Home About us Contact
|
Home About us Contact | ||
|
|
||
E. Coli (e + coli)
Kinds of E. Coli Terms modified by E. Coli Selected AbstractsFood Safety Regulation and the Conflict of Interest: The Case of MeatSafety and E. Coli 0157PUBLIC ADMINISTRATION, Issue 3 2000Richard Schofield The Food Standards Agency (FSA) aims to remove the longstanding conflict of interest between producers and consumers which is thought to lie at the heart of the rising number of food safety problems of recent years, to restore consumer confidence, and to protect public health. This paper sets out firstly to understand what the conflicts are, how they arise and their implications for food safety, and secondly to provide some means of evaluating the proposals for the Food Standards Agency. It does this by examining the current food safety regulatory regime as it relates to e. coli 0157, one of the problems that gave rise to the FSA and an exemplar of the problems of meat safety, and places it in its wider economic context. The results show that the financial pressures on the food industry were such that food hygiene was largely dependent upon external regulation and enforcement. But the deficiencies in the conception, design and implementation of the Food Safety Act, which was fundamentally deregulatory and privileged producer interests, permitted the food safety problems to grow. The case also, by illustrating how the interests of big business predominate in the formulation of public policy at the expense of the public, reveals how the class nature of the state affects public policy and social relations. Without addressing these issues, the problems they give rise to will remain. While the case is based on experiences in Britain, the problem of food safety and the issues raised have an international significance. [source] Design, Synthesis, and Pharmacological Investigation of Iodined Salicylimines, New Prototypes of Antimicrobial Drug CandidatesARCHIV DER PHARMAZIE, Issue 5 2010Suo-Ping Xu Abstract A series of 3,5-diiodo-salicylalidene Schiff bases (compounds 1,35) has been synthesized and tested for antimicrobial activity. The compounds were assayed for antibacterial activities by the MTT method. Some of the compounds inhibit the growth of a broad range of bacteria including the species of Bacillus subtilis, Staphylococcus aureus, Streptococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, and Enterobacter cloacae. Among them, compounds 2-[(4-chloro-phenylimino)methyl]-4,6-diiodo-phenol 11 and 2,4-diiodo-6-[(2-morpholin-4-yl-ethylimino)methyl]phenol 19 showed the most potent antibacterial activity with MIC of 3.1, 12.9, 3.3, 6.5, 12.9, 3.3 and 3.2, 12.8, 3.2, 12.8, 12.8, 3.2 ,M against B. subtilis, S. aureus, S. faecalis, P. aeruginosa, E. Coli, and E. cloacae, respectively. [source] Gastrointestinal Bacterial Transmission among Humans, Mountain Gorillas, and Livestock in Bwindi Impenetrable National Park, UgandaCONSERVATION BIOLOGY, Issue 6 2008INNOCENT B. RWEGO ecología de enfermedades; Escherichia coli; primates; salud del ecosistema; zoonosis Abstract:,Habitat overlap can increase the risks of anthroponotic and zoonotic pathogen transmission between humans, livestock, and wild apes. We collected Escherichia coli bacteria from humans, livestock, and mountain gorillas (Gorilla gorilla beringei) in Bwindi Impenetrable National Park, Uganda, from May to August 2005 to examine whether habitat overlap influences rates and patterns of pathogen transmission between humans and apes and whether livestock might facilitate transmission. We genotyped 496 E. coli isolates with repetitive extragenic palindromic polymerase chain reaction fingerprinting and measured susceptibility to 11 antibiotics with the disc-diffusion method. We conducted population genetic analyses to examine genetic differences among populations of bacteria from different hosts and locations. Gorilla populations that overlapped in their use of habitat at high rates with people and livestock harbored E. coli that were genetically similar to E. coli from those people and livestock, whereas E. coli from gorillas that did not overlap in their use of habitats with people and livestock were more distantly related to human or livestock bacteria. Thirty-five percent of isolates from humans, 27% of isolates from livestock, and 17% of isolates from gorillas were clinically resistant to at least one antibiotic used by local people, and the proportion of individual gorillas harboring resistant isolates declined across populations in proportion to decreasing degrees of habitat overlap with humans. These patterns of genetic similarity and antibiotic resistance among E. coli from populations of apes, humans, and livestock indicate that habitat overlap between species affects the dynamics of gastrointestinal bacterial transmission, perhaps through domestic animal intermediates and the physical environment. Limiting such transmission would benefit human and domestic animal health and ape conservation. Resumen:,El traslape de hábitats puede incrementar los riesgos de transmisión de patógenos antroponótica y zoonótica entre humanos, ganado y simios silvestres. Recolectamos bacterias Escherichia coli de humanos, ganado y gorilas de montaña (Gorilla gorilla beringei) en el Parque Nacional Bwindi Impenetrable, Uganda, de mayo a agosto 2005 para examinar sí el traslape de hábitat influye en las tasas y patrones de transmisión de patógenos entre humanos y simios y sí el ganado facilita esa transmisión. Determinamos el genotipo de 496 aislados de E. coli con marcaje de reacción en cadena de polimerasa palindrómica extragénica (rep-PCR) y medimos la susceptibilidad a 11 antibióticos con el método de difusión de disco. Realizamos análisis de genética poblacional para examinar las diferencias genéticas entre poblaciones de bacterias de huéspedes y localidades diferentes. Las poblaciones de gorilas con alto grado de traslape en el uso de hábitat con humanos y ganado presentaron E. coli genéticamente similar a E. coli de humanos y ganado, mientras que E. coli de gorilas sin traslape en el uso hábitat con humanos y ganado tuvo relación lejana con las bacterias de humanos y ganado. Treinta y cinco porciento de los aislados de humanos, 27% de los aislados de ganado y 17% de los aislados de gorilas fueron clínicamente resistentes a