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a plastic microchip for nucleic acid purification.a microchip for purifying nucleic acids from bacterial pathogens was designed and fabricated in plastic. the fabricated plastic microchips were tested for their ability to purify nucleic acids from the bacteria listeria monocytogenes (l. monocytogenes), escherichia coli (e. coli), and salmonella typhimurium (s. typhimurium). these chips were constructed using rapid and low-cost plastic fabrication techniques including hot embossing and plastic casting. silicon molds fabricated by photolithograph ...200717530410
a rapid multiplexed chemiluminescent immunoassay for the detection of escherichia coli o157:h7, yersinia enterocolitica, salmonella typhimurium, and listeria monocytogenes pathogen bacteria.a simple and rapid multiplexed sandwich chemiluminescent enzyme immunoassay has been developed for the simultaneous detection of escherichia coli o157:h7, yersinia enterocolitica, salmonella typhimurium, and listeria monocytogenes. to achieve the multiplexed detection of the four pathogens, a new polystyrene 96 well microtiter plate format has been designed, in which each main well contains four subwells in the bottom. the monoclonal antibodies specific for each bacteria were separately immobili ...200717530767
characterization of a broad range antibacterial substance from a new bacillus species isolated from amazon basin.a bacillus sp. strain producing a bacteriocin-like substance was characterized by biochemical profiling and 16s rdna sequencing. the phylogenetic analysis indicated that this strain has low sequence similarity with most bacillus spp., suggesting a new species was isolated. the antimicrobial activity was detected starting at the exponential growth phase, and maximum activity was observed at stationary phase. the substance was inhibitory to a broad range of indicator strains, including pathogenic ...200717534601
application of automated ribotyping to support the evaluation of listeria monocytogenes sources in a taleggio cheese producing plant.in march 2005, listeria monocytogenes was detected on the rinds of taleggio cheeses produced in an italian plant. to identify the pathogen source, 154 rinds of cheeses that had been manually and automatically salinated and 52 environmental swabs collected from salting equipment, ripening cloths, and ripening boxes were tested for l. monocytogenes. twenty-seven strains isolated from cheese samples and 16 strains isolated from environmental samples were genotyped by ecori and pvuii automated ribot ...200717536669
dose-response of listeria monocytogenes after oral exposure in pregnant guinea pigs.listeriosis, a severe disease that results from exposure to the foodborne pathogen listeria monocytogenes, is responsible for approximately 2500 illnesses and 500 deaths in the united states each year. pregnant women are 20 times more likely to develop listeriosis than the general population, with adverse pregnancy outcomes that include spontaneous abortions, stillbirths, and neonatal meningitis. the objective of this study was to determine an infective dose that resulted in stillbirths and infe ...200717536670
proteomic analysis of a hypochlorous acid-tolerant listeria monocytogenes cultural variant exhibiting enhanced biofilm production.following exposure of listeria monocytogenes scott a (sa) to hypochlorous acid, rough colony variants were identified that were tolerant of hypochlorous acid and produced increased amounts of biofilm. a derivative of one of these variants was smooth, produced even more biofilm, and exhibited greater biofilm chlorine resistance. the objective of this research was to compare the protein expression of a cultural variant to sa and to identify proteins that might be associated with biofilm production ...200717536671
extracting additional risk managers information from a risk assessment of listeria monocytogenes in deli meats.the risk assessment study of listeria monocytogenes in ready-to-eat foods conducted by the u.s. food and drug administration is an example of an extensive quantitative microbiological risk assessment that could be used by risk analysts and other scientists to obtain information and by managers and stakeholders to make decisions on food safety management. the present study was conducted to investigate how detailed sensitivity analysis can be used by assessors to extract more information on risk f ...200717536672
antimicrobial activity of olive oil, vinegar, and various beverages against foodborne pathogens.the survival of foodborne pathogens in aqueous extracts of olive oil, virgin olive oil, vinegar, and several beverages was evaluated. vinegar and aqueous extracts of virgin olive oil showed the strongest bactericidal activity against all strains tested. red and white wines also killed most strains after 5 min of contact, black and green tea extracts showed weak antimicrobial activity under these conditions, and no effect was observed for the remaining beverages (fruit juices, coca-cola, dairy pr ...200717536679
inhibitory activity of colicin e1 against listeria monocytogenes.colicins are gram-negative bacteriocins produced by and effective against escherichia coli and related species. colicin e1 (cole1) is composed of three functional domains, which collectively have a pore-forming effect on targeted bacteria. cole1 binding and translocation domains are highly specific in contrast to the pore-forming domain, implying that cole1 could be broadly effective. in this study, the activity of cole1 against listeria monocytogenes was evaluated in broth and on surfaces of re ...200717536690
use of one-ply composite tissues in an automated optical assay for recovery of listeria from food contact surfaces and poultry-processing environments.a novel one-ply composite tissue (ct) method using the soleris (formerly biosys) optical analysis system was compared with the conventional u.s. department of agriculture (usda) environmental sponge enrichment method for recovery of listeria from food contact surfaces and poultry-processing environments. stainless steel and high-density polyethylene plates were inoculated to contain a six-strain l. monocytogenes cocktail at 10(4), 10(2), and 10 cfu per plate, whereas samples from naturally conta ...200717536691
effectiveness and stability of plastic films coated with nisin for inhibition of listeria monocytogenes.plastic films were coated with a cellulose-based carrier solution containing nisin, a natural antimicrobial peptide with the potential to inhibit growth of food spoilage and pathogenic microorganisms such as listeria monocytogenes. five commercial plastic films with different chemical compositions and surface properties were compared in this study: low-density polyethylene, ethylene-vinyl acetate copolymer, and three types of ethylene-methacrylic acid copolymers: surlyn 1601, nucrel 0403, and nu ...200717536692
minimally processed vegetable salads: microbial quality evaluation.the increasing demand for fresh fruits and vegetables and for convenience foods is causing an expansion of the market share for minimally processed vegetables. among the more common pathogenic microorganisms that can be transmitted to humans by these products are listeria monocytogenes, escherichia coli o157:h7, and salmonella. the aim of this study was to evaluate the microbial quality of a selection of minimally processed vegetables. a total of 181 samples of minimally processed leafy salads w ...200717536694
