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PubMed2002

Suppression of NF-kappaB activation by infection with Toxoplasma gondii.

The interaction of host cells with microbial products or their invasion by pathogens frequently results in activation of the NF-kappaB family of transcription factors. The studies presented here reveal that in vivo, infection with Toxoplasma gondii results in the activation of NF-kappaB. To determine whether host cells could activate NF-kappaB in response to invasion by T. gondii, Western blots, immunofluorescence, and electrophoretic mobility shift assays were used to assess the response of host cells to infection. In these studies, infection of macrophages or fibroblasts with T. gondii did not result in the activation of NF-kappaB. In addition, the ability of lipopolysaccharide to activate NF-kappaB was impaired in cultures of macrophages infected with T. gondii. Together, these data demonstrate that invasion of cells by T. gondii does not lead to the activation of NF-kappaB and suggest that the parasite may actively interfere with the pathways that lead to NF-kappaB activation.

Active Transport, Cell Nucleus↗

PubMed2004

Inactivation of Escherichia coli O157:H7 by high-intensity ultrasonication in the presence of salts.

Inactivation of Escherichia coli O157:H7 suspended in salt solutions (Na(2)PO(4), NaCl, NaNO(3), NH(4)Cl, CaCl(2), and AlCl(3)) at concentrations ranging from 0 to 5% (w/v) by high-intensity ultrasound at two different temperature conditions (ice water bath, 40 degrees C water bath) and three ultrasonic intensity levels (9.5, 21.8, 49.2 W/cm(2)) was determined. Increases in sonication treatment time, intensity, and temperature led to increased lethality of E. coli O157:H7. However, cell lethality, as a function of solute concentration, varied depending upon type of salt. CaCl(2), NaCl, NaNO(3), Na(2)PO(4), and NH(4)Cl had little or no effect on survival of E. coli O157:H7 (<1-log reduction) regardless of other treatment conditions at 9.5 W/cm(2). Concentrations of >0.5% of CaCl(2), NaNO(3), Na(2)PO(4), and NH(4)Cl adversely affected survival (1.0- to 1.6-log reduction) with treatment of 21.8 W/cm(2) in an ice bath and showed greater inactivation (1.2-4.0-log reduction) in the 40 degrees C water bath. Treatment of 49.2 W/cm(2) showed the greatest impact regardless of other parameters. In the 40 degrees C water bath, treatment for 10 min at 49.22 W/cm(2) led to total inactivation for cells suspended in 0.5% NH(4)Cl, 1% CaCl(2), 2% NaCl, 5% Na(2)PO(4), and all concentrations of AlCl(3). Complete inactivation also occurred in an ice bath for 5% AlCl(3), which was shown to be the most effective salt. The most efficient treatment combination was in the 40 degrees C water bath for 10 min at 49.2 W/cm(2) with 5% AlCl(3). Inactivation of E. coli O157:H7 is attributed to cavitation-induced shear forces, reaction of cavitation-generated hydrogen peroxide with microbial cell wall constituents and electrostatic interactions of dissociated salts with cell membranes.

Colony Count, Microbial↗

PubMedDate not supplied

Human neutrophil antimicrobial activity.

Polymorphonuclear neutrophilic leukocytes (PMNs) take up opsonized microorganisms into phagosomes that fuse with secretory granules in the PMN cytoplasm to form phagolysosomes. Killing and digestion of microorganisms take place within phagolysosomes. Antimicrobial activities in phagolysosomes are divided into two classes. Oxygen (O2)-dependent mechanisms are expressed when PMNs undergo the "respiratory burst." An NADPH oxidase in the phagolysosome membrane is activated and reduces O2 to superoxide (O2-). O2 reduction is the first step in a series of reactions that produce toxic oxidants. For example, .O2- dismutases to hydrogen peroxide (H2O2), and the azurophil granule enzyme myeloperoxidase catalyzes the oxidation of Cl- by H2O2 to yield hypochlorous acid (HOCl). The reaction of HOCl with ammonia and amines modulates the toxicity of this oxidant. O2-independent antimicrobial mechanisms include the activities of lysosomal proteases, other hydrolytic enzymes, and proteins and peptides that bind to microorganisms and disrupt essential processes or structural components. For example, the bactericidal/permeability-increasing protein, cathepsin G, and the defensins are released into phagolysosomes from the azurophil granules. Proposed mechanisms of action of neutrophil antimicrobial agents, their range of microbial targets, and their possible interactions within phagolysosomes are discussed.