por lo menos un antibiótico utilizado por habitantes locales, y la proporción de gorilas individuales con presencia de aislados resistentes declinó en las poblaciones proporcionalmente con la disminución en el grado de traslape con humanos. Estos de patrones de similitud genética y resistencia a antibióticos entre E. coli de poblaciones de simios, humanos y ganado indican que el traslape de hábitat entre especies afecta la dinámica de transmisión de bacterias gastrointestinales, probablemente a través de animales domésticos intermediarios y el ambiente físico. La limitación de esa transmisión beneficiaría a la salud de humanos y animales domésticos y a la conservación de simios. [source] Assessment of myosin II, Va, VI and VIIa loss of function on endocytosis and endocytic vesicle motility in bone marrow-derived dendritic cellsCYTOSKELETON, Issue 10 2007Jeffrey P. Holt Abstract An essential feature of dendritic cell immune surveillance is endocytic sampling of the environment for non-self antigens primarily via macropinocytosis and phagocytosis. The role of several members of the myosin family of actin based molecular motors in dendritic cell endocytosis and endocytic vesicle movement was assessed through analysis of dendritic cells derived from mice with functionally null myosin mutations. These include the dilute (myosin Va), Snell's waltzer (myosin VI) and shaker-1 (myosin VIIa) mouse lines. Non muscle myosin II function was assessed by treatment with the inhibitor, blebbistatin. Flow cytometric analysis of dextran uptake by dendritic cells revealed that macropinocytosis was enhanced in Snell's waltzer dendritic cells while shaker-1 and blebbistatin-treated cells were comparable to controls. Comparison of fluid phase uptake using pH insensitive versus pH sensitive fluorescent dextrans revealed that in dilute cells rates of uptake were normal but endosomal acidification was accelerated. Phagocytosis, as quantified by uptake of E. coli, was normal in dilute while dendritic cells from Snell's waltzer, shaker-1 and blebbistatin treated cells exhibited decreased uptake. Microtubule mediated movements of dextran-or transferrin-tagged endocytic vesicles were significantly faster in dendritic cells lacking myosin Va. Loss of myosin II, VI or VIIa function had no significant effects on ratesof endocytic vesicle movement. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source] Models of white matter injury: Comparison of infectious, hypoxic-ischemic, and excitotoxic insultsDEVELOPMENTAL DISABILITIES RESEARCH REVIEW, Issue 1 2002Henrik Hagberg Abstract White matter damage (WMD) in preterm neonates is strongly associated with adverse outcome. The etiology of white matter injury is not known but clinical data suggest that ischemia-reperfusion and/or infection-inflammation are important factors. Furthermore, antenatal infection seems to be an important risk factor for brain injury in term infants. In order to explore the pathophysiological mechanisms of WMD and to better understand how infectious agents may affect the vulnerability of the immature brain to injury, numerous novel animal models have been developed over the past decade. WMD can be induced by antenatal or postnatal administration of microbes (E. coli or Gardnerella vaginalis), virus (border disease virus) or bacterial products (lipopolysaccharide, LPS). Alternatively, various hypoperfusion paradigms or administration of excitatory amino acid receptor agonists (excitotoxicity models) can be used. Irrespective of which insult is utilized, the maturational age of the CNS and choice of species seem critical. Generally, lesions with similarity to human WMD, with respect to distribution and morphological characteristics, are easier to induce in gyrencephalic species (rabbits, dogs, cats and sheep) than in rodents. Recently, however, models have been developed in rats (PND 1,7), using either bilateral carotid occlusion or combined hypoxia-ischemia, that produce predominantly white matter lesions. LPS is the infectious agent most often used to produce WMD in immature dogs, cats, or fetal sheep. The mechanism whereby LPS induces brain injury is not completely understood but involves activation of toll-like receptor 4 on immune cells with initiation of a generalized inflammatory response resulting in systemic hypoglycemia, perturbation of coagulation, cerebral hypoperfusion, and activation of inflammatory cells in the CNS. LPS and umbilical cord occlusion both produce WMD with quite similar distribution in 65% gestational sheep. The morphological appearance is different, however, with a more pronounced infiltration of inflammatory cells into the brain and focal microglia/macrophage ("inflammatory WMD") in response to LPS compared to hypoperfusion evoking a more diffuse microglial response usually devoid of cellular infiltrates ("ischemic WMD"). Furthermore, low doses of LPS that by themselves have no adverse effects in 7-day-old rats (maturation corresponding to the near term human fetus), dramatically increase brain injury to a subsequent hypoxic-ischemic challenge, implicating that bacterial products can sensitize the immature CNS. Contrary to this finding, other bacterial agents like lipoteichoic acid were recently shown to induce tolerance of the immature brain suggesting that the innate immune system may respond differently to various ligands, which needs to be further explored. MRDD Research Reviews 2002;8:30,38. © 2002 Wiley-Liss, Inc. [source] Dopamine and sensory tissue development in Drosophila melanogasterDEVELOPMENTAL NEUROBIOLOGY, Issue 4 2001Wendi Neckameyer Abstract Dopamine is an important signaling molecule in the nervous system; it also plays a vital role in the development of diverse non-neuronal tissues in the fruit fly Drosophila melanogaster. The current study demonstrates that males depleted of dopamine as third instar larvae (via inhibition of the biosynthetic enzyme tyrosine hydroxylase) demonstrated abnormalities in courtship behavior as adults. These defects were suggestive of abnormalities in sensory perception and/or processing. Electroretinograms (ERGs) of eyes from adults depleted of dopamine for 1 day as third instar larvae revealed diminished or absent on- and off-transients. These sensory defects were rescued by the addition of L -DOPA in conjunction with tyrosine