an overview of microbial food safety programs in beef, pork, and poultry from farm to processing in canada.canada's vision for the agri-food industry in the 21st century is the establishment of a national food safety system employing hazard analysis and critical control point (haccp) principles and microbiological verification tools, with traceability throughout the gate-to-plate continuum. voluntary on-farm food safety (offs) programs, based in part on haccp principles, provide producers with guidelines for good production practices focused on general hygiene and biosecurity. offs programs in beef c ...200717536696
impact of selected lactobacillus and bifidobacterium species on listeria monocytogenes infection and the mucosal immune response.the pathogenesis of listeria monocytogenes depends on its ability to attach to and invade the gastrointestinal epithelium and subsequently withstand the host immune response. despite a thorough understanding of the intracellular phase of infection, relatively little is known about how the pathogen behaves in the gastrointestinal tract and whether it is affected by the presence of host commensal microbiota. lactobacillus and bifidobacterium are two important genera of the human gut microbiota pro ...200717537177
a predictive model for the influence of food components on survival of listeria monocytogenes lm 54004 under high hydrostatic pressure and mild heat conditions.the combination of high hydrostatic pressure with mild temperature was explored to achieve a predictive model of microbial inactivation in food matrix processing. the pressure processing conditions were fixed at 448 mpa for 11 min at the treatment temperature of 41 degrees c, which have been determined as the optimum processing conditions considering six log-cycle reductions of listeria monocytogenes. based on the results, response surface methodology (rsm) was performed in the present work, the ...200717537535
avirulent viable but non culturable cells of listeria monocytogenes need the presence of an embryo to be recovered in egg yolk and regain virulence after recovery.the aim of this study was to assess the efficiency of the embryonated egg model to recover viable but non culturable (vbnc) cells of listeria monocytogenes. l. monocytogenes cells were incubated in filtered sterilised distilled water. the vbnc state was obtained after a 25 to 47 days incubation period (concentration of culturable cells less than 1 cfu/ml). fifteen days after the vbnc state was reached, non culturability was checked in various media. one milliliter of each vbnc suspension that co ...200717540159
extending the host range of listeria monocytogenes by rational protein design.in causing disease, pathogens outmaneuver host defenses through a dedicated arsenal of virulence determinants that specifically bind or modify individual host molecules. this dedication limits the intruder to a defined range of hosts. newly emerging diseases mostly involve existing pathogens whose arsenal has been altered to allow them to infect previously inaccessible hosts. we have emulated this chance occurrence by extending the host range accessible to the human pathogen listeria monocytogen ...200717540170
synthesis, characterization and antimicrobial activity of new aliphatic sulfonamide.a series of novel aliphatic sulfonamide derivatives (1-7) were synthesized and characterized by elemental analyses, ft-ir, (1)h nmr, (13)c nmr and lc-ms techniques. all the synthesized compounds were evaluated in vitro as antimicrobial agents against representative strains of gram-positive (staphylococcus aureus atcc 25953, bacillus cereus atcc 6633 and listeria monocytogenes atcc li6 (isolate), gram-negative bacteria (escherichia coli atcc 11230) and antifungal agent against candida albicans (c ...200717544281
rates of processing determine the immunogenicity of immunoproteasome-generated epitopes.cd8 t cells resolve intracellular pathogens by responding to pathogen-derived peptides that are presented on the cell surface by mhc class i molecules. although most pathogens encode a large variety of antigenic peptides, protective cd8 t cell responses target usually only a few of these. to determine the mechanism by which the ifn-gamma-inducible proteasome (immuno) subunits enhance the ability of specific pathogen-derived peptides to elicit cd8 t cell responses, we generated a recombinant list ...200717548590
allelic penetrance approach as a tool to model two-locus interaction in complex binary traits.many binary phenotypes do not follow a classical mendelian inheritance pattern. interaction between genetic and environmental factors is thought to contribute to the incomplete penetrance phenomena often observed in these complex binary traits. several two-locus models for penetrance have been proposed to aid the genetic dissection of binary traits. such models assume linear genetic effects of both loci in different mathematical scales of penetrance, resembling the analytical framework of quanti ...200717551528
[epidemiology of listeriosis in germany: a changing but ignored pattern].listeriosis is emerging as a food-borne disease. although the incidence of clinical presentations is low, this infectious disease with a rather poor prognosis deserves attention. approximately 20-30% of patients with recognized listeriosis die in spite of an appropriate antibiotic regimen. in the last two years the number of registered cases of listeriosis in germany has more than doubled. this increase is clearly due exclusively to the rising number of infections in elderly persons, while the n ...200717551889
[mode of action of plantaricin l-1, an antilisteria bacteriocin produced by lactobacillus plantarum].plantaricin l-1, an anti-listeria bacteriocin, was produced by lactobacillus plantarum and successfully purified by sp-sepharose ff cation exchange chromatography. the mechanism on energized cells of listeria monocytogenes was studied with purified plantaricin l-1. after adding plantaricin l-1 to listeria monocytogenes at 64 au/ml, leakage of intercellular k+ ions, inorganic phosphate, lactic dehydrogenase, uv-absorbing materials and the intracellular atp was observed, and the action resulted in ...200717552231
culture-positive sepsis in neonatal camelids: 21 cases.there is limited literature on neonatal bacterial sepsis in new world (nw) camelids.200717552461
listeria monocytogenes surface proteins: from genome predictions to function.the genome of the human food-borne pathogen listeria monocytogenes is predicted to encode a high number of surface proteins. this abundance likely reflects the ability of this bacterium to survive in diverse environments, including soil, food, and the human host. this review focuses on the various mechanisms by which listerial proteins are attached at the bacterial surface and their many functions, including peptidoglycan metabolism, protein processing, adhesion to host cells, and invasion of ho ...200717554049
type ii phosphatidylinositol 4-kinases promote listeria monocytogenes entry into target cells.interaction of the listeria surface protein inlb with the hepatocyte growth factor receptor met activates signalling events that trigger bacterial internalization into mammalian epithelial cells. we show here that purified phagosomes containing inlb-coated beads display type ii phosphatidylinositol 4-kinase (pi4k) activity. in human epithelial hela cells, both pi4kiialpha and pi4kiibeta isoforms are corecruited with met around inlb-coated beads or wild-type listeria during the early steps of int ...200717555516
