Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica

The inhalation of airborne pollutants, such as asbestos or silica, is linked to inflammation of the lung, fibrosis, and lung cancer. How the presence of pathogenic dust is recognized and how chronic inflammatory diseases are triggered are poorly understood. Here, we show that asbestos and silica are sensed by the Nalp3 inflammasome, whose subsequent activation leads to interleukin-1beta secretion. Inflammasome activation is triggered by reactive oxygen species, which are generated by a NADPH oxidase upon particle phagocytosis. (NADPH is the reduced form of nicotinamide adenine dinucleotide phosphate.) In a model of asbestos inhalation, Nalp3-/- mice showed diminished recruitment of inflammatory cells to the lungs, paralleled by lower cytokine production. Our findings implicate the Nalp3 inflammasome in particulate matter-related pulmonary diseases and support its role as a major proinflammatory "danger" receptor

Web-based Implementation of Winter Maintenance Decision Support System Using GIS and Remote Sensing, May 2005

Winter maintenance, particularly snow removal and the stress of snow removal materials on public structures, is an enormous budgetary burden on municipalities and nongovernmental maintenance organizations in cold climates. Lately, geospatial technologies such as remote sensing, geographic information systems (GIS), and decision support tools are roviding a valuable tool for planning snow removal operations. A few researchers recently used geospatial technologies to develop winter maintenance tools. However, most of these winter maintenance tools, while having the potential to address some of these information needs, are not typically placed in the hands of planners and other interested stakeholders. Most tools are not constructed with a nontechnical user in mind and lack an easyto-use, easily understood interface. A major goal of this project was to implement a web-based Winter Maintenance Decision Support System (WMDSS) that enhances the capacity of stakeholders (city/county planners, resource managers, transportation personnel, citizens, and policy makers) to evaluate different procedures for managing snow removal assets optimally. This was accomplished by integrating geospatial analytical techniques (GIS and remote sensing), the existing snow removal asset management system, and webbased spatial decision support systems. The web-based system was implemented using the ESRI ArcIMS ActiveX Connector and related web technologies, such as Active Server Pages, JavaScript, HTML, and XML. The expert knowledge on snow removal procedures is gathered and integrated into the system in the form of encoded business rules using Visual Rule Studio. The system developed not only manages the resources but also provides expert advice to assist complex decision making, such as routing, optimal resource allocation, and monitoring live weather information. This system was developed in collaboration with Black Hawk County, IA, the city of Columbia, MO, and the Iowa Department of transportation. This product was also demonstrated for these agencies to improve the usability and applicability of the system.

Membrane and walls: who is master, who is servant?

Specialised plant cell types often locally modify their cell walls as part of a developmental program, as do cells that are challenged by particular environmental conditions. Modifications can include deposition of secondary cellulose, callose, cutin, suberin or lignin. Although the biosyntheses of cell wall components are more and more understood, little is known about the mechanisms that control localised deposition of wall materials. During metaxylem vessel differentiation, site-specific cell wall deposition is locally prevented by the microtubule depolymerising protein MIDD1, which disassembles the cytoskeleton and precludes the cellulose synthase complex from depositing cellulose. As a result, metaxylem vessel secondary cell wall appears pitted. How MIDD1 is tethered at the plasma membrane and how other cell wall polymers are locally deposited remain elusive. Casparian strips in the root endodermis represent a further example of local cell wall deposition. The recent discovery of the Casparian Strip membrane domain Proteins (CASPs), which are located at the plasma membrane and are important for the site-specific deposition of lignin during Casparian strip development, establishes the root endodermis as an attractive model system to study the mechanisms of localised cell wall modifications. How secondary modifications are modulated and monitored during development or in response to environmental changes is another question that still misses a complete picture.

Sensing, uptake and response to c4-dicarboxylic acids in pseudomonas aeruginosa pao1