hydroxylase inhibition during the larval stage. Depletion of dopamine in the first or second larval instar was lethal, but this was not due to a general inhibition of proliferative cells. To establish that dopamine was synthesized in tissues destined to become part of the adult sensory apparatus, transgenic lines were generated containing 1 or 4 kb of 5, upstream sequences from the Drosophila tyrosine hydroxylase gene (DTH) fused to the E. coli ,-galactosidase reporter. The DTH promoters directed expression of the reporter gene in discrete and consistent patterns within the imaginal discs, in addition to the expected expression in gonadal, brain, and cuticular tissues. The ,-galactosidase expression colocalized with tyrosine hydroxylase protein. These results are consistent with a developmental requirement for dopamine in the normal physiology of adult sensory tissues. © 2001 John Wiley & Sons, Inc. J Neurobiol 47: 280,294, 2001 [source] Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGEELECTROPHORESIS, Issue 17 2009Few Ne Chew Abstract Mechanical and non-mechanical breakages of bacterial cells are usually the preliminary steps in intracellular protein purification. In this study, the recombinant green fluorescent protein (GFP) was purified from intact Escherichia coli cells using preparative PAGE. In this purification process, cells disruption step is not needed. The cellular content of E. coli was drifted out electrically from cells and the negatively charged GFP was further electroeluted from polyacrylamide gel column. SEM investigation of the electrophoresed cells revealed substantial structural damage at the cellular level. This integrated purification technique has successfully recovered the intracellular GFP with a yield of 82% and purity of 95%. [source] On-line concentration of proteins by SDS-CGE with LIF detectionELECTROPHORESIS, Issue 2 2008Cheng-Ju Yu Abstract We present a simple approach for on-line concentration of SDS-protein complexes by using poly(vinyl alcohol) (PVA) solution in CGE. In comparison to the coated capillary, the presence of EOF in CGE omitted the need to fill the capillaries with polymer solutions prior to the analysis. More importantly, we found that highly reproducible separation of eight proteins by 3.5% PVA was achieved between runs and without the regeneration of high bulk EOF; the RSD of migration times was less than 0.7%. To further improve the concentration sensitivity, neutral PVA was introduced into the capillary with the help of EOF to act as sieving matrix. The occurrence of stacking at the boundary between the PVA and the sample zone is mainly due to the retardation of proteins by PVA. As a result, the LODs at an S/N of 3 for SDS,protein complexes are of the order of sub-nM to several nM. For example, the LOD for BSA is 0.78 nM, which is a 91-fold sensitivity enhancement over the normal injection. In addition, our stacking method has been applied to the analyses of proteins in Escherichia coli cells. The peak for ,-galactosidase (E. coli) was observed after 0.1 ,M ,-galactosidase was spiked into the E. coli samples. [source] Narrow-band fractionation of proteins from whole cell lysates using isoelectric membrane focusing and nonporous reversed-phase separationsELECTROPHORESIS, Issue 7-8 2004Yi Zhu Abstract Preparative isoelectric focusing (PIEF) is used to achieve narrow-band fractionation of proteins from whole cell lysates of Escherichia coli (E. coli). Isoelectric membranes create well-defined pH ranges that fractionate proteins by isoelectric point (pI) upon application of an electric potential. A commercial IsoPrime device (Amersham-Pharmacia BioTech) is modified for the PIEF separation to lessen run volumes significantly. Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis of chamber contents indicates that excellent pH fractionation is achieved with little overlap between chambers. PIEF pH fractions are further separated using nonporous reversed-phase high-performance liquid chromatography (NPS-RP-HPLC) and HPLC eluent is analyzed on-line by electrospray ionization-time of flight-mass spectrometry (ESI-TOF-MS) for intact protein molecular weight (MW) analysis. The result is a pI versus MW map of bacterial protein content. IEF fractionation down to 0.1 pH units combined with intact protein MW values result in a highly reproducible map that can be used for comparative analysis of different E. coli strains. [source] Enhancement of the NAD(P)(H) Pool in Escherichia coli for BiotransformationENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 4 2007F. Heuser Abstract In pyridine nucleotide-dependent, reductive whole cell biotransformation with resting cells of Escherichia coli, the availability of intracellular NAD(P)(H) is a pivotal point for an efficient and highly productive substrate conversion. The question whether an increase of the intracellular NAD(P)(H) concentration could increase the productivity was discussed controversially in the past. This is the first report on an E. coli strain with an increased NAD(P)(H) pool which was tested in a reductive biotransformation system for an increased productivity. Biotransformation was performed with a strain overexpressing a gene encoding an (R)-specific alcohol dehydrogenase for the stereospecific, NADPH-dependent reduction of methyl acetoacetate (MAA) to (R)-methyl-3-hydroxybutanoate (MHB). Cofactor regeneration was implemented via glucose oxidation by coexpression of a gene encoding glucose dehydrogenase. The specific MHB productivity (mmol mg,1 cell dry weight,1h,1) enabled a comparison between the E. coli,BL21(DE3) wild-type and a genetically modified strain. The enhancement of the NAD(P)(H) pool was achieved by genetic manipulation of the NAD(H) biosynthetic pathways. After simultaneous overexpression of the pncB and nadE genes, encoding nicotinic acid phosphoribosyltransferase and NAD synthetase, measurements of the total NAD(P)(H) pool, sizes showed a 7-fold and 2-fold increased intracellular concentration of NAD(H) and NADP(H), respectively. However, the implementation of an E.,coli strain carrying a genomically integrated pncB gene with an upstream T7,promoter for biotransformation did not result in reproducible increased specific cell productivity. [source] Purification and cDNA Cloning of Lysozyme II from Cabbage Butterfly, Artogeia