Humans↗

PubMed2026

De Novo Genome Sequence Assembly of the Algal Endosymbiont Micractinium conductrix Derived From Its Host Paramecium bursaria 186b.

Endosymbiosis is a major driver of evolutionary innovation and underpins the function of diverse ecosystems. The origins and evolution of endosymbiosis are challenging to study experimentally due to the short-lived culturability of many microbial strains derived from endosymbiotic interactions. The facultative endosymbiosis between the ciliate, Paramecium bursaria, and the green alga, Micractinium conductrix (Chlorellaceae, Trebouxiophyceae), is ecologically widespread and has emerged as a powerful lab-tractable model system. This endosymbiosis is founded upon a reciprocal nutrient exchange, but each of the species can be cultured independently enabling quantification of symbiotic fitness effects, new partnerships to be generated in the lab, and co-associations to be subject to experimental evolution. To date, evolve-and-resequence approaches have been limited due to a lack of high-quality genome assemblies enabling gene variants to be identified. Here, we report a near telomere-to-telomere genome assembly for M. conductrix 186b, using a range of sequencing technologies. Comparative analysis shows that this is one of the most complete Chlorellaceae algal genome assemblies available to date. To aid accurate gene calling and annotation, we conducted both RNAseq and Iso-Seq transcriptome sequencing experiments. Collectively, these 'omics datasets will facilitate: (i) comparative genomics studies of endosymbiont evolution, (ii) evolve-and-resequence experiments, (iii) genome-scale metabolic modeling studies, and (iv) identification of targets for genetic modification experiments and biotechnological applications.

Symbiosis↗

PubMed1998

Dendritic cells.

Dendritic cells are potent stimulators of immune responses against foreign antigens. Recent advances in this area include the delineation of distinct developmental pathways for different dendritic cell subsets; the emerging concept that one dendritic cell subset has regulatory functions that may contribute to induction of tolerance to self antigens; increased understanding of the interaction of dendritic cells with microbial products and viruses, such as HIV; and the application of dendritic cells for immunotherapy in certain cancers and possibly for the induction of transplantation tolerance.

Animals↗

PubMed2006

Methane- and sulfur-metabolizing microbial communities dominate the Lost City hydrothermal field ecosystem.

Hydrothermal venting and the formation of carbonate chimneys in the Lost City hydrothermal field (LCHF) are driven predominantly by serpentinization reactions and cooling of mantle rocks, resulting in a highly reducing, high-pH environment with abundant dissolved hydrogen and methane. Phylogenetic and terminal restriction fragment length polymorphism analyses of 16S rRNA genes in fluids and carbonate material from this site indicate the presence of organisms similar to sulfur-oxidizing, sulfate-reducing, and methane-oxidizing Bacteria as well as methanogenic and anaerobic methane-oxidizing Archaea. The presence of these metabolic groups indicates that microbial cycling of sulfur and methane may be the dominant biogeochemical processes active within this ultramafic rock-hosted environment. 16S rRNA gene sequences grouping within the Methylobacter and Thiomicrospira clades were recovered from a chemically diverse suite of carbonate chimney and fluid samples. In contrast, 16S rRNA genes corresponding to the Lost City Methanosarcinales phylotype were found exclusively in high-temperature chimneys, while a phylotype of anaerobic methanotrophic Archaea (ANME-1) was restricted to lower-temperature, less vigorously venting sites. A hyperthermophilic habitat beneath the LCHF may be reflected by 16S rRNA gene sequences belonging to Thermococcales and uncultured Crenarchaeota identified in vent fluids. The finding of a diverse microbial ecosystem supported by the interaction of high-temperature, high-pH fluids resulting from serpentinization reactions in the subsurface provides insight into the biogeochemistry of what may be a pervasive process in ultramafic subseafloor environments.