C4-dicarboxylates are one of the preferred carbon and energy sources for the growth of P. aeruginosa, a ubiquitous and metabolically versatile bacterium. However, despite their importance, C4-dicarboxylates sensing and uptake systems were poorly understood in P. aeruginosa and only little information was available in the literature. In our work, the C4-dicarboxylate transport (Dct) system in P. aeruginosa was found to be composed of a novel two-component system, called DctB/DctD, regulating together with the sigma factor RpoN the expression of two newly identified C4-dicarboxylate transporters: DctA and DctPQM. Inactivation of the dct A, dctB or dctD gene caused a growth defect of the strain in minimal media supplemented with succinate, fumarate or malate, indicating their major role in Dct. However, residual growth of the dctA mutant in these media suggested the presence of redundant C4-dicarboxylate transporter(s). Tn5 insertion mutagenesis of the kdctA mutant, combined with a screening for growth on succinate, led to the identification of a second Dct system, the DctPQM transporter, belonging to the tripartite ATP-independent periplasmic (TRAP) family of carriers. AdctAAdctPQM double mutant showed no growth on malate and fumarate albeit residual growth on succinate suggested that additional transporters for succinate are present. Competition experiments demonstrated that the DctPQM carrier was more efficient than the DctA carrier for the utilization of succinate at μΜ concentrations, whereas DctA was the major transporter at mM concentrations. For the first time, high- and low-affinity uptake systems for succinate (DctA and DctPQM) are reported to function co-ordinately to transport C4- dicarboxylates. Most probably, the presence of redundant uptake systems contributes to the versatility of this bacterium. Next, the regulation of the Dct system was investigated. While performing a parallel study about the carbon catabolite repression (CCR) phenomenon in P. aeruginosa, a link between the CCR cascade (CbrAB/CrcZ/Crc) and the Dct system was observed. Crc is a translational repressor acting when preferred carbon sources (like C4-dicarboxylates) are present. CrcZ is a small RNA acting as a functional antagonist of Crc and induced by the CbrA/CbrB two-component system when non preferred carbon sources (like mannitol) are utilized. Novel targets of the CbrAB/CrcZ/Crc system in P. aeruginosa were identified using transcriptome analysis; among them dctA and dctPQM were detected. CCR is regulating the dct transporter genes expression depending on the succinate concentrations in the medium of growth; this modulation of CCR is possible because, at the same time, succinate concentrations tune CCR. In a medium containing high succinate concentrations, CrcZ levels were low and therefore Crc inhibited the translation of mRNA targets. Whereas in a medium containing low succinate concentrations, the subsequent increase of CrcZ levels sequestered Crc, inhibiting its activity. This model shows for the first time that CCR possesses a feedback-based circuitry, a very important type of regulatory loop that confers the best adaptive response under changing environmental conditions. The expression of the dct transporter genes is also found to be regulated by the RNA chaperone protein Hfq. Hfq has the same post-transcriptional effect than Crc at high concentration of succinate, i.e. inhibiting dctP and dctR and indirectly favouring dctA expression. Moreover, an additional indirect positive regulation of dctP expression by Hfq was found. Finally, a metabolome approach was performed to investigate the internal signals modulating CCR via induction of CbrA activity in P. aeruginosa PAOl and P. putida KT2442. The results of the analysis are currently under study in the laboratory. - Les acides C4-dicarboxyliques font partie des sources de carbone et d'énergie préférés de P. aeruginosa, une bactérie versatile et ubiquitaire. Néanmoins, malgré leur importance, comment la présence des acides C4-dicarboxyliques dans le milieu est sentie par la bactérie et comment ils sont transportés dans la cellule chez P. aeruginosa n'étaient pas connus. De plus, peu d'informations sur ces procédés ont été répertoriées