rapae LarvaeENTOMOLOGICAL RESEARCH, Issue 4 2005BANG In Seok ABSTRACT Last instar larvae of cabbage butterfly Artogeia rapae respond to injection of bacteria with a set of inducible antibacterial peptides/proteins. The inducible peptides/proteins are related to the known hinnavins (I and II) and lysozymes (I and II). The lysozyme II has been isolated by heat treatment, cation exchange, and reversed-phase chromatography from immunized hemolymph of last instar larvae. The lysozyme II gene of A. rapae was isolated and its nucleotide sequence was determined by the RACE-PCR from immunized fat body with E. coli. It has an open reading frame of 414 bp nucleotide corresponding to 138 amino acids including an 18 amino acid signal sequence. The molecular weight and the isoelectric point of Artogeia lysozyme II without a signal peptide were 13,649.38 Da and 9.11, respectively. It is great similarity with Manduca lysozyme among other lepidopteran. [source] Novel DNA repair alkyltransferase from Caenorhabditis elegansENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 2-3 2001Sreenivas Kanugula Abstract O6 -Alkylguanine DNA-alkyltransferase (AGT) is a widely distributed DNA repair protein that protects living organisms from endogenous and exogenous alkylation damage to DNA at the O6 -position of guanine. The search of the C. elegans genome database for an AGT protein revealed the presence of a protein (cAGT-2) with some similarity to known AGTs in addition to the easily recognized cAGT-1 protein. The predicted protein sequence of cAGT-2 contains the amino acid sequence ,ProCysHisPro, at the presumed active site of the protein, whereas all other known AGTs have ,ProCysHisArg,. A truncated version of the cAGT-2 protein was expressed in E. coli. This purified recombinant protein was able to repair O6 -methylguanine and O4 -methylthymine adducts in DNA in vitro and also reacted with the bulky benzyl adduct in O6 -benzylguanine. This fragment of cAGT-2 (104 amino acids) is the smallest protein possessing AGT activity yet described. The full-length cAGT-2 protein (274 amino acids) totally lacks the N-terminal domain present in all other known AGTs but has a long C-terminal extension that has significant homology to histone 1C. Expression of cAGT-2 in an E. coli strain lacking endogenous AGT activity provided modest but statistically significant resistance to the toxicity of N -methyl- N,-nitro- N -nitrosoguanidine, confirming that cAGT-2 is an alkyltransferase. Environ. Mol. Mutagen. 38:235,243, 2001. © 2001 Wiley-Liss, Inc. [source] Characteristics of mutations generated through digestion with restriction enzyme and ligation in plasmid DNAENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 1 2001Shingo Nakamura Abstract Recently, the use of restriction enzymes has been extended to studies in which rare events such as mutation and mistakes in DNA repair are examined. In these studies, the specificity of restriction enzymes becomes critical. To clarify the nature of the rare unexpected events occurring in the process of cutting of DNA with restriction enzymes then ligating it, we studied the molecular characteristics of unexpected plasmid DNAs that were retrieved as mutants of the plasmid after transfection to E. coli. The plasmid used was pUR288, containing lacZ as a marker of mutation. It was digested with restriction enzymes under the conditions recommended by the supplier of the enzymes and under the presence of DMSO, which is known to induce star activity of the enzymes. Comparisons of mutant frequencies and of nucleotide sequences of the mutants found in the different conditions indicated that nonspecific endonucleolytic activity similar to that found under star activity was present under the recommended conditions and, further, was responsible for the creation of deletion-type mutations. The frequency of these events ranged from 10,5 to 10,3, depending on the kind of restriction enzymes analyzed. Although the levels of the nonspecificity were not high, they should be considered in assays such as mutation and mistakes in DNA repair, where rare events are examined. Environ. Mol. Mutagen. 38:46,54, 2001 © 2001 Wiley-Liss, Inc. [source] Ion transport and osmotic adjustment in Escherichia coli in response to ionic and non-ionic osmoticaENVIRONMENTAL MICROBIOLOGY, Issue 1 2009Lana Shabala Summary Bacteria respond to osmotic stress by a substantial increase in the intracellular osmolality, adjusting their cell turgor for altered growth conditions. Using Escherichia coli as a model organism we demonstrate here that bacterial responses to hyperosmotic stress specifically depend on the nature of osmoticum used. We show that increasing acute hyperosmotic NaCl stress above ,1.0 Os kg,1 causes a dose-dependent K+ leak from the cell, resulting in a substantial decrease in cytosolic K+ content and a concurrent accumulation of Na+ in the cell. At the same time, isotonic sucrose or mannitol treatment (non-ionic osmotica) results in a gradual increase of the net K+ uptake. Ion flux data are consistent with growth experiments showing that bacterial growth is impaired by NaCl at the concentration resulting in a switch from net K+ uptake to efflux. Microarray experiments reveal that about 40% of upregulated genes shared no similarity in their responses to NaCl and sucrose treatment, further suggesting specificity of osmotic adjustment in E. coli to ionic and non-ionic osmotica. The observed differences are explained by the specificity of the stress-induced changes in the membrane potential of bacterial cells highlighting the importance of voltage-gated K+ transporters for bacterial adaptation to hyperosmotic stress. [source] Assigning Escherichia coli strains to phylogenetic groups: multi-locus sequence typing versus the PCR triplex methodENVIRONMENTAL MICROBIOLOGY, Issue 10 2008David M. Gordon Summary It is well recognized that Escherichia coli consists of a number of distinct phylo-groups and that strains of the different phylo-groups vary in their ecological niches, life-history characteristics and propensity to cause disease. Consequently, much can be learnt by assigning a strain of E. coli to one of the recognized phylo-groups. A triplex PCR-based method that enables strains of E. coli to be assigned to a phylo-group using a dichotomous key approach based