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The role of Toll-like receptor 2 in microbial disease and immunity.

Cells expressing Toll-like receptor (TLR), TLR2 in association with TLR1, TLR6 or some other unknown co-receptor can respond upon interaction with a large variety of microbial ligands. The variety of TLR2 ligands is the greatest among all the TLRs and this is due to the heterodimerization needed for TLR2 mediated responses. Like other TLRs, TLR2 signaling induces antigen presenting cell activation, pro-inflammatory cytokine production and increased expression of co-stimulatory ligand expression. These events are important for induction of innate immune responses and improved acquired immunity. There is strong suggestive evidence that alteration or lack of TLR2 function in vivo may correlate to decreased immune protection from pathogens that contain TLR2 ligands, but more work needs to be performed to strengthen this correlation.

Animals↗

A new approach to non-destructive analysis of biofilms by confocal Raman microscopy.

Confocal Raman microscopy (CRM) of biofilms enables one to determine the distribution of different microorganisms and other substances inside physiological intact microbial communities. These biofilms are of outstanding interest for biological wastewater treatment. In contrast to invasive techniques, such as fluorescent in situ hybridization (FISH), we were able to identify anaerobically ammonium-oxidising (anammox) bacteria without pretreatment processes of the samples just by its Raman vibrational signature. The presented results provide new insights into the complex interactions of different organisms in microbial communities without interfering with them.

Anaerobiosis↗

Quorum sensing and motility mediate interactions between Pseudomonas aeruginosa and Agrobacterium tumefaciens in biofilm cocultures.

In the environment, multiple microbial taxa typically coexist as communities, competing for resources and, often, physically associated within biofilms. A dual-species cocultivation model has been developed by using two ubiquitous and well studied microbes Pseudomonas aeruginosa (P.a.) and Agrobacterium tumefaciens (A.t.) as a tractable system to identify molecular mechanisms that underlie multispecies microbial associations. Several factors were found to influence coculture interactions. P.a. had a distinct growth-rate advantage in cocultures, increasing its relative abundance during planktonic and biofilm growth. P.a. also demonstrated a slight quorum-sensing-dependent increase in growth yield in liquid cocultures. P.a. dominated coculture biofilms, "blanketing" or burying immature A.t. microcolonies. P.a. flagellar and type IV pili mutant strains exhibited deficient blanketing and impaired competition in coculture biofilms, whereas, in planktonic coculture, these mutations had no effect on competition. In contrast, A.t. used motility to emigrate from coculture biofilms. In both planktonic and biofilm cocultures, A.t. remained viable for extended periods of time, coexisting with its more numerous competitor. These findings reveal that quorum-sensing-regulated functions and surface motility are important microbial competition factors for P.a. and that the outcome of competition and the relative contribution of different factors to competition are strongly influenced by the environment in which they occur.

Agrobacterium tumefaciens↗

In silico encounters: harnessing metabolic modelling to understand plant-microbe interactions.

Understanding plant-microbe interactions is vital for developing sustainable agricultural practices and mitigating the consequences of climate change on food security. Plant-microbe interactions can improve nutrient acquisition, reduce dependency on chemical fertilizers, affect plant health, growth, and yield, and impact plants' resistance to biotic and abiotic stresses. These interactions are largely driven by metabolic exchanges and can thus be understood through metabolic network modelling. Recent developments in genomics, metagenomics, phenotyping, and synthetic biology now enable researchers to harness the potential of metabolic modelling at the genome scale. Here, we review studies that utilize genome-scale metabolic modelling to study plant-microbe interactions in symbiotic, pathogenic, and microbial community systems. This review catalogues how metabolic modelling has advanced our understanding of the plant host and its associated microorganisms as a holobiont. We showcase how these models can contextualize heterogeneous datasets and serve as valuable tools to dissect and quantify underlying mechanisms. Finally, we consider studies that employ metabolic models as a testbed for in silico design of synthetic microbial communities with predefined traits. We conclude by discussing broader implications of the presented studies, future perspectives, and outstanding challenges.

Plants↗

[Influence of foods on the absorption of antimicrobial agents].