dans la littérature. Grace à notre travail, le système de transport des acides C4-dicarboxyliques (Dct) chez P. aeruginosa a pu être caractérisé. En effet, il est composé d'un nouveau système à deux composants, nommé DctB/DctD, qui régule, en combinaison avec le facteur sigma alternatif RpoN, l'expression des deux nouveaux transporteurs des acides C4-dicarboxyliques: DctA et DctPQM. L'inactivation des gènes dctA, dctB or dctD cause un défaut de croissance des souches mutantes dans un milieu minimum contenant du succinate, fumarate ou malate; confirmation de leur rôle dans le Dct. Cependant, une croissance résiduelle du mutant dctA dans ces milieux suggérerait une redondance des transporteurs d'acides Grdicarboxyliques. Une expérience de mutagenèse dans la souche AdctA, utilisant le transposon Tn5, combiné avec un criblage génétique sur la croissance dans le succinate, nous a permis d'identifier le deuxième transporteur DctPQM. DctPQM appartient à la famille des transporteurs TRAP (tripartite ATP-independent periplasmic). Un double mutant AdctAAdctPQM ne pousse pas dans du malate ou fumarate mais par contre présente une croissance résiduelle dans le succinate suggérant l'existence de transporteurs supplémentaires pour le succinate. En réalisant des expériences de compétitions nous avons démontré que le transporteur DctPQM est plus efficace que le transporteur DctA pour l'utilisation de succinate à une concentration de l'ordre du μΜ. Par contre, DctA est le transporteur le plus important pour une concentration de succinate de l'ordre du raM. Pour la première fois, deux systèmes de transport, un avec une forte- et un avec une faible-affinité (DctA et DctPQM) pour le succinate, sont coordonnés dans leur activité de transport des acides C4- dicarboxyliques, probablement contribuant à la versatilité de la bactérie. Ensuite, nous avons étudié la régulation du system Dct. En effectuant, en parallèle, une étude sur le phénomène de la répression catabolique (RC) chez P. aeruginosa, un lien entre la RC et le système Dct a été observé. La cascade des régulateurs formant la RC est composée de CbrA/CbrB, CrcZ et Crc. Crc est un répresseur traductionnel qui agit quand des sources de carbone préférées (comme les acides C4-dicarboxyliques) sont présentes dans le milieu. CrcZ est un petit ARN non-codant qui agit comme antagoniste de Crc. L'expression de CrcZ est induite par le système à deux composants CbrA/CbrB lorsque une source de carbone non-préférée est utilisée (comme le mannitol). Des nouvelles cibles du système CbrAB/CrcZ/Crc chez P. aeruginosa ont été identifiées grâce à une analyse du transcriptome des souches mutantes des régulateurs de la cascade. Parmi les cibles identifiées, les gènes dctA et dctPQM étaient présents. La RC régule l'expression des transporteurs dct en fonction de la concentration de succinate dans le milieu de croissance. Cette régulation est possible parce que, en même temps, les acides C4- dicarboxyliques régulent la RC. Dans un milieu contenant une grande concentration du succinate, le niveau d'expression de CrcZ est faible, donc Crc peut inhiber l'expression de ces ARN messagers cibles. Par contre, dans un milieu avec une faible concentration de succinate, l'augmentation de l'expression de CrcZ titre Crc et inhibe son activité. Ce modèle de régulation rétroactive est très important pour le phénomène de la RC, parce qu'il permet à la bactérie d'accorder une meilleure réponse à un changement environnemental. L'expression des gènes codant pour les transporteurs dct sont aussi régulés par la protéine chaperonne d'ARN Hfq. Hfq semble avoir le même effet traductionnelle que Crc, lorsqu'il y a une forte concentration de succinate. Nous avons ainsi observé une régulation négative de l'expression du gène dct Ρ et dctR, qui code pour un répresseur de la transcription de dctA. Nous avons aussi observé une régulation positive de la transcription de dctP par Hfq, probablement de façon indirecte. Enfin, une analyse du metabolome a était utilisée pour chercher les signaux internes modulant la RC et, en particulier, l'activité de la protéine senseur CbrA chez P. aeruginosa PAOl et P. putida KT2442. Les résultats de l'analyse sont en cours d'étude dans le laboratoire.