Archaea↗

PubMed2006

Cytosolic entry controls CD8+-T-cell potency during bacterial infection.

Interaction with host immunoreceptors during microbial infection directly impacts the magnitude of the ensuing innate immune response. How these signals affect the quality of the adaptive T-cell response remains poorly understood. Utilizing an engineered strain of the intracellular pathogen Listeria monocytogenes that infects cells but fails to escape from the phagosome, we demonstrate the induction of long-lived memory T cells that are capable of secondary expansion and effector function but are incapable of providing protective immunity. We demonstrate that microbial invasion of the cytosol is required for dendritic cell activation and integration of CD40 signaling, ultimately determining the ability of the elicited CD8+-T-cell pool to protect against lethal wild-type L. monocytogenes challenge. These results reveal a crucial role for phagosomal escape, not for delivery of antigen to the class I major histocompatibility complex pathway but for establishing the appropriate cellular context during CD8+-T-cell priming.

Animals↗

PubMed1988

Lipopolysaccharide structure determines ionic and hydrophobic binding of a cationic antimicrobial neutrophil granule protein.

Bactericidal activity and binding of a 57,000-dalton cationic antimicrobial neutrophil granule protein (CAP57) are determined by the presence on bacteria of O-antigen polysaccharide chains and the availability of negatively charged groups in the lipid A region, the inner core region, or both regions of lipopolysaccharide. Polymyxin B (PMB)-resistant mutants with well-defined alterations in lipid A structure and charge (pmrA) are also more resistant to CAP57. We used biologically active radioiodinated CAP57 to study the characteristics and kinetics of binding to a sensitive Rb lipopolysaccharide chemotype, Salmonella typhimurium SH9178, and the relatively resistant pmrA mutant strain SH7426. Binding occurred rapidly and was specific and saturable. Because CAP57 appears to be bound in a manner similar to that of PMB, competition binding studies were performed. Excess PMB did compete with CAP57 for binding to SH9178. Nonapeptide, a polycationic derivative of PMB that has lost its hydrophobic portions, demonstrated a marked decrease in ability to compete for binding with CAP57 compared with PMB. This demonstrated the importance of hydrophobic binding in the interaction of CAP57 with the microbial surface. Thus, we have shown that binding of CAP57 to SH9178 is specific, saturable, and similar to binding of PMB. Both hydrophobic and ionic properties of CAP57 appear to be necessary for binding.

Antimicrobial Cationic Peptides↗

PubMed2006

Regulation of physiological and pathological Th1 and Th2 responses by lactoferrin.

In recent years, Lf has gained increasing interest as a result of its protective effects against a variety of diseases. While iron binding and interactions with mammalian receptors and microbial components are the best described mechanisms of action, recent studies have provided evidence that Lf properties may be related to immunoregulatory effects on Th1/Th2 cell activities. In vitro and in vivo experiments show that Lf is able to stimulate the differentiation of T cells from their immature precursors through the induction of the CD4 antigen. Studies performed under nonpathogenic conditions have shown distinct results with regard to the ability of Lf to support the proliferation and differentiation of Th cells into the Th1 or the Th2 phenotype. In addition, Lf plays different roles in diseases by affecting the Th1/Th2 cytokine balance in a manner dependent on the host's immune status. Thus, Lf could cause a Th1 polarization in diseases in which the ability to control infection or tumor relies on a strong Th1 response. Lf may also reduce the Th1 component to limit excessive inflammatory responses. Finally, Lf may provide protection against Th1- or Th2-induced diseases, such as autoimmune or allergic diseases, through correction of the Th1/Th2 imbalance.

Administration, Oral↗

PubMed2002

Toxins of filamentous fungi.