Understanding the interaction between foods and antimicrobial agents is an aspect of therapy which may have an important clinical repercussion, which is why it must not be forgotten. This study reviews the interactions of foods with the anti-microbial agents which occur at the level of absorption, considering the mechanisms involved. The food-drug interaction can cause an increase, a decrease, or a delay in the bio-availability of the anti-microbial agents; foods may have no affect on the absorption of the anti-microbial agents, or they may improve the gastrointestinal tolerance. The Food and Drug Administration's bioequivalency criteria for considering whether or not there is an alteration in absorption have an orientative function, as generally all studies are conducted on healthy volunteers, but in clinical practice one must consider the physiological and pathological condition of the patient. The composition of the diet as well as the volume of liquid administered are other aspects which should be considered, as these may exert a different effect depending on the type of drug. After ingesting fatty foods, there is an increased absorption of albendazole, griseofulvin, itraconazole, and mebendazole. All foods but especially carbohydrates reduce the absorption of isoniazide. Among the anti-HIV drugs, the following must be administered on an empty stomach: didanosine, indinavir zalcitabine , and zidovudine ; lamivudine can be administered either on an empty or on a full stomach, because although food delays the absorption, it does not affect the amount absorbed; the absorption of stavudine is not affected by foods; ritonavir should be administered together with tile meals, and saquinavir must be administered after ingestion of food. It is advisable to administer clarithromycin together with foods, and azythromycin on an empty stomach; the same holds true for perfloxacin and rifabutine as for lamivudine.

Absorption↗

Effect of surface hydrophobicity on the adhesion of S. cerevisiae onto modified surfaces by poly(styrene-ran-sulfonic acid) random copolymers.

The hydrophobicity of solid surfaces has been regarded as a controlling factor in microbial adhesion phenomena. In this study, the surface hydrophobicity was modified by coating with a poly(styrene-ran-sulfonic acid) random copolymer (PS-x-SA, charge density (x): 0-15.3%), and the adhesion rate, J0, of S. cerevisiae performed with a direct observation technique. The results indicated that the degree of sulfonation of PS-x-SA greatly influenced the hydrophobicity of substrates and the adhesion of yeast cells. The J0 of PS-x-SA substrates were gradually decreased as increasing charge density. The interactions between cells and substrates explained by the XDLVO theory, predicted that the decrease of J0 as increasing charge density was not due to the increase of electric double layer repulsion, but mainly due to the hydrophobic acid-base interactions. Also, it predicted that microbial adhesions of PS-x-SA were mostly reversible, while some of PS and PS-5.1-SA adhered cells were hardly removed. Based on these results, XDLVO theory was effective for predicting adhesion phenomena of S. cerevisiae onto the PS-x-SA-coated substrates.

Cell Adhesion↗

Persistent Chlamydia trachomatis infections resist apoptotic stimuli.

Microbial modulation of apoptosis has added a new dimension of understanding to the dynamic interaction between the human host and its microbial invaders. Persistent infection can be a by-product of inhibition of apoptosis and may significantly impact the pathogenesis of diseases caused by organisms such as Chlamydia trachomatis. We compared apoptotic responses among HeLa 229 cells acutely and persistently infected and mock infected with serovar A/HAR-13. Persistence was induced by gamma interferon at 0.2 and 2.0 ng/ml. Cells were treated with etoposide or staurosporine at 24-h intervals and assayed for apoptosis by cell count, DNA ladder formation, and cytochrome c translocation. From the 24- to 120-h time points, infected cultures were 87 and 90% viable for etoposide and staurosporine treatment, respectively, and produced no DNA ladder, and cytochrome c remained in the mitochondria. In contrast, mock-infected cells were 22 and 37% viable for etoposide (P = 0.0001) and staurosporine (P = 0.01), respectively, and displayed characteristic DNA ladders, and cytochrome c was translocated. We found that resistance to apoptotic stimuli was identical in acute and persistent infections. Since cytochrome c was not translocated from the mitochondrion, caspase-9 activity was likely not involved. The expression of chlamydial hsp60, a known stimulator of inflammation in vivo, was measured in both active and persistent infections by Western blot, with increased production in the latter with or without staurosporine treatment. Chlamydial disregulation of apoptosis and the ensuing persistence of organisms offer an alternative pathogenic mechanism for chlamydial scarring observed in trachoma and infertility populations via sustained inflammation induced by immunoreactive molecules such as hsp60.