Alum interaction with dendritic cell membrane lipids is essential for its adjuvanticity.

As an approved vaccine adjuvant for use in humans, alum has vast health implications, but, as it is a crystal, questions remain regarding its mechanism. Furthermore, little is known about the target cells, receptors, and signaling pathways engaged by alum. Here we report that, independent of inflammasome and membrane proteins, alum binds dendritic cell (DC) plasma membrane lipids with substantial force. Subsequent lipid sorting activates an abortive phagocytic response that leads to antigen uptake. Such activated DCs, without further association with alum, show high affinity and stable binding with CD4(+) T cells via the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1). We propose that alum triggers DC responses by altering membrane lipid structures. This study therefore suggests an unexpected mechanism for how this crystalline structure interacts with the immune system and how the DC plasma membrane may behave as a general sensor for solid structures.

Subtype-specific Modulation of Acid-sensing Ion Channel (ASIC) Function by 2-Guanidine-4-methylquinazoline.

Acid-sensing ion channels (ASICs) are neuronal Na(+)-selective channels that are transiently activated by extracellular acidification. ASICs are involved in fear and anxiety, learning, neurodegeneration after ischemic stroke, and pain sensation. The small molecule 2-guanidine-4-methylquinazoline (GMQ) was recently shown to open ASIC3 at physiological pH. We have investigated the mechanisms underlying this effect and the possibility that GMQ may alter the function of other ASICs besides ASIC3. GMQ shifts the pH dependence of activation to more acidic pH in ASIC1a and ASIC1b, whereas in ASIC3 this shift goes in the opposite direction and is accompanied by a decrease in its steepness. GMQ also induces an acidic shift of the pH dependence of inactivation of ASIC1a, -1b, -2a, and -3. As a consequence, the activation and inactivation curves of ASIC3 but not other ASICs overlap in the presence of GMQ at pH 7.4, thereby creating a window current. At concentrations >1 mm, GMQ decreases maximal peak currents by reducing the unitary current amplitude. Mutation of residue Glu-79 in the palm domain of ASIC3, previously shown to be critical for channel opening by GMQ, disrupted the GMQ effects on inactivation but not activation. This suggests that this residue is involved in the consequences of GMQ binding rather than in the binding interaction itself. This study describes the mechanisms underlying the effects of a novel class of ligands that modulate the function of all ASICs as well as activate ASIC3 at physiological pH.

A 338-bp proximal fragment of the glucose transporter type 2 (GLUT2) promoter drives reporter gene expression in the pancreatic islets of transgenic mice.

The high Km glucose transporter GLUT2 is a membrane protein expressed in tissues involved in maintaining glucose homeostasis, and in cells where glucose-sensing is necessary. In many experimental models of diabetes, GLUT2 gene expression is decreased in pancreatic beta-cells, which could lead to a loss of glucose-induced insulin secretion. In order to identify factors involved in pancreatic beta-cell specific expression of GLUT2, we have recently cloned the murine GLUT2 promoter and identified cis-elements within the 338-bp of the proximal promoter capable of binding islet-specific trans-acting factors. Furthermore, in transient transfection studies, this 338-bp fragment could efficiently drive the expression of the chloramphenicol acetyl transferase (CAT) gene in cell lines derived from the endocrine pancreas, but displayed no promoter activity in non-pancreatic cells. In this report, we tested the cell-specific expression of a CAT reporter gene driven by a short (338 bp) and a larger (1311 bp) fragment of the GLUT2 promoter in transgenic mice. We generated ten transgenic lines that integrated one of the constructs. CAT mRNA expression in transgenic tissues was assessed using the RNAse protection assay and the quantitative reverse transcribed polymerase chain reaction (RT-PCR). Overall CAT mRNA expression for both constructs was low compared to endogenous GLUT2 mRNA levels but the reporter transcript could be detected in all animals in the pancreatic islets and the liver, and in a few transgenic lines in the kidney and the small intestine. The CAT protein was also present in Langerhans islets and in the liver for both constructs by immunocytochemistry. These findings suggest that the proximal 338 bp of the murine GLUT2 promoter contain cis-elements required for the islet-specific expression of GLUT2.

Glucose sensing by the hepatoportal sensor is GLUT2-dependent: in vivo analysis in GLUT2-null mice.

In the preceding article, we demonstrated that activation of the hepatoportal glucose sensor led to a paradoxical development of hypoglycemia that was associated with increased glucose utilization by a subset of tissues. In this study, we tested whether GLUT2 plays a role in the portal glucose-sensing system that is similar to its involvement in pancreatic beta-cells. Awake RIPGLUT1 x GLUT2-/- and control mice were infused with glucose through the portal (Po-) or the femoral (Fe-) vein for 3 h at a rate equivalent to the endogenous glucose production rate. Blood glucose and plasma insulin concentrations were continuously monitored. Glucose turnover, glycolysis, and glycogen synthesis rates were determined by the 3H-glucose infusion technique. We showed that portal glucose infusion in RIPGLUT1 x GLUT24-/- mice did not induce the hypoglycemia observed in control mice but, in contrast, led to a transient hyperglycemic state followed by a return to normoglycemia; this glycemic pattern was similar to that observed in control Fe-mice and RIPGLUT1 x GLUT2-/- Fe-mice. Plasma insulin profiles during the infusion period were similar in control and RIPGLUT1 x GLUT2-/- Po- and Fe-mice. The lack of hypoglycemia development in RIPGLUT1 x GLUT2-/- mice was not due to the absence of GLUT2 in the liver. Indeed, reexpression by transgenesis of this transporter in hepatocytes did not restore the development of hypoglycemia after initiating portal vein glucose infusion. In the absence of GLUT2, glucose turnover increased in Po-mice to the same extent as that in RIPGLUT1 x GLUT2-/- or control Fe-mice. Finally, co-infusion of somatostatin with glucose prevented development of hypoglycemia in control Po-mice, but it did not affect the glycemia or insulinemia of RIPGLUT1 x GLUT2-/- Po-mice. Together, our data demonstrate that GLUT2 is required for the function of the hepatoportal glucose sensor and that somatostatin could inhibit the glucose signal by interfering with GLUT2-expressing sensing units.