initial t cell receptor transgenic cell precursor frequency dictates critical aspects of the cd8(+) t cell response to infection.adoptive-transfer experiments with relatively large input numbers ( approximately 10(6)) of t cell receptor-transgenic (tcr-tg) t cells are widely used to model endogenous t cell responses to infection or immunization. we show that input numbers of naive tcr-tg t cells sufficient to squelch the endogenous response to the same epitope substantially alter the kinetics, proliferative expansion, phenotype, and efficiency of memory generation by the tcr-tg t cells in response to infection. thus, resp ...200717555991
probabilistic modeling approach for evaluating the compliance of ready-to-eat foods with new european union safety criteria for listeria monocytogenes.among the new microbiological criteria that have been incorporated in eu regulation 2073/2005, of particular interest are those concerning listeria monocytogenes in ready-to eat (rte) foods, because for certain food categories, they no longer require zero tolerance but rather specify a maximum allowable concentration of 100 cfu/g or ml. this study presents a probabilistic modeling approach for evaluating the compliance of rte sliced meat products with the new safety criteria for l. monocytogenes ...200717557858
nonsense-mutated inla and prfa not widely distributed in listeria monocytogenes isolates from ready-to-eat seafood products in japan.inla is a surface protein participating in the entry of listeria monocytogenes into mammalian non-phagocytic cells. prfa is a positive regulatory factor that regulates the expression of a set of virulence genes. recent studies revealed that some l. monocytogenes strains have a truncated form of these proteins because of nonsense mutations in their sequences, and these truncations contribute to the significant reduction in virulence of this pathogen. in this study, sequence analyses of inla and p ...200717566579
antibacterial properties and major bioactive components of cinnamon stick (cinnamomum burmannii): activity against foodborne pathogenic bacteria.cinnamomum burmannii blume (cinnamon stick) from indonesia is a little-investigated spice. in this study, the antibacterial activity, minimum inhibitory concentration (mic), and minimum bactericidal concentration (mbc) of cinnamon stick extract were evaluated against five common foodborne pathogenic bacteria (bacillus cereus, listeria monocytogenes, staphylococcus aureus, escherichia coli, and salmonella anatum). cinnamon stick extract exhibited significant antibacterial properties. major compou ...200717567030
structure and mechanism of the genomically encoded fosfomycin resistance protein, fosx, from listeria monocytogenes.the fosfomycin resistance protein, fosx, catalyzes the hydration of the antibiotic fosfomycin, (1r,2s)-epoxypropylphosphonic acid. genes encoding the enzyme are found in several pathogenic microorganisms. the structure and mechanism of action of the genomically encoded fosx enzyme from listeria monocytogenes (fosxlmatcc) obtained from the american type culture collection are reported. the gene harbors 31 point mutations, and as a consequence, the protein differs in 10 amino acid residues from th ...200717567049
listeria monocytogenes evades killing by autophagy during colonization of host cells.listeria monocytogenes is an intracellular pathogen that is able to colonize the cytosol of macrophages. here we examined the interaction of this pathogen with autophagy, a host cytosolic degradative pathway that constitutes an important component of innate immunity towards microbial invaders. l. monocytogenes infection induced activation of the autophagy system in macrophages. at 1 h post infection (p.i.), a population of intracellular bacteria ( approximately 37%) colocalized with the autophag ...200717568179
oxygen restriction increases the infective potential of listeria monocytogenes in vitro in caco-2 cells and in vivo in guinea pigs.listeria monocytogenes has been implicated in several food borne outbreaks as well as sporadic cases of disease. increased understanding of the biology of this organism is important in the prevention of food borne listeriosis. the infectivity of listeria monocytogenes scotta, cultivated with and without oxygen restriction, was compared in vitro and in vivo. fluorescent protein labels were applied to allow certain identification of listeria cells from untagged bacteria in in vivo samples, and to ...200717570840
[listeria monocytogenes encephalitis revealed by left progressive hemiparesis].central nervous system listeriosis is a diagnostic and therapeutic challenge for the clinician.200717571028
[listeriosis in a patient with hemodialysis and iron overload].a case of listeriosis in a patient undergoing long-term hemodialysis is reported. the 69-year-old man with chronic renal failure due to polycystic renal dysplasia had received hemodialysis treatment for 10 years. renal anemia and iron overload developed within this time. in the course of a hemodialysis the patient presented with malaise, chills and fever to 38.7 degrees c. blood culture specimens were obtained and the patient received ampicillin-sulbactam. blood culture specimens subsequently sh ...200717571224
inactivation of escherichia coli and listeria monocytogenes on iceberg lettuce by dip wash treatments with organic acids.to study and compare the efficacy of organic acids and chlorine dipping in inactivation of escherichia coli and listeria monocytogenes on fresh-cut iceberg lettuce.200717576223
novel protein targets of the humoral immune response to listeria monocytogenes infection in rabbits.the role of the humoral immune response in protective immunity against listerial infection has been overlooked and is essentially unknown. this study aimed to discover the protein targets of listeria monocytogenes that elicit an antibody response following infection in a rabbit model. a genomic expression library for l. monocytogenes was constructed and differentially screened to identify genes encoding proteins that reacted with antiserum from rabbits infected with live l. monocytogenes serotyp ...200717577052
action of divergicin m35, a class iia bacteriocin, on liposomes and listeria.the mode of action of divergicin m35, a class iia bacteriocin, was studied against listeria monocytogenes with sensitive (divs) and resistant (divm) phenotypes, as well as on synthetic phospholipid liposomes.200717578415
identification of a macrophage antigen-processing event required for i-region-restricted antigen presentation to t lymphocytes. j. immunol. 1981. 127: 1869-1875. 200717579013
manipulating the rate of memory cd8+ t cell generation after acute infection.infection with listeria monocytogenes elicits expansion in numbers of ag-specific cd8+ t cells, which then undergo programmed contraction. the remaining cells undergo further phenotypic and functional changes with time, eventually attaining the qualities of memory cd8+ t cells. in this study, we show that l. monocytogenes-specific cd8+ t cell populations primed in antibiotic-pretreated mice undergo brief effector phase, but rapidly develop phenotypic (cd127(high), cd43(low)) and functional (gran ...200717579021