on the presence or absence of two genes (chuA and yjaA) and an anonymous DNA fragment (TSPE4.C2) has been developed. However, the accuracy with which this method assigns strains to their correct phylo-group has not been adequately evaluated. Consequently, 662 strains of E. coli were characterized using a multi-locus sequence typing approach. Unsupervised population assignment algorithms were used to assign strains to phylo-groups based on the multi-locus sequence typing data. The analyses revealed that 85,90% of E. coli strains can be assigned to a phylo-group and that 80,85% of the phylo-group memberships assigned using the Clermont method are correct. However, the accuracy with which strains are assigned to the correct phylo-group depends on their Clermont genotype. For example, strains yielding a Clermont genotype consistent with phylo-groups B1 and B2 are assigned correctly 95% of the time. Strains failing to yield any PCR products using the Clermont method are seldom members of phylo-group A and strains with such a genotype should not be assigned to a phylo-group. [source] A metagenomic analysis of soil bacteria extends the diversity of quorum-quenching lactonasesENVIRONMENTAL MICROBIOLOGY, Issue 3 2008Kashif Riaz Summary A metagenomic library of 10 121 clones, generated from bacteria inhabiting a pasture soil from France, was screened for the presence of fosmids conferring either N -acylhomoserine lactone (NAHL) synthesis or NAHL degradation ability upon their Escherichia coli host. No clone producing NAHLs was identified whereas one, containing a 31 972 bp insert in fosmid p2H8, allowed NAHL degradation. This led to the cloning and identification of a gene, qlcA, encoding an NAHL-lactonase activity, as judged by lactone-ring closure and HPLC/MS analyses of NAHL degradation products. The qlcA gene efficiently quenched quorum-sensing regulated pathogenic functions when expressed in Pectobacterium carotovorum. The QlcA peptide belongs to the family of zinc-dependent metallohydrolases and appears to be distantly related to other NAHL-lactonases discovered in Agrobacterium, Bacillus, Photorhabdus and Rhizobium. In-silico analysis of the metagenomic insert revealed the occurrence of 20 orf, with a constant GC% and codon usage, suggesting a unique bacterial origin. Nine out of these 20 orf were homologous to genes encoding biosynthesis of arginine; they were clustered with an unusual succession argFJADBCRGH. The fosmid p2H8 is able to complement the argA, argB and argC mutants in E. coli. Phylogenetic analysis showed that 9 orf out of 20 were related to sequences from members of the Acidobacteria, supporting the hypothesis that the analysed insert might be originated from an organism related to this phylum. [source] Phenotypic and genotypic characterization of encapsulated Escherichia coli isolated from blooms in two Australian lakesENVIRONMENTAL MICROBIOLOGY, Issue 5 2005Michelle L. Power Summary Escherichia coli has long been used as an indicator organism for water quality assessment. Recently there has been an accumulation of evidence that suggests some strains of this organism are able to proliferate in the environment, a characteristic that would detract from its utility as an indicator of faecal pollution. Phenotypic and genotypic characterization of E. coli isolated from blooms in two Australian lakes, separated by a distance of approximately 200 km, identified that the blooms were dominated by three E. coli strains. A major phenotypic similarity among the three bloom strains was the presence of a group 1 capsule. Genetic characterization of a conserved region of the cps gene cluster, which encodes group 1 capsules, identified a high degree of genetic variation within the bloom isolates. This differs from previously described encapsulated E. coli strains which are highly conserved at the cps locus. The phenotypic or genotypic profiles of the bloom strains were not identified in 435 E. coli strains isolated from vertebrates. The occurrence of these encapsulated strains suggests that some E. coli have evolved a free-living lifestyle and do not require a host in order to proliferate. The presence of the same three strains in bloom events in different geographical regions of a temperate climate, and at different times, indicates that free-living E. coli strains are able to persist in these water reservoirs. This study provides further evidence of circumstances where caution is required in using E. coli as an indicator organism for water quality. [source] Population genetics of Escherichia coli in a natural population of native Australian ratsENVIRONMENTAL MICROBIOLOGY, Issue 6 2000Gulietta M. Pupo Escherichia coli, a normal inhabitant of the intestinal tract of mammals and birds, is a diverse species. Most studies on E. coli populations involve organisms from humans or human-associated animals. In this study, we undertook a survey of E. coli from native Australian mammals, predominantly Rattus tunneyi, living in a relatively pristine environment in the Bundjalung National Park. The genetic diversity was assessed and compared by multilocus enzyme electrophoresis (MLEE), sequence analysis of the mdh (malate dehydrogenase) gene and biotyping using seven sugars. Ninety-nine electrophoretic types were identified from the 242 isolates analysed by MLEE and 15 sequences from the mdh genes sequenced from 21 representative strains. The Bundjalung isolates extend the diversity represented by the E. coli reference (ECOR) set, with new MLEE alleles found in six out of 10 loci. Many of the Bundjalung isolates fell into a discrete group in MLEE. Other Bundjalung strains fell into the recognized E. coli ECOR set groups, but tended to be at the base of both the MLEE and mdh gene trees, implying that these strains are derived independently from ancestral forms of the ECOR groups and that ECOR strains represent only a subset of E. coli adapted to humans and human-associated animals. Linkage disequilibrium analysis showed that the Bundjalung population has an ,epidemic' population structure. The Bundjalung isolates were able to utilize more sugars than the ECOR strains, suggesting that diet plays a prominent role in adaptation of E. coli. [source] Efficiency of permeable pavement systems for the removal of urban runoff pollutants under varying environmental