Mycotoxins are low-molecular-weight secondary metabolites of fungi. The most significant mycotoxins are contaminants of agricultural commodities, foods and feeds. Fungi that produce these toxins do so both prior to harvest and during storage. Although contamination of commodities by toxigenic fungi occurs frequently in areas with a hot and humid climate (i.e. conditions favorable for fungal growth), they can also be found in temperate conditions. Production of mycotoxins is dependent upon the type of producing fungus and environmental conditions such as the substrate, water activity (moisture and relative humidity), duration of exposure to stress conditions and microbial, insect or other animal interactions. Although outbreaks of mycotoxicoses in humans have been documented, several of these have not been well characterized, neither has a direct correlation between the mycotoxin and resulting toxic effect been well established in vivo. Even though the specific modes of action of most of the toxins are not well established, acute and chronic effects in prokaryotic and eukaryotic systems, including humans have been reported. The toxicity of the mycotoxins varies considerably with the toxin, the animal species exposed to it, and the extent of exposure, age and nutritional status. Most of the toxic effects of mycotoxins are limited to specific organs, but several mycotoxins affect many organs. Induction of cancer by some mycotoxins is a major concern as a chronic effect of these toxins. It is nearly impossible to eliminate mycotoxins from the foods and feed in spite of the regulatory efforts at the national and international levels to remove the contaminated commodities. This is because mycotoxins are highly stable compounds, the producing fungi are ubiquitous, and food contamination can occur both before and after harvest. Nevertheless, good farm management practices and adequate storage facilities minimize the toxin contamination problems. Current research is designed to develop natural biocontrol competitive fungi and to enhance host resistance against fungal growth or toxin production. These efforts could prevent toxin formation entirely. Rigorous programs for reducing the risk of human and animal exposure to contaminated foods and feed also include economically feasible and safe detoxification processes and dietary modifications. Although risk assessment has been made for some mycotoxins, additional, systematic epidemological data for human exposure is needed for establishing toxicological parameters for mycotoxins and the safe dose for humans. It is unreasonable to expect complete elimination of the mycotoxin problem. But multiple approaches will be needed to minimize the economic impact of the toxins on the entire agriculture industry and their harmfulness to human and animal health.

Aflatoxins↗

PubMed2006

Score-based prediction of genomic islands in prokaryotic genomes using hidden Markov models.

BACKGROUND: Horizontal gene transfer (HGT) is considered a strong evolutionary force shaping the content of microbial genomes in a substantial manner. It is the difference in speed enabling the rapid adaptation to changing environmental demands that distinguishes HGT from gene genesis, duplications or mutations. For a precise characterization, algorithms are needed that identify transfer events with high reliability. Frequently, the transferred pieces of DNA have a considerable length, comprise several genes and are called genomic islands (GIs) or more specifically pathogenicity or symbiotic islands. RESULTS: We have implemented the program SIGI-HMM that predicts GIs and the putative donor of each individual alien gene. It is based on the analysis of codon usage (CU) of each individual gene of a genome under study. CU of each gene is compared against a carefully selected set of CU tables representing microbial donors or highly expressed genes. Multiple tests are used to identify putatively alien genes, to predict putative donors and to mask putatively highly expressed genes. Thus, we determine the states and emission probabilities of an inhomogeneous hidden Markov model working on gene level. For the transition probabilities, we draw upon classical test theory with the intention of integrating a sensitivity controller in a consistent manner. SIGI-HMM was written in JAVA and is publicly available. It accepts as input any file created according to the EMBL-format.It generates output in the common GFF format readable for genome browsers. Benchmark tests showed that the output of SIGI-HMM is in agreement with known findings. Its predictions were both consistent with annotated GIs and with predictions generated by different methods. CONCLUSION: SIGI-HMM is a sensitive tool for the identification of GIs in microbial genomes. It allows to interactively analyze genomes in detail and to generate or to test hypotheses about the origin of acquired genes.

Algorithms↗

PubMed1999

Association of a vitamin D receptor gene polymorphism with localized early-onset periodontal diseases.