a reduced antigen load in vivo, rather than weak inflammation, causes a substantial delay in cd8+ t cell priming against mycobacterium bovis (bacillus calmette-guérin).regardless of the dose of ag, ag presentation occurs rapidly within the first few days which results in rapid expansion of the cd8+ t cell response that peaks at day 7. however, we have previously shown that this rapid priming of cd8+ t cells is absent during infection of mice with mycobacterium bovis (bacillus calmette-guérin (bcg)). in this study, we have evaluated the mechanisms responsible for the delayed cd8+ t cell priming. because bcg replicates poorly and survives within phagosomes we co ...200717579040
nod1/rick and tlr signaling regulate chemokine and antimicrobial innate immune responses in mesothelial cells.mesothelial cells that line the serous cavities and outer surface of internal organs are involved in inflammatory responses induced by microbial stimuli and bacterial infection. upon exposure to bacterial products, mesothelial cells secrete chemokines, but the signaling pathways by which these cells recognize bacteria to mediate innate immune responses remain largely unknown. we report that stimulation of primary peritoneal mesothelial cells via nucleotide-binding oligomerization domain (nod)1, ...200717579072
listeria infections associated with infliximab: case reports.infliximab is a human-murine chimeric monoclonal antibody directed against tumor necrosis factor alpha (tnfalpha). infliximab and other tnf blockers are used to treat inflammatory diseases such as rheumatoid arthritis (ra). tnf blockers are suspected to play a key role in some infections. we report here two cases of listeria monocytogenes sepsis associated with infliximab treatment for ra. the first patient developed a terminal ileitis and a bacteraemia after three doses of infliximab; the secon ...200717579802
evaluation of quantitative recovery methods for listeria monocytogenes applied to stainless steel.the ability of listeria monocytogenes to attach to various food contact surfaces, such as stainless steel, polypropylene, and rubber compounds, is well documented. the retention of these or other pathogenic bacteria on food contact surfaces increases the risk of transmission to food products. the objective of this study was to compare several methods for quantitative recovery of listeria monocytogenes from stainless steel surfaces. a cocktail of 4 serotypes of listeria monocytogenes mixed in equ ...200717580634
acid adaptation and survival of listeria monocytogenes in italian-style soft cheeses.the ability of listeria monocytogenes to survive and grow at high salt concentrations and low ph makes it a potential hazard after the consumption of milk and dairy products, often implicated in severe outbreaks of listeriosis. this study was designed to evaluate the behaviour of l. monocytogenes in traditional acid and salted italian-style soft cheeses and to investigate whether listeria occurrence and growth in these environments may represent a potential increase of hazard.200717584464
microplate fluorescence assay for measurement of the ability of strains of listeria monocytogenes from meat and meat-processing plants to adhere to abiotic surfaces.listeria monocytogenes is a significant food-borne pathogen that is capable of adhering to and producing biofilms on processing equipment, making it difficult to eliminate from meat-processing environments and allowing potential contamination of ready-to-eat (rte) products. we devised a fluorescence-based microplate method for screening isolates of l. monocytogenes for the ability to adhere to abiotic surfaces. strains of l. monocytogenes were incubated for 2 days at 30 degrees c in 96-well micr ...200717586676
characterization of new l,d-endopeptidase gene product cwlk (previous ycdd) that hydrolyzes peptidoglycan in bacillus subtilis.bacillus subtilis has various cell wall hydrolases, however, the functions and hydrolase activities of some enzymes are still unknown. b. subtilis cwlk (ycdd) exhibits high sequence similarity with the peptidoglycan hydrolytic l,d-endopeptidase (ply500) of listeria monocytogenes phage and cwlk has the vany motif which is a d-alanyl-d-alanine carboxypeptidase (pfam: http://www.sanger.ac.uk/software/pfam/). the beta-galactosidase activity observed on cwlk-lacz fusion indicated that the cwlk gene w ...200717588176
sepsis and meningitis due to listeria monocytogenes.this study focused on the effect of immuno-compromising conditions on the clinical presentation of severe listerial infection. patients and200717594151
epsilon-poly-l-lysine and nisin a act synergistically against gram-positive food-borne pathogens bacillus cereus and listeria monocytogenes.to test epsilon-poly-l-lysine and nisin a for possible synergistic activity against selected food-borne pathogens and food spoilage organisms.200717594454
irreversible loss of membrane-binding activity of listeria-derived cytolysins in non-acidic conditions: a distinct difference from allied cytolysins produced by other gram-positive bacteria.listeriolysin o (llo), a member of the cholesterol-dependent cytolysin (cdc) family, is a major virulence factor of listeria monocytogenes and contributes to bacterial escape from intracellular killing of macrophages. llo is activated under weakly acidic conditions; however, the molecular mechanism of this ph-dependent expression of cytolytic activity of llo is poorly understood. in this study, cdcs including llo, ivanolysin o (ilo), seeligeriolysin o (lso), pneumolysin (ply), streptolysin o (sl ...200717600069
antimicrobial susceptibility of listeria monocytogenes from food products.this study was conducted to determine the susceptibility of listeria monocytogenes isolated from food products to antimicrobial agents commonly used for treatment of infections with gram-positive bacteria, and to disinfectants. a total of 114 l. monocytogenes retail isolates were tested for susceptibility to ceftiofur, chloramphenicol, ciprofloxacin, erythromycin, florfenicol, penicillin, spectinomycin, streptomycin, tetracycline, tiamulin, trimethoprim, and co-trimoxazole, and the disinfectants ...200717600489
distribution of internalin gene profiles of listeria monocytogenes isolates from different sources associated with phylogenetic lineages.listeria monocytogenes is a human foodborne pathogen with a broad range of hosts. while the l. monocytogenes genome encodes a number of internalins, which are leucine-rich repeat bacterial surface proteins with putative or confirmed roles in host cell attachment and invasion, the specific function of many internalins remains unclear. the distribution of 7 internalin genes (inlc, inlc2, inld, inle, inlf, inlg, and inlh) in 120 l. monocytogenes isolates from humans, foods, and animals was investig ...200717600490
introduction to united states department of agriculture vetnet: status of salmonella and campylobacter databases from 2004 through 2005.in 2003 the united states department of agriculture established usda vetnet. it was modeled after pulsenet usa, the national molecular subtyping network for foodborne disease surveillance. the objectives of usda vetnet are: to use pulsed-field gel electrophoresis (pfge) to subtype zoonotic pathogens submitted to the animal arm of the national antimicrobial resistance monitoring system (narms); examine vetnet and pulsenet pfge patterns; and use the data for surveillance and investigation of suspe ...200717600492
