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evidence for the role of horizontal transfer in generating pvt1, a large mosaic conjugative plasmid from the clam pathogen, vibrio tapetis.the marine bacterium vibrio tapetis is the causative agent of the brown ring disease, which affects the clam ruditapes philippinarum and causes heavy economic losses in north of europe and in eastern asia. further characterization of v. tapetis isolates showed that all the investigated strains harbored at least one large plasmid. we determined the sequence of the 82,266 bp plasmid pvt1 from the cect4600(t) reference strain and analyzed its genetic content. pvt1 is a mosaic plasmid closely relate ...201121326607
enumeration of vibrio parahaemolyticus in the viable but nonculturable state using direct plate counts and recognition of individual gene fluorescence in situ hybridization.vibrio parahaemolyticus is a gram-negative, halophilic bacterium indigenous to marine and estuarine environments and it is capable of causing food and water-borne illness in humans. it can also cause disease in marine animals, including cultured species. currently, culture-based techniques are used for quantification of v. parahaemolyticus in environmental samples; however, these can be misleading as they fail to detect v. parahaemolyticus in a viable but nonculturable (vbnc) state which leads t ...201121329738
detecting potentially virulent vibrio vulnificus strains in raw oysters by quantitative loop-mediated isothermal amplification.vibrio vulnificus is a leading cause of seafood-related deaths in the united states. sequence variations in the virulence-correlated gene (vcg) have been used to distinguish between clinical and environmental v. vulnificus strains, with a strong association between clinical ones and the c sequence variant (vcgc). in this study, vcgc was selected as the target to design a loop-mediated isothermal amplification (lamp) assay for the rapid, sensitive, specific, and quantitative detection of potentia ...201121357428
tumor necrosis factor-a and mortality in patients infected with vibrio vulnificus.serum tumor necrosis factor-a (tnf-a) was evaluated in vibrio vulnificus-infected patients at admission. the median tnf-a concentration in the non-survivor group was determined to be 261.0 pg/ml, in contrast to 69.5 pg/ml in the survivor group (p = 0.001). hence, serum tnf-a concentration may potentially be an early predictor of the mortality in patients with vibrio septicemia.201121363981
vibrio vulnificus oysters: pearls and perils.vibrio vulnificus is a naturally occurring bacterium found ubiquitously in coastal waters. infection with this organism, which is often associated with eating raw oysters, is the leading cause of seafood-related deaths in the united states. this article reviews the current scientific literature on this potentially lethal pathogen and discusses the prognosis, treatment, and prevention of v. vulnificus infections.201121367733
involvement of the gspab complex in assembly of the type ii secretion system secretin of aeromonas and vibrio species.the type ii secretion system (t2ss) functions as a transport mechanism to translocate proteins from the periplasm to the extracellular environment. the exea homologue in aeromonas hydrophila, gspa(ah), is an atpase that interacts with peptidoglycan and forms an inner membrane complex with the exeb homologue (gspb(ah)). the complex may be required to generate space in the peptidoglycan mesh that is necessary for the transport and assembly of the megadalton-sized exed homologue (gspd(ah)) secretin ...201121378198
bullae and sweat gland necrosis in the differential diagnosis for vibrio vulnificus infection in an alcoholic patient.bullae and sweat gland necrosis remain rare cutaneous manifestation, and these conditions can be misdiagnosed as vibrio vulnificus infections or other soft tissue infections because of their low index of suspicion. a 46-yr-old man with a history of continued alcohol consumption presented with erythematous and hemorrhagic bullous lesions on his left arm. the patient reported that after the ingestion of clams, he slept for 12 hr in a heavily intoxicated state. then the skin lesions started as a re ...201121394318
antimicrobial effects of essential oils of cinnamosma fragrans on the bacterial communities in the rearing water of penaeus monodon larvae.farmed shrimps are vectors of various vibrio species that are considered a potential health hazard. previous study has shown that vibrio parahaemolyticus, vibrio vulnificus, and vibrio alginolyticus dominated in the water and larval samples of shrimp hatchery (randrianarivelo et al. 2010 ). the effects of two essential oils (eos) of cinnamosma fragrans, an endemic plant to madagascar (b8: linalool-type and b143: 1,8-cineole-type), were determined on the total heterotrophic aerobic bacteria and t ...201121395412