Spatially-resolved eigenmode decomposition of red blood cells membrane fluctuations questions the role of ATP in flickering.

Red blood cells (RBCs) present unique reversible shape deformability, essential for both function and survival, resulting notably in cell membrane fluctuations (CMF). These CMF have been subject of many studies in order to obtain a better understanding of these remarkable biomechanical membrane properties altered in some pathological states including blood diseases. In particular the discussion over the thermal or metabolic origin of the CMF has led in the past to a large number of investigations and modeling. However, the origin of the CMF is still debated. In this article, we present an analysis of the CMF of RBCs by combining digital holographic microscopy (DHM) with an orthogonal subspace decomposition of the imaging data. These subspace components can be reliably identified and quantified as the eigenmode basis of CMF that minimizes the deformation energy of the RBC structure. By fitting the observed fluctuation modes with a theoretical dynamic model, we find that the CMF are mainly governed by the bending elasticity of the membrane and that shear and tension elasticities have only a marginal influence on the membrane fluctations of the discocyte RBC. Further, our experiments show that the role of ATP as a driving force of CMF is questionable. ATP, however, seems to be required to maintain the unique biomechanical properties of the RBC membrane that lead to thermally excited CMF.

Multi-layer amniotic membrane (MLAM) transplantation for non traumatic corneal perforations or descemetoceles

Purpose:To describe the indications, the surgical procedure and the clinical outcome of MLAM in the treatment of non traumatic corneal perforations and descemetoceles . Methods:A prospective, non comparative, interventional case series of eight consecutive patients (mean age 59 years old, 6 men and 2 women) with non traumatic corneal perforations or descemetoceles.The surgery consisted in a MLAM transplantation of a cryopreservated human amniotic membrane. The series included: three active herpetic keratitis, one rosacea, one perforation of an hydrops, one cicatricial pemphigoid, one perforation after an abcess in a corneal graft and one perforation after protonbeamtherapy. The clinical outcome included: the follow-up, the integrity of the eye, corneal epithelialization, inflammation and neovascularization, and the integration of the MLAM. Stromal thickness was followed precisely with the slit lamp. A corneal graft was performed at one patient after the MLAM, allowing microscopic investigation of the removed MLAM integrated in the cornea. Results:The mean follow-up was 8.78 months (range 3.57 to 30.17). Amniotic membrane transplantation was successful and reduced inflammation in 7 patients out of 8 ,after one procedure.One patient who presented a large herpetic keratitis epithelial defect with corneal anaesthesia had his MLAM dissolved after two weeks with an aqueous leakage. Epithelium healed within 3 weeks above 7 MLAM and remained stable at 3 months in 7 out of 8 patients. MLAM opacification gradually disappeared over a few months, however, stromal layers filling in the corneal perforations or above the descemetoceles remained stable. Conclusions:MLAM transplantation is a safe, effective and useful technique to cure non traumatic corneal perforations and descemetoceles. It can be performed in emergency despite the presence of an active inflammation or infection. By facilitating epithelialization, reducing inflammation and neovascularization, it allows corneal surface reconstruction in patients with persistent epithelial defects and corneal melting that usually ends in a perforation. For full visual rehabilitation, a delayed penetrating keratoplasty is required.

Structural characterization of rockfall sources in Yosemite Valley from remote sensing data and field surveys

Yosemite Valley poses significant rockfall hazard and related risk due to its glacially steepened walls and approximately 4 million visitors annually. To assess rockfall hazard, it is necessary to evaluate the geologic structure that contributes to the destabilization of rockfall sources and locate the most probable future source areas. Coupling new remote sensing techniques (Terrestrial Laser Scanning, Aerial Laser Scanning) and traditional field surveys, we investigated the regional geologic and structural setting, the orientation of the primary discontinuity sets for large areas of Yosemite Valley, and the specific discontinuity sets present at active rockfall sources. This information, combined with better understanding of the geologic processes that contribute to the progressive destabilization and triggering of granitic rock slabs, contributes to a more accurate rockfall susceptibility assessment for Yosemite Valley and elsewhere.