BACKGROUND: Early-onset periodontal diseases (EOP) are caused by interactions between host factors, specific microbial pathogens, and environmental factors. It is, therefore, of interest to investigate the nature of host factors as they may provide useful risk markers and reveal important information regarding the disease pathogenesis. Genetic polymorphisms in the vitamin D receptor (VDR) gene are associated with parameters of bone homeostasis and with diseases in which bone loss is a cardinal sign, in particular osteoporosis. Rapidly progressive bone loss is one feature of EOP. We, therefore, sought to determine whether EOP is associated with a polymorphism in the VDR gene. METHODS: A restriction fragment length polymorphism (RFLP) for Taq I in exon nine of the VDR gene was analyzed by PCR, followed by restriction digestion with Taq I and gel electrophoresis. We analyzed the genotypes of 69 EOP patients, including 20 patients with unequivocal evidence of localized disease (L-EOP), and 72 controls with no history of EOP. RESULTS: The genotype distribution in the L-EOP patient group was 7 (35%), 5 (25%) and 8 (40%) and in the control group 31 (43.1%), 36 (50.0%) and 5 (6.9%) for TT, Tt and tt respectively (where t and T represent the alleles with and without the Taq I RFLP respectively). Chi2 analysis indicated that the distribution of the genotypes between these two groups was highly significantly different (P = 0.001). Allele frequencies were 47.5% and 52.5% for T and t in the L-EOP group; 68.1% and 31.9% in the control group, showing a significant association between the prevalence of the less frequent allele (t) and L-EOP (P = 0.017). There was no significant difference in the genotype distribution or the allele frequencies between the control samples and the larger EOP patient group (n = 69) which included patients with generalized and localized disease. CONCLUSIONS: These data indicate that carriage of the less frequent allele of the Taq I RFLP (t) in the VDR gene significantly increases the risk of developing L-EOP. However, VDR genotype may not affect the incidence of all cases of EOP. These findings contribute to our understanding of the genetic basis for periodontal disease and may help define sub-groups of this disease which share common pathogenic factors.

Adolescent↗

PubMed1996

Nitrification and ammonification in aquatic systems.

Nitrification is an essential step in the nitrogen cycle of natural systems because it links organic matter degradation to fixed nitrogen loss. Ammonium released by ammonification is oxidized to nitrate by nitrification, and can then be reduced to dinitrogen gas by denitrification, resulting in net loss of fixed nitrogen from the system. Whether organic matter degradation results in net ammonium release depends largely on the quality of the organic substrate and interactions among members of the microbial community involved in nitrogen and organic matter cycling. In sediments, nitrogen cycle processes depend on the supply of organic matter and oxygen from overlying water. The nature of the net flux (which direction and which form of nitrogen) is a function of closely coupled reactions (ammonification-nitrification-denitrification) in the nitrogen cycle.

Ammonia↗

PubMed1992

[The role of iron in immunologic processes].

Iron, apart a for long time well-known function connected with: transportation (hemoglobin), storage (myoglobin), and utilize (cytochromes, cytochrome oxidase) oxygen for respiration, has a critical role in host-pathogen interactions. Iron is essential for microbial growth, but also for immune function. The role of iron in infection, thermoregulation, acute lymphocytic leukemia, neoplasia, rheumatoid arthritis, stimulation of free radical reactions, and studies with iron chelation therapy are discussed.

Arthritis, Rheumatoid↗

PubMed1991

On salivary film formation and bacterial retention to solids. A methodological and experimental in vitro study.