structure of a nadph-dependent blue fluorescent protein revealed the unique role of gly176 on the fluorescence enhancement.a nadph-dependent blue fluorescent protein from vibrio vulnificus ckm-1 (bfpvv) emits blue fluorescence under uv-exposure. previously, the bfpvvd7 mutant generated by directed evolution displayed a fourfold enhancement in fluorescent intensity. herein, a further increase in fluorescence in the new bfpvvd8 mutant, with three additional mutations from bfpvvd7, was made. to understand the underlying mechanism of the increased fluorescent intensity of bfpvv, we solved the bfpvvd8-nadph complex struc ...201121397029
regulation of cytotoxicity by quorum-sensing signaling in vibrio vulnificus is mediated by smcr, a repressor of hlyu.cytotoxicity is an important virulence determinant in the pathogenesis of vibrio vulnificus, and two cytotoxins, rtx (encoded by rtxa1) and cytolysin/hemolysin (encoded by vvha), have been identified in this organism. we showed that the quorum-sensing regulator luxo controlled the cytotoxicity of this organism: a +öluxo mutant exhibited low cytotoxicity, whereas a constitutively activated luxo mutant, luxo(d47e), remained highly cytotoxic. the cytotoxicity of the +öluxo mutant was restored when ...201121398530
high intragenomic heterogeneity of 16s rrna genes in a subset of vibrio vulnificus strains from the western mediterranean coast.heterogeneity among ribosomal operons in vibrio vulnificus is purported as a probabilistic indicator of strain virulence and classifies v. vulnificus strains as 16s rrna genes type a and b. in this study, 16s rrna genes typing of v. vulnificus strains isolated from the valencia city coast, in the western mediterranean, showed that 24 out of 30 isolates were type a, one was type b and five could not be typed. single strand conformation polymorphism (sscp) analysis of this gene region revealed com ...201021404212
rtx toxin enhances the survival of vibrio vulnificus during infection by protecting the organism from phagocytosis.vibrio vulnificus is a marine bacterium causing serious septicemia and wound infection in humans. it produces an rtx toxin that can lyse a variety of cells and is important for virulence in mice. in this study, we explored the role of rtx in pathogenesis by characterizing an rtx-deficient mutant. this mutant showed an ~2-log reduction in virulence for mice infected by various routes. survival of the mutant at the infection site and subsequent spread into the bloodstream were impaired. in mice pr ...201121422475
inactivation of vibrio vulnificus hemolysin through mutation of the n- or c-terminus of the lectin-like domain.vibrio vulnificus is an etiological agent causing serious systemic infections in the immunocompromised humans or cultured eels. this species commonly produces a hemolytic toxin consisting of the cytolysin domain and the lectin-like domain. for hemolysis, the lectin-like domain specifically binds to cholesterol in the erythrocyte membrane, and to form a hollow oligomer, the toxin is subsequently assembled on the membrane. the cytolysin domain is essential for the process to form the oligomer. thr ...201121426913
in-vitro antagonistic characteristics of crude aqueous and methanolic extracts of garcinia kola (heckel) seeds against some vibrio bacteria.the methanolic and aqueous extracts of garcinia kola seeds were screened for their anti-vibrio activities against 50 vibrio isolates obtained from wastewater final effluents in the eastern cape province, south africa. the crude extracts at 10 mg/ml exhibited appreciable inhibitory activities against most of the test vibrio isolates, with zones of inhibition ranging from 10-19 mm for methanol extract and 8-15 mm for the aqueous extracts. the minimum inhibitory concentrations (mic) of the methanol ...201121441874
dynamic analysis of pathogen-infected host cells using quantitative phase microscopy.we present the real-time quantitative analysis of vibrio vulnificus-infected host cells using quantitative phase microscopy (qpm) based on interferometric techniques. this provides the ability to retrieve the phase or optical path-length distribution over the cell with nanometer path-length sensitivity from a single interferogram image. we have used qpm to study dynamic cell morphologic changes and to noninvasively quantify the cell volumes of rat basophilic leukemia rbl-2h3 cells infected with ...201121456868
proteases produced by vibrios.bacteria of the genus vibrio are normal habitants of the aquatic environment but the some species are believed to be human pathogens. pathogenic vibrios produce various pathogenic factors, and the proteases are also recognized to play pathogenic roles in the infection: the direct roles by digesting many kinds of host proteins or indirect roles by processing other pathogenic protein factors. especially vvp from vibrio vulnificus is thought to be a major pathogenic factor of the vibrio. although h ...201121467624
factors affecting mortality in hong kong patients with upper limb necrotising fasciitis.to identify predictive factors for mortality of patients with upper limb necrotising fasciitis.201121471588
