of plasmonic and photonic modes in hybrid metal-dielectric structures with square Al nanodisk lattices coupled with a Si waveguide layer were investigated using micro-photoluminescence (micro-PL) spectroscopy. As radiation sources, GeSi quantum dots were embedded in the waveguide. A set of narrow PL peaks superimposed on the broad bands were observed in the range of quantum dot emissions. At optimal parameters of Al nanodisks lattices, almost one order increasing of PL intensity was obtained. The experimental PL spectra are in good agreement with results of theoretical calculations. The realization of high-quality bound states in the continuum was confirmed by a comparative analysis of the experimental spectra and theoretical dispersion dependences. The results demonstrated the perspectives of these type structures for a flat band realization and supporting the slow light.

A systemically administered detoxified TLR4 agonist displays potent antitumor activity and an acceptable tolerance profile in preclinical models.

Bacterial lipopolysaccharides (LPS) are potent innate immunostimulants targeting the Toll-like receptor 4 (TLR4), an attractive and validated target for immunostimulation in cancer therapy. Although LPS possess anti-tumor activity, toxicity issues prevent their systemic administration at effective doses in humans. We first demonstrated that LPS formulated in liposomes preserved a potent antitumor activity per se upon systemic administration in syngeneic models, and significantly enhance the antitumor activity of the anti-CD20 antibody rituximab in mice xenografted with the human RL lymphoma model. Liposomal encapsulation also allowed a 2-fold reduction in the induction of pro-inflammatory cytokines by LPS. Mice receiving an intravenous administration demonstrated a significant increase of neutrophils, monocytes and macrophages at the tumor site as well as an increase of macrophages in spleen. Further, we chemically detoxified LPS to obtain MP-LPS that was associated with a 200-fold decrease in the induction of proinflammatory cytokines. When encapsulated in a clinically approved liposomal formulation, toxicity, notably pyrogenicity (10-fold), was limited while the antitumor activity and immunoadjuvant effect were maintained. This improved tolerance profile of liposomal MP-LPS was associated with the preferential activation of the TLR4-TRIF pathway. Finally, in vitro studies demonstrated that stimulation with encapsulated MP-LPS reversed the polarization of M2 macrophages towards an M1 phenotype, and a phase 1 trial in healthy dogs validated its tolerance upon systemic administration up to very high doses (10µg/kg). Altogether, our results demonstrate the strong therapeutic potential of MPLPS formulated in liposomes as a systemically active anticancer agent, supporting its evaluation in patients with cancer.

Changes in chemiluminescence of whole blood of COPD patients treated with Hypoxen and effects of C60 fullerenes on blood chemiluminescence.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an inflammatory disease associated with reactive oxygen species (ROS) production. The aim of this study was to evaluate the effect of Hypoxen treatment and the effect of HyFnC60 on ROS production in patients' blood. MATERIAL/METHODS: ROS production in blood was estimated using chemiluminescence (CL) measurement with CL-amplifiers: luminol (LM), LM + zymosan (ZM) or lucigenin (LC) in the presence or absence of hydrated fullerenes (HyFnC60) added to blood in low concentrations. RESULTS: In all the patients with COPD in remission phase with Hypoxen prescription, the LM-dependent CL (LM-CL) with ZM and LC-enhanced CL (LC-CL) decreased after the treatment. Parameters of CL and effects of HyFnC60 upon them depended on blood state. Addition of HyFnC60 to blood decreased data scattering and helped to improve discrimination between different groups of patients. Using the discriminator analysis, we found the most important time-points in the kinetic curves of CL for classification of patients into groups (eg, COPD patients before and after treatment with Hypoxen; patients' blood with different sensitivity to HyFnC60 concentration). CONCLUSIONS: Monitoring of CL of non-diluted whole blood in COPD patients can be used for the estimation of the Hypoxen efficiency in complex therapy. Addition of HyFnC60 to blood increases sensitivity of the method.

Cognitive Abilities and Academic Achievement as Intercultural Competence Predictors in Russian School Students.

The development of intercultural competence (ICC) is important for the modern personality in an unstable and diverse world, but there is a lack of research on this phenomenon in the context of age, gender and intellectual differences. The purpose of the present exploratory study is to identify relations between ICC, cognitive abilities and academic achievements among Russian school students. The sample included 106 (55% female) students in the 9th grade of Moscow secondary school. ICC was measured with the author's modification of The Intercultural Sensitivity Scale by Khuhlaev and Chibisova, developed on the basis of the Developmental Model of Intercultural Sensitivity by Bennett. Cognitive abilities were determined with the School Test of Intellectual Development by Akimova et al. Academic achievements were evaluated using GPA. The findings of our research show that: (1) higher academic achievements and cognitive abilities usually characterize schoolchildren, who are not inclined to absolutize cultural differences and do not consider them to be barriers to intercultural interaction; (2) the most significant predictors of ICC features from the studied cognitive abilities are analogy and generalization, but generalization has opposite impacts in male and female students. This fact should be taken into account in the context of ICC developments, especially in male school students prone to ethnocentrism.