construction and evaluation of a microbiological positive process internal control for pcr-based examination of food samples for listeria monocytogenes and salmonella enterica.pcr assays for food-borne pathogens are widely available but have had more limited application to food testing compared with their use in clinical laboratories. when testing food samples, false negative pcr results can occur and may be due to interference with target-cell lysis necessary for nucleic acid extraction, nucleic acid degradation and/or direct inhibition of the pcr. therefore, it is essential to include appropriate controls for the application of pcr to the detection of pathogens in f ...200717604864
a novel promoter trap identifies listeria monocytogenes promoters expressed at a low ph within the macrophage phagosome.the gram-positive pathogen listeria monocytogenes encounters acid environments in low-ph foods, during passage through the stomach and within the macrophage phagosome during systemic infection. a novel promoter-trap system termed pgad-hly was developed, based on a plasmid containing a promoterless copy of gadb (required for survival at low ph) and hly (whose product facilitates escape from the macrophage phagosome) to identify loci that are induced under different stress conditions in vitro as w ...200717608802
antimicrobial effects of a microemulsion and a nanoemulsion on enteric and other pathogens and biofilms.some microemulsions and nanoemulsions may have antimicrobial properties and be effective anti-biofilm agents. we examined the abilities of two fine emulsions, designated bctp and teop, to inactivate suspensions of vegetative cells of salmonella spp. escherichia coli 0157:h7 (vt-), pseudomonas aeruginosa, staphylococcus aureus and listeria monocytogenes. bctp is an o/w nanoemulsion of soybean oil and tri-n-butyl phosphate emulsified with triton x-100, while teop is an o/w microemulsion of ethyl o ...200717610974
effects of inoculation level, material hydration, and stainless steel surface roughness on the transfer of listeria monocytogenes from inoculated bologna to stainless steel and high-density polyethylene.the influence of inoculation level, material hydration, and stainless steel surface roughness on the transfer of listeria monocytogenes from inoculated bologna to processing surfaces (stainless steel and polyethylene) was assessed. slices of bologna (14 g) were inoculated with listeria at different levels, from 10(5) to 10(9) cfu/cm2. transfer experiments were done at a constant contact time (30 s) and pressure (45 kpa) with a universal testing machine. after transfer, cells that had been transf ...200717612072
predicting microbial heat inactivation under nonisothermal treatments.the aim of this study was to develop an equation that accurately predicts microbial heat inactivation under nonisothermal treatments at constantly rising heating rates (from 0.5 to 5 degrees c/min) in media with different ph values (4.0 or 7.4). the survival curves of all bacteria (enterococcus faecium, escherichia coli, listeria monocytogenes, salmonella senftenberg 775w, salmonella typhimurium, and staphylococcus aureus) tested under isothermal treatments were nearly linear. for the most heat- ...200717612077
low prevalence of listeria monocytogenes in foods from italy.listeria monocytogenes is an important foodborne pathogen that causes gastrointestinal disorders, and, especially in immunocompromised people, serious extraintestinal diseases, such as septicemia and meningitis, as well as abortion in pregnant women. many foods, from both plant and animal origin, have been involved in listeriosis outbreaks. this article reports the results of a 12-year survey (1993 through 2004) on the presence of l. monocytogenes in several kinds of food marketed in italy. of 5 ...200717612085
traceback identification of an ingredient (pork dewlap) as the possible source of listeria monocytogenes serotype 4b contamination in raw chicken products.in surveys conducted on finished product samples from a single poultry processing plant in spain, listeria monocytogenes was found in 14 different uncooked products. to track contamination patterns, 77 l. monocytogenes isolates were characterized by pcr-based serotyping, pulsed-field gel electrophoresis (pfge) restriction analysis, and pcr-based allelic analysis of the virulence gene acta. serotyping revealed that 12 isolates (15.6%) were of the l. monocytogenes serotype 4b complex (serotype 4b ...200717612086
combined effect of packaging atmosphere and storage temperature on growth of listeria monocytogenes on ready-to-eat shrimp.cooked, peeled, and deveined shrimp were inoculated with a 5 strain mixture of listeria monocytogenes and packaged in air, vacuum, and a 100% carbon dioxide modified atmosphere. the packaged shrimp were then stored at 3, 7, and 12 degrees c for 15 days to monitor the growth of l. monocytogenes and psychrotrophic bacteria. uninoculated shrimp were also subjected to sensory evaluation by a trained panel to measure odor and appearance over the storage period. results demonstrated that shrimp packag ...200717613367
prevalence and level of listeria monocytogenes and other listeria species in retail pre-packaged mixed vegetable salads in the uk.as part of the european commission (ec) co-ordinated programme for 2005, a study of pre-packaged ready-to-eat (rte) mixed salads containing meat or seafood ingredients from retail premises was undertaken in the uk to determine the frequency and level of listeria monocytogenes in these products. almost all (99.8%; 2682/2686) samples were of satisfactory/acceptable microbiological quality. two (0.1%) samples exceeded ec legal food safety criteria due to the presence of l. monocytogenes in excess o ...200717613368
class i/class iia bacteriocin cross-resistance phenomenon in listeria monocytogenes.variants resistant to nisin a (va), nisin z (vz), pediocin pa-1 (vp), divergicin m35 (vd) and to bacteriocin-like compounds produced by bifidobacterium thermophilum subsp. infantis rbl67 (vb) were developed from listeria monocytogenes lsd530. lactic acid production, specific growth rate, potassium ion efflux, susceptibility to 13 antibiotics, cell-envelope fatty acid composition and bacteriocin cross-resistance were evaluated. lactic acid production decreased to 75% or less of that by strain lsd ...200717613369
inactivation kinetics of inoculated escherichia coli o157:h7, listeria monocytogenes and salmonella enterica on strawberries by chlorine dioxide gas.inactivation kinetics of inoculated escherichia coli o157:h7, listeria monocytogenes and salmonella enterica on strawberries by chlorine dioxide gas at different concentrations (0.5, 1, 1.5, 3 and 5 mgl(-1)) for 10 min were studied. a cocktail of three strains of each targeted organism (100 microl) was spotted onto the surface of the strawberries (approximately 8-9 log ml(-1)) separately followed by air drying, and then treated with clo(2) gas at 22 degrees c and 90-95% relative humidity. approx ...200717613371
description of the bacteriocins produced by two strains of enterococcus faecium isolated from italian goat milk.in this study two strains of enterococcus faecium, m241 and m249, isolated from goat milk, were studied for their capability to produce antibacterial compounds. it was determined that the bacteriocins produced by both strains were active towards listeria monocytogenes and clostridium butyricum, and they did not have any activity with respect to other species of lactic acid bacteria. enterocins a and b were targeted by polymerase chain reaction (pcr) and sequenced, after cloning, in both strains. ...200717613373