bacterial pathogens in hawaiian coastal streams-associations with fecal indicators, land cover, and water quality.this work aimed to understand the distribution of five bacterial pathogens in o'ahu coastal streams and relate their presence to microbial indicator concentrations, land cover of the surrounding watersheds, and physical-chemical measures of stream water quality. twenty-two streams were sampled four times (in december and march, before sunrise and at high noon) to capture seasonal and time of day variation. salmonella, campylobacter, staphylococcus aureus, vibrio vulnificus, and v. parahaemolytic ...201121492899
a survey of oysters (crassostrea gigas) in new zealand for vibrio parahaemolyticus and vibrio vulnificus.a microbiological survey was conducted to determine the levels of total and pathogenic vibrio parahaemolyticus (vp) and vibrio vulnificus (vv) in pacific oysters (crassostrea gigas) collected from commercial growing areas in the north island, new zealand. the survey was intended to be geographically representative of commercial growing areas of pacific oysters in new zealand, while selecting the time frame most likely to coincide with the increased abundance of pathogenic vibrio species. vp was ...201121501884
rapid and sensitive detection of vibrio vulnificus by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to rpos gene.a novel loop-mediated isothermal amplification (lamp) combined with amplicon detection by chromatographic lateral flow dipstick (lfd) assay was developed and evaluated for the detection of vibrio vulnificus. biotinylated lamp amplicons were produced by a set of six designed primers that recognized the v. vulnificus rna polymerase subunit sigma factor s (rpos) gene followed by hybridization with an fitc-labeled probe and lfd detection. the optimized time and temperature conditions for the lamp as ...201121513793
comparison of the effects of environmental parameters on growth rates of vibrio vulnificus biotypes i, ii and iii by culture and quantitative pcr.vibrio vulnificus is a natural inhabitant of estuarine waters. the three known biotypes include 1 (most human pathogens), 2 (primarily eel pathogens), and 3 (associated with fish and with human wound infections in israel). despite the frequently lethal consequences of v. vulnificus infections, the growth rates of the various biotypes and their response to environmental changes are not well characterized. we compared the specific growth rates (μ) of a representative of each biotype by culture and ...201121515718
multiplex pcr assay for identification of three major pathogenic vibrio spp., vibrio cholerae, vibrio parahaemolyticus, and vibrio vulnificus.a multiplex pcr assay was developed based on atpa-sequence diversification for molecular identification of 3 major pathogenic vibrio species: vibrio cholerae, vibrio parahaemolyticus, and vibrio vulnificus. it specifically identified them from among 133 strains of various vibrio species and other genera, and was applicable for testing seawater, suggesting its usefulness.201121530641
modulation of iron-uptake systems by a mutation of luxs encoding an autoinducer-2 synthase in vibrio vulnificus.vibrio vulnificus possesses multiple iron-uptake systems which are mediated by vuua (vulnibactin receptor), iuta (aerobactin receptor) and hupa (heme receptor). in this study, we determined the effect of a mutation of luxs encoding autoinducer-2 (ai-2) synthase on the expressions of the three receptors. a mutation and an in trans complementation of luxs did not affect the growing ability of v. vulnificus in iron-deficient conditions. nevertheless, the luxs mutation slightly decreased vuua expres ...201121532149
inactivation kinetics of vibrio vulnificus in phosphate-buffered saline at different freezing and storage temperatures and times.vibrio vulnificus (vv) is a pathogen that can be found in raw oysters. freezing can reduce vv and increase the shelf life of oysters. the objective of this study was to develop predictive inactivation kinetic models for pure cultures of vv at different frozen storage temperatures and times. vv was diluted in phosphate-buffered saline (pbs) to obtain about 10(7) cfu/ml. samples were frozen at -10, -35, and -80 °c (different freezing rates), and stored at different temperatures. survival of vv was ...201121535763
the pseudomonas quinolone signal (pqs), and its precursor hhq, modulate interspecies and interkingdom behaviour.the pseudomonas quinolone signal (pqs), and its precursor 2-heptyl-4-quinolone (hhq), play a key role in coordinating virulence in the important cystic fibrosis pathogen pseudomonas aeruginosa. the discovery of hhq analogues in burkholderia and other microorganisms led us to investigate the possiblity that these compounds can influence interspecies behaviour. we found that surface-associated phenotypes were repressed in gram-positive and gram-negative bacteria as well as in pathogenic yeast in r ...201121539583