Imaging pheromone sensing in a mouse vomeronasal acute tissue slice preparation.

Peter Karlson and Martin Lüscher used the term pheromone for the first time in 1959 to describe chemicals used for intra-species communication. Pheromones are volatile or non-volatile short-lived molecules secreted and/or contained in biological fluids, such as urine, a liquid known to be a main source of pheromones. Pheromonal communication is implicated in a variety of key animal modalities such as kin interactions, hierarchical organisations and sexual interactions and are consequently directly correlated with the survival of a given species. In mice, the ability to detect pheromones is principally mediated by the vomeronasal organ (VNO), a paired structure located at the base of the nasal cavity, and enclosed in a cartilaginous capsule. Each VNO has a tubular shape with a lumen allowing the contact with the external chemical world. The sensory neuroepithelium is principally composed of vomeronasal bipolar sensory neurons (VSNs). Each VSN extends a single dendrite to the lumen ending in a large dendritic knob bearing up to 100 microvilli implicated in chemical detection. Numerous subpopulations of VSNs are present. They are differentiated by the chemoreceptor they express and thus possibly by the ligand(s) they recognize. Two main vomeronasal receptor families, V1Rs and V2Rs, are composed respectively by 240 and 120 members and are expressed in separate layers of the neuroepithelium. Olfactory receptors (ORs) and formyl peptide receptors (FPRs) are also expressed in VSNs. Whether or not these neuronal subpopulations use the same downstream signalling pathway for sensing pheromones is unknown. Despite a major role played by a calcium-permeable channel (TRPC2) present in the microvilli of mature neurons TRPC2 independent transduction channels have been suggested. Due to the high number of neuronal subpopulations and the peculiar morphology of the organ, pharmacological and physiological investigations of the signalling elements present in the VNO are complex. Here, we present an acute tissue slice preparation of the mouse VNO for performing calcium imaging investigations. This physiological approach allows observations, in the natural environment of a living tissue, of general or individual subpopulations of VSNs previously loaded with Fura-2AM, a calcium dye. This method is also convenient for studying any GFP-tagged pheromone receptor and is adaptable for the use of other fluorescent calcium probes. As an example, we use here a VG mouse line, in which the translation of the pheromone V1rb2 receptor is linked to the expression of GFP by a polycistronic strategy.

Degradation of pathogen quorum-sensing molecules by soil bacteria: a preventive and curative biological control mechanism.

Abstract The plasmid pME6863, carrying the aiiA gene from the soil bacterium Bacillus sp. A24 that encodes a lactonase enzyme able to degrade N-acyl-homoserine lactones (AHLs), was introduced into the rhizosphere isolate Pseudomonas fluorescens P3. This strain is not an effective biological control agent against plant pathogens. The transformant P. fluorescens P3/pME6863 acquired the ability to degrade AHLs. In planta, P. fluorescens P3/pME6863 significantly reduced potato soft rot caused by Erwinia carotovora and crown gall of tomato caused by Agrobacterium tumefaciens to a similar level as Bacillus sp. A24. Little or no disease reduction was observed for the wild-type strain P3 carrying the vector plasmid without aiiA. Suppression of potato soft rot was observed even when the AHL-degrading P. fluorescens P3/pME6863 was applied to tubers 2 days after the pathogen, indicating that biocontrol was not only preventive but also curative. When antagonists were applied individually with the bacterial plant pathogens, biocontrol activity of the AHL degraders was greater than that observed with several Pseudomonas 2,4-diacetylphloroglucinol-producing strains and with Pseudomonas chlororaphis PCL1391, which relies on production of phenazine antibiotic for disease suppression. Phenazine production by this well characterized biological control strain P. chlororaphis PCL1391 is regulated by AHL-mediated quorum sensing. When P. chlororaphis PCL1391 was co-inoculated with P. fluorescens P3/pME6863 in a strain mixture, the AHL degrader interfered with the normally excellent ability of the antibiotic producer to suppress tomato vascular wilt caused by Fusarium oxysporum f. sp. lycopersici. Our results demonstrate AHL degradation as a novel biocontrol mechanism, but also demonstrate the potential for non-target interactions that can interfere with the biocontrol efficacy of other strains.