conditionsENVIRONMENTAL PROGRESS & SUSTAINABLE ENERGY, Issue 3 2010Kiran Tota-Maharaj Abstract Urban surface water runoff typically contains a high but variable number of pathogens, nutrients, and sediments that require removal before reuse. Permeable pavements can improve the water quality through interception, filtration, sedimentation, nutrient transformation, and microbial removal. There is currently insufficient scientific information available on the treatment efficiencies of permeable pavements combined with earth energy systems with regards to the removal of storm water pollutants such as nutrients, sediments, and microbial pollutants. This study evaluates the efficiency of 12 tanked combined systems during a medium-term study. The research assessed weekly the removal of the microbial indicators total coliforms, Escherichia coli, and fecal Streptococci, as well as the key nutrients ammonia-nitrogen, nitrate-nitrogen, and ortho-phosphate-phosphorus, and physical variables such as suspended solids and turbidity. Total coliforms, E. coli, and fecal Streptococci were removed by 98,99%. The ammonia-nitrogen and ortho-phosphate-phosphorus removal efficiencies were 84.6% and 77.5%, respectively. An analysis of variance indicated that the presence or absence of a geotextile did result in a very highly statistically significant difference (P < 0.001) with respect to the removal of both ammonia-nitrogen and ortho-phosphate-phosphorus. Suspended solids, turbidity, and biochemical oxygen demand were reduced by 91%, 82%, and 88%, respectively. These results indicate the potential of the proposed novel system in urban runoff pollutant removal and subsequent reuse of the treated water. © 2010 American Institute of Chemical Engineers Environ Prog, 2010 [source] Biological measurement of estrogenic activity in urine and bile conjugates with the in vitro ER-CALUX reporter gene assayENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2002Juliette Legler Abstract Although estrogens are excreted as biologically inactive conjugates, they can be reconverted to an active form, possibly by bacteria. A simple method was developed to deconjugate estrogen metabolites present in human urine and fish bile back to active estrogens by enzymatic hydrolysis with ,-glucuronidase or live Escherichia coli cells. Deconjugated extracts were tested for estrogenic activity in the in vitro stable estrogen receptor,mediated chemical-activated luciferase gene expression (ER-CALUX) assay. Estrogen glucuronides in urine obtained from human males and females were effectively converted to active forms after incubation with ,-glucuronidase or E. coli. The highest estrogenic activity was found in deconjugated metabolites from urine of a pregnant woman, in which levels up to 3,000 nmol estradiol equivalents per liter of urine were found after overnight incubation of urine with E. coli. Bile sampled from male bream and flounder from various freshwater and marine locations was also deconjugated and a good correlation was found between high biliary estrogenic activity and elevated levels of xenoestrogenic activity in surface water as well as in plasma vitellogenin. Therefore, the measurement of deconjugated bile could form a useful (indirect) biomarker for internal dose of xenoestrogens in male fish. [source] Acute toxicity of (chloro-)catechols and (chloro-)catechol-copper combinations in Escherichia coli corresponds to their membrane toxicity in vitroENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 2 2001Nina Schweigert Abstract (Chloro-)catechols are toxic for bacteria and higher organisms, but the mode of action is not yet clearly understood. We have compared the acute toxicity of different chlorinated catechols to Escherichia coli with membrane toxic effects, namely narcosis and uncoupling that we have determined in an in vitro assay. In vitro membrane toxicity was quantified by measuring the accelerated decay of the membrane potential of chromatophores isolated from Rhodobacter sphaeroides. Both acute and membrane toxicity increased with increasing degree of chlorination. Analysis of dose-response curves, pH dependence, and estimated membrane concentrations gave a consistent picture of the mechanisms of membrane toxicity: At pH 7, the higher-chlorinated catechols acted as uncouplers of oxidative and photophosphorylation, and the lower-chlorinated catechols and catechol acted as narcotics. In the case of 3,5-dichlorocatechol and 4-monochlorocatechol at pH 8.8, both mechanisms appeared to contribute to the overall toxicity. Copper exhibited a diverging effect on the toxicity of catechols and of (chloro-)catechols to E. coli. Whereas the presence of copper increased the toxicity of catechol and 4-monochlorocatechol, the toxicity of 3,5-dichlorocatechol, 3,4,5-trichlorocatechol, and tetrachlorocatechol decreased. Again, the results obtained with in vitro assays agreed with the acute toxicity observed in E. coli: The presence of copper accelerated decay of the membrane potential of catechol and 4-monochlorocatechol; however, the effect was reversed by copper in experiments with 3,5-dichlorocatechol, 3,4,5-trichlorocatechol, and tetrachlorocatechol. We have proposed a mechanistic model to explain the diverging effects of copper on the uncoupling activities of the different catechols. [source] A survey of equine abortion, stillbirth and neonatal death in the UK from 1988 to 1997EQUINE VETERINARY JOURNAL, Issue 5 2003K. C. SMITH Summary Reasons for performing study: A detailed review of laboratory records for equine abortion is fundamental in establishing current disease trends and suggesting problems important for further research. Objectives: To review the causes of abortion and neonatal death in equine diagnostic submissions to the Animal Health Trust over a 10 year period. Methods: The diagnoses in 1252 equine fetuses and neonatal foals were reviewed and analysed into categories. Results: Problems associated with the umbilical cord, comprising umbilical cord torsion and the long cord/cervical pole ischaemia disorder, were the most common diagnoses (38.8%: 35.7% umbilical cord torsion and 3.1% long cord/cervical pole ischaemia disorder). Other noninfective causes of abortion or neonatal death included twinning (6.0%), intrapartum stillbirth (13.7%) and placentitis, associated with