Apoptosis↗

Propranolol metabolism by Cunninghamella bainieri.

1. Incubations of racemic propranolol alone or in the presence of either quinidine or sparteine were performed with Cunninghamella bainieri. 2. Five mammalian metabolites of propranolol (4-hydroxypropranolol, desisopropyl-propranolol, 1-naphthoxylactic acid, propranolol glycol and 1-naphthoxyacetic acid) were present in unhydrolysed extracts of the incubation medium according to h.p.l.c. and g.l.c. analyses. The relative proportion of 4-hydroxypropranolol increased after enzymic treatment. 3. Propranolol not only had a fungistatic effect, but also caused morphological changes in the organism, which were accompanied by decomposition of 4-hydroxypropranolol and formation of a greenish-brown colour in the incubation medium. 4. Drug interaction experiments yielded results which paralleled those reported in mammals. 5. The findings indicate that C. bainieri may be a useful microbial model for drug disposition and interaction studies.

Biotransformation↗

Focus on toxicological aspects of pesticide chemical interaction in drinking water contamination.

Toxicological aspects related to chemical and microbial degradation of pesticides in water and to products deriving from the interaction with xenobiotics found in water are reviewed. Other aspects considered are those related to compounds formed in potabilization processes and to water contamination by pesticide synthesis by-products or intermediates. These problems refer to scarcely investigated issues which are nevertheless very interesting because of their impact on human health.

Pesticide Residues↗

Application of atomic force microscopy to microbial surfaces: from reconstituted cell surface layers to living cells.

The application of atomic force microscopy (AFM) to probe the ultrastructure and physical properties of microbial cell surfaces is reviewed. The unique capabilities of AFM can be summarized as follows: imaging surface topography with (sub)nanometer lateral resolution; examining biological specimens under physiological conditions; measuring local properties and interaction forces. AFM is being used increasingly for: (i) visualizing the surface ultrastructure of microbial cell surface layers, including bacterial S-layers, purple membranes, porin OmpF crystals and fungal rodlet layers; (ii) monitoring conformational changes of individual membrane proteins; (iii) examining the morphology of bacterial biofilms, (iv) revealing the nanoscale structure of living microbial cells, including fungi, yeasts and bacteria, (v) mapping interaction forces at microbial surfaces, such as van der Waals and electrostatic forces, solvation forces, and steric/bridging forces; and (vi) probing the local mechanical properties of cell surface layers and of single cells.

Bacteria↗

[Microbial ecology of the colon].

It has been known for a long time that the human gastrointestinal tract contains 10(14) micro-organisms, which are predominantly anaerobic. Recent research has provided a better understanding of the functions and the equilibrium of the intestinal ecosystem in which the intestinal mucosa and the microbial flora which it supports interact. Any modification in one or other of the constituents of this ecosystem is likely to disturb the normal ecological equilibrium, resulting in a variety of gastro-intestinal diseases. Today, the intestinal ecosystem can be considered to be a system of defence and equilibrium or, conversely, as a reservoir of infection, which is confirmed by the study of certain faecal bacterial profiles with a high risk of infection. In gastrointestinal surgery and paediatrics, the correlation between antibiotic therapy, intestinal microbial proliferation and the development of septicaemia confers a fundamental role of the defence barriers of the gastrointestinal tract in the control or potentially pathogenic endogenous micro-organisms.

Colon↗

The role of interactions, sessile growth and nutrient amendments on the degradative efficiency of a microbial consortium.

A degradative microbial consortium consisting of at least nine bacterial and one algal species was isolated from soil with diclofop methyl as the sole carbon source. In continuous flow culture, the presence of the algae increased diclofop methyl degradation and removal by 36%. Batch culture experiments with 14C-labeled diclofop methyl confirmed algal involvement in the mineralization of diclofop methyl as there was no significant difference in the amount of 14CO2 evolved by the bacterial consortium with and without the algal activity when the consortium was cultivated in the dark to inhibit algal growth, while 11% more 14CO2 was produced in the light by the algal-bacterial consortium. Pure cultures isolated from the bacterial consortium could not individually mineralize diclofop methyl as the sole carbon source. However, when supplied with an additional carbon source, two strains could mineralize diclofop methyl. Addition of either the complex growth medium, or a cell-free filtrate from the algal-bacterial consortium to batch systems containing 14C-labeled diclofop methyl resulted in a significant increase in the production of 14CO2 by the bacterial consortium, suggesting co-metabolism of diclofop methyl in the presence of a labile carbon source. Removal of diclofop methyl by the bacterial consortium was increased by 36% when a larger surface to volume ratio was provided by glass beads that allowed extensive biofilm formation. The requirement for exogenous carbon sources and the inability of isolated pure cultures to degrade diclofop methyl indicated that interspecies interactions are necessary for degradation. The positive effect of sessile growth suggested that spatial organization of cells may also be important for degradation.