growth and bacteriocin production by lactic acid bacteria in vegetable broth and their effectiveness at reducing listeria monocytogenes in vitro and in fresh-cut lettuce.the fresh-cut fruit and vegetable industry is searching for alternatives to replace chemical treatments with biopreservative approaches that ensure the safety of the product and fulfil consumer preferences for minimally processed foods. in this study, the use of bacteriocins produced by lactic acid bacteria has been tested as a substitute for chemical disinfection of fresh-cut iceberg lettuce. first, the ability of several non-plant origin bacteriocinogenic strains (nisin z(+), plantaricin c(+), ...200717613374
effect of a simple chlorine dioxide method for controlling five foodborne pathogens, yeasts and molds on blueberries.the effect of aqueous chlorine dioxide (clo(2)) on controlling foodborne pathogens, yeasts, and molds on blueberries was studied. five pathogens were spot-inoculated on the skin of blueberries. a sachet containing necessary chemicals for generation of clo(2) was used to provide 320 ppm of clo(2) in 7.5 l of water. the efficacy of different concentrations (1, 3, 5, 10, and 15 ppm) of clo(2) and various contact times (10s; 1, 5, 10, 20, 30 min; and 1h and 2h) were studied. clo(2) was most effectiv ...200717613378
active bacterial myocarditis: a case report and review of the literature.bacterial myocarditis (bm) is an uncommon cause of infectious myocarditis. bm is usually seen in the context of overwhelming sepsis or as part of a specific bacterial syndrome. the definitive diagnosis of bacterial myocarditis requires biopsy or morphologically proven active myocarditis with evidence of bacterial invasion or positive tissue cultures. the management of bacterial myocarditis consists of aggressive and early antibiotic or anti-toxin treatment, appropriate hemodynamic support, and t ...200717613408
bacteriocin-based strategies for food biopreservation.bacteriocins are ribosomally-synthesized peptides or proteins with antimicrobial activity, produced by different groups of bacteria. many lactic acid bacteria (lab) produce bacteriocins with rather broad spectra of inhibition. several lab bacteriocins offer potential applications in food preservation, and the use of bacteriocins in the food industry can help to reduce the addition of chemical preservatives as well as the intensity of heat treatments, resulting in foods which are more naturally p ...200717614151
nucleoside analogues are activated by bacterial deoxyribonucleoside kinases in a species-specific manner.to investigate the bactericidal activity of antiviral and anticancer nucleoside analogues against a variety of pathogenic bacteria and characterize the activating enzymes, deoxyribonucleoside kinases (dnks).200717615154
type i ifn induction in response to listeria monocytogenes in human macrophages: evidence for a differential activation of ifn regulatory factor 3 (irf3).listeria monocytogenes is a prototypic bacterium for studying innate and adaptive cellular immunity as well as host defense. using human monocyte-derived macrophages, we report that an infection with a wild-type strain, but not a listeriolysin o-deficient strain, of the gram-positive bacterium l. monocytogenes induces expression of ifn-beta and a bioactive type i ifn response. investigating the activation of signaling pathways in human macrophages after infection revealed that a wild-type strain ...200717617610
listeriology (1926-2007): the rise of a model pathogen.listeria monocytogenes is an ubiquitous gram positive bacterium responsible for a severe food borne disease in human and animals which has become a multifaceted model. indeed, it is a facultative intracellular bacterium that induces an amazingly rapid and sterilizing t-cell response and has been--and still is--widely used by immunologists. listeria's strategies to invade non-phagocytic cells and spread from cell to cell have been and still are outstandingly instrumental to address key questions ...200717618157
src, cortactin and arp2/3 complex are required for e-cadherin-mediated internalization of listeria into cells.listeria monocytogenes is a food-borne pathogen able to invade non-phagocytic cells. inla, a l. monocytogenes surface protein, interacts with human e-cadherin to promote bacterial entry. l. monocytogenes internalization is a dynamic process involving co-ordinated actin cytoskeleton rearrangements and host cell membrane remodelling at the site of bacterial attachment. interaction between e-cadherin and catenins is required to promote listeria entry, and for the establishment of adherens junctions ...200717627624
rapid determination of multi-locus sequence types of listeria monocytogenes by microtemperature-gradient gel electrophoresis.this report presents a new method for identifying multi-locus sequence types of listeria monocytogenes by microtemperature-gradient gel electrophoresis (mu-tgge). genomic comparison of l. monocytogenes serovar 1/2a strains egd-e and f6854 allowed selection of novel polymorphic sequences lmo0386 and lmo0428 as optimum regions for mu-tgge analysis, in addition to the previously identified lmo0297 gene. sequence analysis of a total of 48 standard strains revealed that the strains could be grouped i ...200717628727
prevalence of listeria monocytogenes in fresh and fermented italian sausages and ribotyping of contaminating strains.listeria monocytogenes has been detected in fresh as well as dry and semidry fermented sausages, rendering preparation and consumption of these products as a potential risk to human health. the aims of this study were (1) to evaluate the l. monocytogenes prevalence in 288 fresh and 237 fermented sausages produced in northern italy; (2) to quantify the average pathogen most probable number (mpn) per g of sausage; (3) to evaluate the sausage strain genetic diversity by automated pvuii ribotyping; ...200717629582
use of antimicrobial biodegradable packaging to control listeria monocytogenes during storage of cooked ham.the antimicrobial effect against l. monocytogenes of biodegradable films (alginate, zein and polyvinyl alcohol) containing enterocins was investigated. survival of the pathogen was studied by means of challenge tests performed at 6 degrees c during 8 and 29 days, for air-packed and vacuum-packed sliced cooked ham, respectively. air packaging was tested with two concentrations of enterocins (200 and 2000 au/cm2). control air-packed cooked ham showed an increase of l. monocytogenes from 10(4) to 1 ...200717629977
distribution and characteristics of listeria monocytogenes isolates from surface waters of the south nation river watershed, ontario, canada.listeria monocytogenes is a facultative intracellular pathogen thought to be widely distributed in the environment. we investigated the prevalence and characteristics of l. monocytogenes isolates from surface waters derived from catchments within the south nation river watershed (ontario, canada). this watershed is dominated by urban and rural development, livestock and crop production, and wildlife habitats. from june to november 2005, a total of 314 surface water samples were collected biweekl ...200717630309