infection (9.8%). E. coli and Streptococcus zooepidemicus were the most common bacteria isolated. Neonatal infections not associated with placentitis accounted for 3.2% of incidents; and infections with EHV-1 or EHV-4 for 6.5%. Conclusions: Definitive diagnosis of equine abortion is possible in the majority of cases where the whole fetus and placenta are submitted for examination. Potential relevance: Given the high incidence of umbilical cord torsion and related problems as causes of abortion in UK broodmares, more research on factors determining umbilical cord length and risk of torsion is essential. [source] Retinol binding protein isolated from acute renal failure patients inhibits polymorphonuclear leucocyte functionsEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 11 2004G. Cohen Abstract Background, Protein factors accumulating in sera of patients with end-stage renal disease (ESRD) that interfere with the nonspecific immune response by inhibiting essential functions of polymorphonuclear leucocytes (PMNLs) have previously been described. No such factor has been isolated from acute renal failure (ARF) patients to date. Materials and methods, Using a three-step chromatographic procedure involving ion exchange, size exclusion and hydrophobic interaction chromatography we purified the apo- and holo-form of retinol binding protein (RBP) from high-flux dialyser (polyacrylonitrile; AN69) ultrafiltrates of patients with ARF. Their effect on the chemotaxis of PMNLs isolated from healthy donors was determined by the under-agarose method. Whole-blood assays applying flow cytometry were used to assess phagocytosis and the oxidative metabolism of PMNLs. Apoptosis was assessed by determining the DNA content using propidium iodide. Results, Isolated apo- and holo-forms of RBP were truncated on their C-terminus as determined by mass spectrometry. All isolates significantly inhibited the chemotactic movement of PMNLs obtained from healthy donors and the PMNL oxidative metabolism stimulated by E. coli. These effects were concentration dependent. Retinol binding protein had no influence on the PMNL oxidative metabolism stimulated by PMA and on PMNL phagocytosis. Commercially available RBP isolated from urine influenced PMNL functions in the same way. Inhibition of p38 mitogen-activated protein kinase (MAPK) by SB203580 significantly attenuated the phagocytosis-induced respiratory burst and RBP did not lead to a further decrease. Polymorphonuclear leucocyte apoptosis was significantly inhibited by RBP. Conclusions, The apo- and holo-forms of RBP isolated from the ultrafiltrate of ARF patients inhibit PMNL chemotaxis, oxidative metabolism and apoptosis. Therefore, RBP may be considered a uraemic toxin contributing to a disturbed immune defence. [source] Comparative study on antibodies to human and bacterial 60 kDa heat shock proteins in a large cohort of patients with coronary heart disease and healthy subjectsEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 4 2001Z. Prohászka Background Recent observations indicate an association between antibodies against mycobacterial heat shock protein (hsp65) and coronary heart disease (CHD). Previously, we reported on marked differences in antigen specificity and complement activating ability of anti-hsp65 antibodies and auto-antibodies against human heat shock protein, hsp60. Here, we investigated whether there are differences between antih-sp65 and anti-hsp60 antibodies in their association with CHD. Design We measured by ELISA the levels of antibodies to hsp65, hsp60 and E. coli -derived GroEL in three groups: Group I, 357 patients with severe CHD who underwent by-pass surgery; Group II, 67 patients with negative coronary angiography; Group III, 321 healthy blood donors. Antibodies against Helicobacter pylori were also measured by commercial ELISA. Results As calculated by multiple regression analysis, the levels of anti-hsp60 auto-antibodies were significantly higher in Group I compared to Group II (P = 0·007) or Group III (P < 0·0001). By contrast, although concentrations of anti-hsp65 and anti-GroEL antibodies in Group I were higher than in Group III, no significant differences between Group I and Group II were found. Antibodies to the two bacterial hsp strongly correlated to each other, but either did not correlate or weakly correlated to hsp60. In Group I, serum concentrations of anti- H.pylori antibodies significantly correlated with those of anti-hsp65 and anti-GroEL antibodies but they did not correlate with the anti-hsp60 antibodies. Conclusion As to their clinical relevance, a remarkable difference become evident between antibodies to human hsp60 and antibodies against bacterial hsp in the extent of association with CHD. On the basis of these findings and some pertinent literature data, an alternative explanation for the association between high level of anti-hsp antibodies and atherosclerotic vascular diseases is raised. [source] Antibiotic-Loaded PLGA Nanofibers for Wound Healing Applications,ADVANCED ENGINEERING MATERIALS, Issue 4 2010David A. Soscia Incorporating antibiotics into biocompatible nanoscale non-woven fibrous mats could provide utility for wound healing applications and for incorporation into wound dressing materials. In this study, the antibiotic chloramphenicol (Cm) was incorporated into electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers, which were then tested for inhibition of bacterial growth for multiple bacterial species (Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella typhimurium, and Pseudomonas aeruginosa). In addition, the cytotoxicity of Cm-PLGA nanofibers was examined for two types of mammalian cells including mouse embryonic stem cells and fibroblasts. Electrospun PLGA nanofibers containing Cm were able to reduce bacterial growth on solid agar plates for all species except for P. aeruginosa. In liquid culture, Cm-loaded nanofibers inhibited growth for E. coli, B. cereus and S. typhimurium by 93% or greater, while P. aeruginosa and S. aureus growth was inhibited by 42% and 56%, respectively. Cm-loaded nanofibers showed limited cytoxicity on fibroblasts and embryonic stem cells, with viability greater than 96% for all conditions tested. These results suggest that Cm can be successfully incorporated into electrospun nanofibers and that these fibers