Bacteria↗

The University of Minnesota Biocatalysis/Biodegradation Database: post-genomic data mining.

The University of Minnesota Biocatalysis/Biodegradation Database (UM-BBD, http://umbbd.ahc.umn.edu/) provides curated information on microbial catabolism and related biotransformations, primarily for environmental pollutants. Currently, it contains information on over 130 metabolic pathways, 800 reactions, 750 compounds and 500 enzymes. In the past two years, it has increased its breath to include more examples of microbial metabolism of metals and metalloids; and expanded the types of information it includes to contain microbial biotransformations of, and binding interactions with many chemical elements. It has also increased the ways in which this data can be accessed (mined). Structure-based searching was added, for exact matches, similarity, or substructures. Analysis of UM-BBD reactions has lead to a prototype, guided, pathway prediction system. Guided prediction means that the user is shown all possible biotransformations at each step and guides the process to its conclusion. Mining the UM-BBD's data provides a unique view into how the microbial world recycles organic functional groups. UM-BBD users are encouraged to comment on all aspects of the database, including the information it contains and the tools by which it can be mined. The database and prediction system develop under the direction of the scientific community.

Biodegradation, Environmental↗

Community genomics among stratified microbial assemblages in the ocean's interior.

Microbial life predominates in the ocean, yet little is known about its genomic variability, especially along the depth continuum. We report here genomic analyses of planktonic microbial communities in the North Pacific Subtropical Gyre, from the ocean's surface to near-sea floor depths. Sequence variation in microbial community genes reflected vertical zonation of taxonomic groups, functional gene repertoires, and metabolic potential. The distributional patterns of microbial genes suggested depth-variable community trends in carbon and energy metabolism, attachment and motility, gene mobility, and host-viral interactions. Comparative genomic analyses of stratified microbial communities have the potential to provide significant insight into higher-order community organization and dynamics.

Amino Acid Sequence↗

Innate defences against methicillin-resistant Staphylococcus aureus (MRSA) infection.

The innate immune system is the primary defence against bacterial infection. Among the factors involved in innate defence, anti-microbial peptides produced by humans have recently attracted attention due to their relevance to some diseases and also to the development of new chemotherapeutic agents. Staphylococcus aureus is one of the major human pathogens, causing a variety of infections from suppurative disease to food poisoning. Methicillin-resistant S. aureus (MRSA) is a clinical problem and with the recent emergence of a vancomycin-resistant strain, this will pose serious problems in the near future. In investigating the molecular biology of S. aureus infections to develop new chemotherapeutic agents against MRSA infections, knowledge of the interaction of innate anti-microbial peptides with S. aureus is important. In vitro and in vivo experiments demonstrate that exposure of S. aureus to host cells can induce the anti-microbial peptides beta-defensin-2 (hBD2), hBD3, and LL37/CAP18. The induction level of these peptides differs among strains, as does the susceptibility of the strains, with MRSA strains exhibiting lower susceptibility. In summary, the susceptibility of S. aureus strains, including MRSA strains, to components of the innate immune system varies, with the MRSA strains showing more resistance to both innate immune factors and chemotherapeutic agents.

Anti-Bacterial Agents↗

Recognition and response in the plant immune system.