protein synthesis in lactic acid and pathogenic bacteria during recovery from a high pressure treatment.recovery of injured bacteria after high hydrostatic pressure (hhp) treatment is a key point in food safety. in this study, protein synthesis during the recovery of meat environment bacteria listeria monocytogenes ctc1011, lactobacillus sakei 23k, l. sakei ctc494, enterococcus faecalis ctc6365 and enterococcus faecium ctc6375 after a 400 mpa hhp treatment was analyzed by two-dimensional gel electrophoresis and peptide mass fingerprinting. after 2 h recovery from hhp treatment, the four species in ...200717631981
[case of relapsing polychondritis followed by cognitive impairment].a 64-year-old man developed cognitive impairment. a neurological examination showed hypereactive tendon reflex. a cerebrospinal fluid (csf) examination showed a mild increase in both the number of mononuclear cells (14 cells/mm3) and the protein level (45 mg/dl). an electroencephalogram showed diffuse slow wave activity. brain mr images revealed multifocal lesions in the limbic area and white matter regions. he was suspected to have autoimmune encephalitis and his symptoms improved after treated ...200717633109
myd88-mediated signals induce the bactericidal lectin regiii gamma and protect mice against intestinal listeria monocytogenes infection.listeria monocytogenes is a food-borne bacterial pathogen that causes systemic infection by traversing the intestinal mucosa. although myd88-mediated signals are essential for defense against systemic l. monocytogenes infection, the role of toll-like receptor and myd88 signaling in intestinal immunity against this pathogen has not been defined. we show that clearance of l. monocytogenes from the lumen of the distal small intestine is impaired in myd88(-/-) mice. the distal ileum of wild-type (wt ...200717635956
quantitative risk assessment of listeria monocytogenes in french cold-smoked salmon: i. quantitative exposure assessment.a quantitative assessment of the exposure to listeria monocytogenes from cold-smoked salmon (css) consumption in france is developed. the general framework is a second-order (or two-dimensional) monte carlo simulation, which characterizes the uncertainty and variability of the exposure estimate. the model takes into account the competitive bacterial growth between l. monocytogenes and the background competitive flora from the end of the production line to the consumer phase. an original algorith ...200717640216
listeriolysin o expressed in a bacterial vaccine suppresses cd4+cd25high regulatory t cell function in vivo.cd4(+)cd25(high) regulatory t cells (treg) protect the host from autoimmune diseases but are also obstacles against cancer therapies. an ideal cancer vaccine would stimulate specific cytotoxic responses and reduce/suppress treg function. in this study, we showed that escherichia coli expressing listeriolysin o and ova (e. coli llo/ova) demonstrated remarkable levels of protection against ova-expressing tumor cells. by contrast, e. coli expressing ova only (e. coli ova) showed poor protection. hi ...200717641019
effect of inhibitory extracts derived from liquid smoke combined with postprocess pasteurization for control of listeria monocytogenes on ready-to-eat meats.surface pasteurization was examined in combination with low-phenolic antimicrobial extracts derived from liquid smoke to inhibit and prevent the growth of listeria monocytogenes during the shelf life of ready-to-eat meats. in preliminary trials with retail frankfurters, one smoke derivative (2-min dip) produced a 0.3-log reduction of l. monocytogenes and a 1-min in-bag pasteurization (73.9 degrees c) produced a 2.9-log reduction, whereas a combination of the two treatments produced a 5.3-log red ...200718095426
antimicrobial activity of clove and cinnamon essential oils against listeria monocytogenes in pasteurized milk.the antimicrobial activity of essential oils (eos) of cinnamon bark, cinnamon leaf, and clove against listeria monocytogenes scott a were studied in semiskimmed milk incubated at 7 degrees c for 14 days and at 35 degrees c for 24 h. the mic was 500 ppm for cinnamon bark eo and 3,000 ppm for the cinnamon leaf and clove eos. these effective concentrations increased to 1,000 ppm for cinnamon bark eo, 3,500 ppm for clove eo, and 4,000 ppm for cinnamon leaf eo when the semiskimmed milk was incubated ...200718095427
pulsed-plasma gas-discharge inactivation of microbial pathogens in chilled poultry wash water.a pulsed-plasma gas-discharge (ppgd) system was developed for the novel decontamination of chilled poultry wash water. treatment of poultry wash water in the plasma generation chamber for up to 24 s at 4 degrees c reduced escherichia coli nctc 9001, campylobacter jejuni atcc 33560, campylobacter coli atcc 33559, listeria monocytogenes nctc 9863, salmonella enterica serovar enteritidis atcc 4931, and s. enterica serovar typhimurium atcc 14028 populations to non-detectable levels (< or = 8 log cfu ...200718095434
comparing uncertainty resulting from two-step and global regression procedures applied to microbial growth models.two different microbial modeling procedures were compared and validated against independent data for listeria monocytogenes growth. the most generally used method is two consecutive regressions: growth parameters are estimated from a primary regression of microbial counts, and a secondary regression relates the growth parameters to experimental conditions. a global regression is an alternative method in which the primary and secondary models are combined, giving a direct relationship between exp ...200718095435
different enrichment procedures for recovery of listeria monocytogenes from raw chicken samples can affect the results of detection (by chromogenic plating or real-time pcr) and lineage or strain identification.this study aimed to evaluate the effect of different enrichment procedures on the detection of listeria monocytogenes in food, by a comparison of subculture onto chromogenic agar with real-time pcr. two different culture media, the primary and secondary enrichment broths of the u.s. food safety and inspection service (fsis) method used for pcr detection of l. monocytogenes, were compared for the primary enrichment of retail ground chicken samples. l. monocytogenes was detected after the completi ...200718095442
bacterial contamination of sago starch in papua new guinea.sago starch is an important food in lowland papua new guinea. extraction of the starch from the palm and storage were performed by way of traditional methods that have been used for thousands of years. currently, very little is known about the microbiology of sago starch. sago samples were collected from areas of high starch utilization and analyzed for the presence of bacterial pathogens and indicator organisms. storage methods and duration were recorded at the time of collection, and ph and wa ...200718095446
cleaning and handling implements as potential reservoirs for bacterial contamination of some ready-to-eat foods in retail delicatessen environments.this study assessed the association of bacteria with cleaning tools, such as floor mops (n = 25) and cleaning cloths (n = 39), and handling devices, such as disposable plastic gloves (n = 20), used during filled baguette and assorted salad preparation in four selected retail delicatessens in johannesburg, south africa. samples of each cleaning or handling tool were prepared for aerobic (apc), coliform (cc), escherichia coli (ec), bacillus cereus (bcc), and staphylococcus aureus (sac) counts, as ...200718095448