Interaction of Ge(Si) Self-Assembled Nanoislands with Different Modes of Two-Dimensional Photonic Crystal.

The interaction of Ge(Si)/SOI self-assembled nanoislands with modes of photonic crystal slabs (PCS) with a hexagonal lattice is studied in detail. Appropriate selection of the PCS parameters and conditions for collecting the photoluminescence (PL) signal allowed to distinguish the PCS modes of different physical nature, particularly the radiative modes and modes associated to the bound states in the continuum (BIC). It is shown that the radiative modes with relatively low Q-factors could provide a increase greater than an order of magnitude in the integrated PL intensity in the wavelength range of 1.3-1.55 µm compared to the area outside of PCS at room temperature. At the same time, the interaction of Ge(Si) islands emission with the BIC-related modes provides the peak PL intensity increase of more than two orders of magnitude. The experimentally measured Q-factor of the PL line associated with the symmetry-protected BIC mode reaches the value of 2600.

Enucleation vs. vaporization of benign prostatic hyperplasia: a head-to-head comparison of the various outcomes and complications. A systematic review and meta-analysis.

INTRODUCTION: Vaporization of the prostate (VP) and endoscopic enucleation of the prostate (EEP) are reliable and frequently used methods for BPO relief. Both surgeries utilize lasers and EAU recommends them in similar patient cohorts. Our objective was to compare intra- and perioperative results of patients who had undergone VP and EEP. EVIDENCE ACQUISITION: A systematic literature search was performed in three databases (MEDLINE, Web of Science and Scopus). The detailed search strategy is available at Prospero, CRD42020204739. Primary outcomes were functional results (IPSS, QoL, PVR, Qmax), and secondary outcomes were intraoperative results, postoperative PSA and prostate volume, complications, and recurrence rate. EVIDENCE SYNTHESIS: VP required less operative time compared to EEP, mean difference -5.51 (95% CI -7.52; -3.50). IPSS and Qmax for VP were worse after 12-month follow-up, mean difference 0.89 (95% CI 0.52; 1.27) and -3.7 (95% CI -4.56; -2.85), respectively, while QoL did not differ significantly. Postoperative PSA level was higher in the VP group, mean difference 2.28 (95% CI 2.00; 2.55). VP was associated with reduced Clavien-Dindo grade I (OR=4.16; 95% CI 2.96; 5.84) and grade II (OR=3.79; 95% CI 2.25; 6.39) complication rate, especially in terms of the percentage of blood transfusion and transient urinary incontinence. The rate of complications grade IIIa and higher was similar (3-6%). Reoperation rate was only estimated in one study and was significantly higher in the PVP group at 60 months of follow-up, 2.7% vs. 0%, P<0.05. CONCLUSIONS: EEP and VP share the efficacy and safety in BPH management. Our meta-analysis shows comparable complication rate in Clavien-Dindo III, VP superiority in operation time, and EEP superiority in long-term functional outcomes and PSA reduction.

A secondary metabolite acting as a signalling molecule controls Pseudomonas entomophila virulence.

Pseudomonas entomophila is an entomopathogenic bacterium that is lethal to Drosophila melanogaster within 1-2 days of ingestion of high doses. Flies orally infected with P. entomophila rapidly succumb despite the induction of both local and systemic immune responses. Recent studies suggest that its virulence relies on its ability to cause irreversible damages to the intestinal epithelium, in contrast to what is observed with milder pathogenic bacteria such as Erwinia carotovora carotovora Ecc15 or Pseudomonas aeruginosa PA14. The GacS/GacA two-component system plays a key role in P. entomophila pathogenicity. Here, we report the identification of the pvf genes, whose products are involved in production of a secondary metabolite involved in P. entomophila virulence. A pvf mutant is impaired in its ability to persist within the gut, to trigger the fly immune responses and to inflict gut damages. The expression of several genes is affected in a pvf mutant, independently of the Gac system. Moreover, growing a pvf mutant in medium supplemented with supernatant extracts from either the wild-type strain or a gacA mutant restore its pathogenicity. Collectively, our results indicate that we identified genes involved in the synthesis of a signalling molecule that controls P. entomophila virulence independently from the Gac system.

Structural modifications occurring in lipid A of Bordetella bronchiseptica clinical isolates as demonstrated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Bordetella bronchiseptica is a respiratory pathogen in mammal species and its cell surface lipopolysaccharide-endotoxin is a potent virulence factor. In order to better characterize the endotoxin structure to virulence relationships, we studied the lipid A structures of B. bronchiseptica isolates from human and rabbit origins as a function of their virulence phases. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has been widely used for the structural characterization of bacterial endotoxins and their lipid A moieties. This method combined with chemical analytical methods proved to be essential for the characterization of small samples and discrete but essential structural modifications. The occurrence of palmitate (C(16)) in the B. bronchiseptica lipid A structures is shown for the first time at two sites. Their presence was also demonstrated for the first time in correlation with the virulence phase of B. bronchiseptica clinical isolates. The recently identified glucosamine modifications of Bordetella lipids A are also reported in these isolates.