Molecular communication between plants and potential pathogens determines the ultimate outcome of their interaction. The directed delivery of microbial molecules into and around the host cell, and the subsequent perception of these by the invaded plant tissue (or lack thereof), determines the difference between disease and disease resistance. In theory, any foreign molecule produced by an invading pathogen could act as an elicitor of the broad physiological and transcriptional re-programming indicative of a plant defense response. The diversity of elicitors recognized by plants seems to support this hypothesis. Additionally, these elicitors are often virulence factors from the pathogen recognized by the host. This recognition, though genetically as simple as a ligand-receptor interaction, may require additional host proteins that are the nominal targets of virulence factor action. Transduction of recognition probably requires regulated protein degradation and results in massive changes in cellular homeostasis, including a programmed cell death known as the hypersensitive response that indicates a successful, if perhaps over-zealous, disease resistance response.

Gene Expression Regulation, Plant↗

Correlations of cationic charges with salt sensitivity and microbial specificity of cystine-stabilized beta -strand antimicrobial peptides.

The electrostatic interaction of the charge cluster of an amphipathic peptide antibiotic with microbial membranes is a salt-sensitive step that often determines organism specificity. We have examined the correlation between charge clusters and salt insensitivity and microbial specificity in linear, cyclic, and retro-isomeric cystine-stabilized beta-strand (CSbeta) tachyplesin (TP) in a panel of 10 test organisms. Cyclic tachyplesins consisting of 14 and 18 amino acids are constrained by an end-to-end peptide backbone and two or three disulfide bonds to cross-brace the anti-parallel beta-strand that approximates a "beta-tile" structure. Circular dichroism measurements of beta-tile TPs showed that they displayed ordered structures. Control peptides containing the same number of basic amino acids as TP but lacking disulfide constraints were highly salt sensitive. Cyclic TP analogues with six cationic charges were more broadly active and salt-insensitive than those with fewer cationic charges. Reducing their proximity or number of cationic charges, particularly those with three or fewer basic amino acids, led to a significant decrease in potency and salt insensitivity, but an increased selectivity to certain Gram-positive bacteria. An end-group effect of the dibasic N-terminal Lys of TP in the open-chain TP and its retroisomer was observed in certain Gram-negative bacteria under high-salt conditions, an effect that was not found in the cyclic analogs. These results suggest that a stable folded structure together with three or more basic amino acids closely packed in a charged region in CSbeta peptides is important for salt insensitivity and organism specificity.

Amino Acid Sequence↗

Helicobacter pylori virulence factors--one part of a big picture.

CONTEXT: At least half the world's population is infected with Helicobacter pylori, although only 10-20% of carriers develop gastric diseases, ranging from ulcer to MALT-lymphoma and adenocarcinoma (MALT is mucosa-associated lymphoid tissue). The clinical outcome of H pylori infection is determined by a complex interaction of environmental influences and host and microbial virulence factors. H pylori genotypes carrying the babA2 gene, encoding a bacterial adhesin mediating interaction with gastric epithelial cells, have enhanced pathogenicity. Moreover, coexistence of babA2 with other bacterial virulence factors further worsens clinical outcomes. STARTING POINT: To further elucidate the clinical relevance of babA2-genopositive H pylori strains, Carlo-Frederico Zambon and colleagues analysed the association of babA2 genotypes with gastritis, gastroduodenal ulcer disease, or intestinal metaplasia in 167 infected Italian individuals. The coexistence of babA2 with other potentially disease-related H pylori genes, such as cagA, vacA, or oipA, correlated with clinical outcome. 36% of H pylori strains were babA2(-) genopositive, and abundance of babA2 was associated with the genomic presence of the other potential virulence-factor genes. H pylori strains carrying babA2, cagA, and the vacA genotype s1m1 were associated with the highest risk of developing intestinal metaplasia, whereas this condition was rarely (<10%) associated with strains with a cagA-, babA2-, vacA s2m2 genotype. Whilst the risk of developing more serious gastric lesions increased as the number of virulence factor genes accumulated in a given H pylori strain, there was no indication of any one specific bacterial gene-pattern being associated with a particular clinical disease. WHERE NEXT? Identifying the factors responsible for the enhanced pathogenicity of H pylori leading to development of life-threatening diseases in a subset of infected individuals is a mandatory task for the future. Identification of virulence-associated H pylori genes and investigation of their clinical relevance in large prospective studies will help to define such strains with increased pathogenicity. The value of H pylori genotypes as predictors of disease outcome is limited, because the pathogenic impact of bacterial virulence factors is greatly influenced by coexisting environmental and host factors.

Genotype↗