could be used for wound healing applications with minimal cytotoxicity to the surrounding tissue. [source] Synthesis of Cluster Mannosides Carrying a Photolabile Diazirine GroupEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 3 2006Mark Walter Abstract We are investigating the mechanisms underlying the carbohydrate-specific adhesion of bacteria such as Escherichia coli to the glycocalyx of their potential host cells. E. coli possess protein appendages, which are called type 1 fimbriae. Part of type 1 fimbriae is a protein named FimH, which is a mannose-specific lectin. We wish to use photoaffinity labeling to elucidate mannose binding sites on FimH. Thus we report the synthesis of di- and trivalent cluster mannosides, which carry a photolabile diazirine group. The diazirine group was introduced by a convergent approach using thiourea bridging (products 6, 13, 17, and 27) or in a divergent synthesis leading to the divalent cluster mannoside 31. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006) [source] Glycerol and Glycerol Glycol GlycodendrimersEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 22 2003Mike M. K. Boysen Abstract Non-covalent interactions between structural parts of complex oligosaccharides and saccharide-recognising proteins are of crucial importance for many cell communication phenomena. Specificity of such interactions and stability of these ligand-receptor complexes are achieved through multivalent interactions between multiple copies of a saccharide ligand and a corresponding number of protein receptors. Substances presenting multiple copies of the saccharide ligand on easily accessible scaffold molecules therefore appear to be promising tools for study of multivalent interactions and their possible inhibition. Such multivalent glycomimetics can be prepared by attachment of saccharide residues to the surface functional groups of dendrimers. In the course of our work, we have prepared novel glycodendrimers with glycerol and glycerol glycol polyether scaffolds. Isopropylidene-protected hydroxyethyl mannoside was chosen as the carbohydrate component, with the construction of the dendritic structures proceeding by a convergent approach featuring iterative Williamson etherification and ozonolysis/hydride reduction steps. Deprotected representatives of such structures are potential inhibitors of mannose-binding lectins of E. coli. Three representative compounds were deprotected and their anti-adhesive properties were examined. The route to these glycodendrimers was also evaluated in terms of synthetic chemistry. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003) [source] Supernatants of Pseudomonas aeruginosa induce the Pseudomonas -specific antibiotic elafin in human keratinocytesEXPERIMENTAL DERMATOLOGY, Issue 4 2003Ulf Meyer-Hoffert Abstract: Elafin is a skin-derived serine-protease inhibitor. It is thought to be important to prevent human leukocyte elastase-mediated tissue damage and might play an important role in maintaining the integrity of the human epidermis. Recent studies have provided evidence for an antimicrobial activity of elafin against P. aeruginosa. As gram-negative infections typically occur in barrier-disrupted skin we were interested to determine whether supernatants of the gram-negative bacteria P. aeruginosa and Escherichia coli were capable of inducing elafin expression. Supernatants of various P. aeruginosa strains stimulated elafin mRNA-expression and protein release, whereas supernatants of E. coli did not induce elafin expression. In non-differentiated cells the relative increase of elafin mRNA was much higher (100-fold) than in differentiated cells (sixfold), although the latter exhibited higher constitutive mRNA-expression (150-fold). However, concentrations of secreted elafin were similar in differentiated and non-differentiated cells after stimulation. We could not confirm a bactericidal effect against P. aeruginosa as described previously but observed that its growth was inhibited as demonstrated for different strains in liquid cultures. Growth of E. coli was not affected by elafin. In conclusion, the data presented in this paper suggest that elafin represents an innate immune response factor induced by secreted products of P. aeruginosa. Besides its elastase inhibitory potency elafin is an antimicrobial agent against P. aeruginosa. [source] Adrenomedullin and Proadrenomudullin N-Terminal 20 Peptide (PAMP) are Present in Human Colonic Epithelia and Exert an Antimicrobial EffectEXPERIMENTAL PHYSIOLOGY, Issue 5 2001K. Marutsuka The hypotensive and vasorelaxing peptides adrenomedullin (AM) and its gene-related peptide, proadrenomedullin N-terminal 20 peptide (PAMP), were found to be distributed on the surface of the colonic mucosa. AM and PAMP showed dose-dependent antimicrobial activity against E. coli. The results suggest that the novel vasoactive peptides AM and PAMP play an important role in mucosal defence. [source] Contact-Killing Polyelectrolyte Microcapsules Based on Chitosan DerivativesADVANCED FUNCTIONAL MATERIALS, Issue 19 2010Di Cui Abstract Polyelectrolyte-multilayer microcapsules are made by layer-by-layer (LbL) assembly of oppositely charged polyelectrolytes onto sacrificial colloidal particles, followed by core removal. In this paper, contact-killing polyelectrolyte microcapsules are prepared based solely on polysaccharides. To this end, water-soluble quaternized chitosan (QCHI) with varying degrees of substitution (DS) and hyaluronic acid (HA) are assembled into thin films. The quaternary ammonium groups are selectively grafted on the primary amine group of chitosan by exploiting its reaction with glycidyltrimethylammonium chloride (GTMAC) under homogeneous aqueous acidic conditions. The morphology of the capsules is closely dependent on the DS of the quaternized chitosan derivatives, which suggests differences in their complexation with HA. The DS is also a key parameter to control the antibacterial activity of QCHI against Escherichia Coli (E. coli). Thus, capsules containing the QCHI derivative with the highest DS are shown to be the most efficient to kill E. coli while retaining their biocompatibility toward myoblast cells, which suggests their potential as drug carriers able to combat bacterial infections. [source] | |||||||||||||||||||||||||||||||||||||