post-colonoscopic listeria septicemia in ulcerative colitis during immunosuppressive therapy.a 78-year-old man who had been diagnosed with ulcerative colitis was admitted because of uncontrolled severe, frequent, bloody diarrhea. he was treated with immunosuppressive therapy that included corticosteroid and azathioprine. colonoscopy was used to assess disease activity. this revealed that the mucosa of his digestive tract from the rectum to the ileum was damaged. he developed a high-grade fever soon after colonoscopy. blood culture demonstrated listeria monocytogenes. treatment was chang ...200718084128
mapping quantitative trait loci from a single-tail sample of the phenotype distribution including survival data.a new effective bayesian quantitative trait locus (qtl) mapping approach for the analysis of single-tail selected samples of the phenotype distribution is presented. the approach extends the affected-only tests to single-tail sampling with quantitative traits such as the log-normal survival time or censored/selected traits. a great benefit of the approach is that it enables the utilization of multiple-qtl models, is easy to incorporate into different data designs (experimental and outbred popula ...200718073434
physical properties of nisin-incorporated gelatin and corn zein films and antimicrobial activity against listeria monocytogenes.edible films of gelatin and corn zein were prepared by incorporating nisin to the film-forming solutions. corn zein film with nisin of 12,000 iu/ml had an increase of 11.6 mpa in tensile strength compared with the control, whereas gelatin film had a slight increase with the increase of nisin concentration added. water vapor permeability for both corn zein and gelatin films decreased with the increase of nisin concentration, thus providing a better barrier against water. antimicrobial activity ag ...200718050958
morphological changes induced in listeria monocytogenes v7 by a bacteriocin produced by pediococcus acidilactici.pediococcus acidilactici produces bacteriocin, which kills listeria monocytogenes. the bactericidal mode of action of the bacteriocin against l. monocytogenes v7 was investigated by transmission electron microscopy. the bacteriocin was purified partially from the cell-free extract using micro-cel and cation-exchange chromatography, and the specific activity was increased 1,791 fold. the bacteriocin (6,400 au/ ml) was inoculated with l. monocytogenes v7 and incubated for 0.5 h, 1 h, 3 h, and 6 h. ...200718051280
production of monoclonal antibody against listeria monocytogenes and its application to immunochromatography strip test.an immunochromatography (icg) strip test based on a monoclonal antibody for the rapid detection of l. monocytogenes in meat and processed-meat samples was developed in this study. a monoclonal antibody (mab) specific to l. monocytogenes was produced from cloned hybridoma cells (fklm-3b12-37) and used to develop an icg strip test. the antibody showed a stronger binding to l. monocytogenes than other listeria species, and a weak cross-reaction to s. aureus based on an elisa. the detection limit of ...200718051327
high-throughput sequence alignment using graphics processing units.the recent availability of new, less expensive high-throughput dna sequencing technologies has yielded a dramatic increase in the volume of sequence data that must be analyzed. these data are being generated for several purposes, including genotyping, genome resequencing, metagenomics, and de novo genome assembly projects. sequence alignment programs such as mummer have proven essential for analysis of these data, but researchers will need ever faster, high-throughput alignment tools running on ...200718070356
defective t cell development and function in the absence of abelson kinases.thymocyte proliferation, survival, and differentiation are tightly controlled by signaling from the pre-tcr. in this study, we show for the first time that the abelson (abl) kinases regulate proximal signaling downstream of the pre-tcr. conditional deletion of abl kinases in thymocytes reveals a cell-autonomous role for these proteins in t cell development. the conditional knockout mice have reduced numbers of thymocytes, exhibit an increase in the percentage of the cd4(-)cd8(-) double-negative ...200718025176
activation of immature hepatic nk cells as immunotherapy for liver metastatic disease.nk cells can identify and eliminate emerging tumors due to altered expression of activating and inhibitory ligands on aberrant cells, a process that is greatly enhanced following nk cell activation. as a principal site of both tumor metastases and immature nk cells, the liver represents a unique anatomic location in which activation of the innate immune system could provide substantial therapeutic benefit. we describe here the nk cell-dependent destruction of a primary hepatic tumor following in ...200718025181
extensive characterization of ifn-induced gtpases mgbp1 to mgbp10 involved in host defense.ifn-gamma orchestrates a potent antimicrobial host response. however, the underlying molecular basis for this immunological defense system is largely unknown. in a systematic approach to identify ifn-gamma-regulated host effector molecules, a notable number of transcripts with consensus gtp-binding motives were obtained. further extensive transcriptome and genome analyses identified five novel family members of murine guanylate-binding proteins (mgbps) now designated mgbp6, 7, 8, 9, and 10. more ...200718025219
production of nisin z by lactococcus lactis isolated from dahi.lactococcus lactis cm1, an isolate from homemade "dahi," a traditional fermented milk from india, used maltose as carbon source to produce a high level of bacteriocin. the bacterial cell mass and the bacteriocin production correlated with the initial ph of the medium and were highest when the initial ph was 11.0. the level of bacteriocin reached its peak at the late log phase with concomitant reduction of culture ph to 4.2, regardless of the initial ph of the medium. a combination of maltose and ...200718025595
characteristics and frequency of detection of fecal listeria monocytogenes shed by livestock, wildlife, and humans.listeria monocytogenes is a facultative intracellular pathogen that can be carried asymptomatically in various animals and can be shed in feces. we investigated the prevalence and characteristics of l. monocytogenes isolated from livestock, wildlife, and human potential sources of contamination in 2 areas in ontario, canada. from february 2003 to november 2005, a total of 268 fecal samples were collected from different animals. listeria monocytogenes was isolated using selective enrichment, isol ...200718026208
neuroinfections due to listeria monocytogenes.listeria monocytogenes is not a rare pathogen causing meningitis, mainly in small children and in close contacts to livestock. the pathogen is naturally resistant to cephalosporins and some glycopeptides as well, therefore despite of syndromologic diagnosis of meningitis and initial therapy with 3rd generation cephalosporins according to the guidelines therapeutic failures with clinical consequences may occur.200718030269
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