A parallel-plate flow cell system was developed to assess individual and combined parameters relevant to oral microbial retention. In this system, internal reflection germanium prisms represented oral solid surfaces including tooth surfaces. The system used, allowed adsorbed protein films to be analyzed in situ by multiple non-destructive surface analytical techniques, without provoking surface associated biofilms. Using a range of biochemical and biophysical techniques, germanium prisms of medium critical surface tension were noted to accurately model film formation at tooth-saliva interfaces. A range of clinically relevant temperatures (22 and 37 degrees C), rinse shear rates (1 and 32 ml/min) and critical surface tensions (high, medium and low) were systematically tested in the flow-cell system for their influence, if any, on salivary microbial retention from whole human saliva. Comparisons of the interfacial organization of adsorbed material were made between human parotid (HPS) and submandibular-sublingual (HSMSL) salivas. The influence of these salivary secretions was also tested on the retention capacities of Streptococcus sanguis and Streptococcus salivarius. Microbial retention was found to be significantly dependent on initial critical surface tensions and rinse flow rates, but not to be affected by tested temperatures. When compared to prisms of low and high critical surface tension, medium critical surface tension prisms retained the highest numbers of microorganisms. No statistically verified morphological selectivity was observed in the retained microbial populations. When the interfacial organizations of HPS and HSMSL were evaluated at low- and medium energy surfaces in the absence and presence the streptococci, comparatively thinner and denser films with pronounced bacterial retention was noted for HPS on prisms of low critical surface tension. In the presence of saliva or salivary fractions, no difference in retention capacities was observed between the tested bacterial strains. The data suggest that in the presence of saliva, physico-chemical and mechanical factors have a major influence on the short term microbial retention to solid surfaces. The amount of salivary proteins being initially retained to a material also seems to be more directly related to the critical surface tension quality of the solid than to the type of bacterial strain present in suspension. The empirical critical surface tension of a material is thus a useful predictor of both microbial retention capacities and primary interactions between saliva and solid surfaces.

Bacterial Adhesion↗

PubMedDate not supplied

[Interaction of the glycoprotein from the Bacillus pumilis cell wall with liposomes].

The methods of centrifugation and gel-filtration on Sephadexes G-50 and G-150 were used to study the interaction of Bacillus pumilis cell wall glycoprotein component having the molecular weight of 50 kDa (GP-50) with lyposomes from bacterial lipids. GP-50 is shown to sorb on such liposomes and disturb their barrier properties inducing yield of low-molecular label. GP-50 exerts no effect on properties of liposomes from egg lecithin. Electrostatic forces are supposed to play a decisive role in initial acts of GP-50 interactions with lipid phase of microbial envelopes.

Bacillus↗

PubMedDate not supplied

Quantitation of Bacillus anthracis by using of soybean agglutinin conjugates.

We examine the possibility of using the soybean agglutinin (SBA) marked by peroxidase (HRP), biotin, FITC, or gold in order to determine the number of Bacillus anthracis cells of vaccine strain STI. It was shown that the technique based on interaction between the lectin and microbial cell walls likely are not inferior in sensitivity to traditional ELISA variants. The sensitivities of methods were 10(4) cells/ml in the case of SBA-biotin, 10(5) cells/ml in the case of SBA-HRP, or 10(6) cells/ml in the cases of SBA-gold and SBA-FITC.

Bacillus anthracis↗

PubMed2002

Interactions between biocide cationic agents and bacterial biofilms.

The resistance of bacterial biofilms to physical and chemical agents is attributed in the literature to various interconnected processes. The limitation of mass transfer alters the growth rate, and physiological changes in the bacteria in the film also appear. The present work describes an approach to determination of the mechanisms involved in the resistance of bacteria to quaternary ammonium compounds (benzalkonium chloride) according to the C-chain lengths of those compounds. For Pseudomonas aeruginosa CIP A 22, the level of resistance of the bacteria in the biofilm relative to that of planktonic bacteria increased with the C-chain length. For cells within the biofilm, the exopolysaccharide induced a characteristic increase in surface hydrophilicity. However, this hydrophilicity was eliminated by simple resuspension and washing. The sensitivity to quaternary ammonium compounds was restored to over 90%. Staphylococcus aureus CIP 53 154 had a very high level of resistance when it was in the biofilm form. A characteristic of bacteria from the biofilm was a reduction in the percent hydrophobicity, but the essential point is that this hydrophobicity was retained after the biofilm bacteria were resuspended and washed. The recovery of sensitivity was thus only partial. These results indicate that the factors involved in biofilm resistance to quaternary ammonium compounds vary according to the bacterial modifications induced by the formation of a biofilm. In the case of P. aeruginosa, we have underlined the involvement of the exopolysaccharide and particularly the three-dimensional structure (water channels). In the case of S. aureus, the role of the three-dimensional structure is limited and drastic physiological changes in the biofilm cells are more highly implicated in resistance.

Animals↗