vibrio parahemolyticus septicaemia in a liver transplant patient: a case report.abstract: introduction: vibrio parahemolyticus is the leading cause of vibrio-associated gastroenteritis in the united states of america, usually related to poor food handling; only rarely has it been reported to cause serious infections including sepsis and soft tissue infections. in contrast, vibrio vulnificus is a well-known cause of septicaemia, especially in patients with cirrhosis. we present a patient with v. parahemolyticus sepsis who had an orthotic liver transplant in 2007 and was on i ...201121548914
a conserved gtpase ychf of vibrio vulnificus is involved in macrophage cytotoxicity, iron acquisition, and mouse virulence.vibrio vulnificus, a highly virulent marine bacterium, causes serious wound infections and fatal septicemia in many areas of the world. to identify v. vulnificus genes required for killing macrophages, we made an insertional mutant library of v. vulnificus and screened it for reduced macrophage cytotoxicity. one mutant defective in macrophage cytotoxicity had an insertion in ychf, a gene encoding a putative gtpase. in addition to reduced cytotoxicity, this mutant had attenuated growth in iron-li ...201121570909
[development of an immunochromatographic strip test for rapid detection of vibrio vulnificus.]objective: to establish a rapid strip test for detection of vibrio vulnificus. method: anti-vibrio vulnificus polyclonal antibodies were obtained from the rabbits immunized with vibrio vulnificus (atcc27562). colloidal gold was prepared through reducing haucl(4)·4h(2)o by sodium citrate and conjugated with the polyclonal antibodies as the detecting reagent. the polyclonal antibodies and sheep anti-rabbit immunoglobulins were separately coated onto the same nitrocellulose membrane for sample dete ...201121602152
frsa functions as a cofactor-independent decarboxylase to control metabolic flux.the interaction between fermentation-respiration switch (frsa) protein and glucose-specific enzyme iia(glc) increases glucose fermentation under oxygen-limited conditions. we show that frsa converts pyruvate to acetaldehyde and carbon dioxide in a cofactor-independent manner and that its pyruvate decarboxylation activity is enhanced by the dephosphorylated form of iia(glc) (d-iia(glc)). crystal structures of frsa and its complex with d-iia(glc) revealed residues required for catalysis as well as ...201121623357
transgenic expression of tilapia hepcidin 1-5 and shrimp chelonianin in zebrafish and their resistance to bacterial pathogens.recently, tilapia hepcidin (th)1-5 was characterized, and its antimicrobial functions against several pathogens were reported. the antimicrobial functions of another shrimp antimicrobial peptide (amp), chelonianin, were also characterized using a recombinant chelonianin protein (rcf) that was expressed by a stably transfected chinese hamster ovary (cho) cell line against pathogen infections in fish. the function of the overexpression of both amps in zebrafish muscles was not examined in previous ...201121642002
temporal and spatial variability in culturable pathogenic vibrio spp. in lake pontchartrain, louisiana, usa, following hurricanes katrina and rita.we investigated the abundance, distribution, and virulence gene content of v. cholerae, v. parahaemolyticus, v. vulnificus in the waters of southern lake pontchartrain in louisiana, usa, on four occasions from october 2005 to september 2006, using selective cultivation and molecular assays. the three, targeted pathogenic vibrios were generally below detection in january 2006 when the water was cold (13° c), and highest in september 2006 when lake water was warmest (30° c). the maximum values for ...201121642406
identification and characterization of a novel serine protease vvps containing two functional domains and essential for autolysis of vibrio vulnificus.little is known about the molecular mechanism for autolysis of gram (-) bacteria. in the present study, we identified the vvps gene encoding a serine protease, vvps, from vibrio vulnificus, a gram (-) food-born pathogen. the amino acid sequence predicted that vvps consists of two functional domains, an n-terminal protease catalytic domain (pcd) and c-terminal carbohydrate binding domain (cbd). a null mutation of vvps significantly enhanced viability during stationary phase, as measured by enumer ...201121642466
design and validation of a novel multiplex real-time pcr assay for vibrio pathogen detection.three species--vibrio cholerae, vibrio parahaemolyticus, and vibrio vulnificus--account for the majority of vibrio infections in humans. rapid and accurate identification of vibrio species has been problematic because phenotypic characteristics are variable within species. additionally, biochemical identification and confirmation require 2 or more days to complete. rapid and sensitive molecular techniques for the detection of vibrio pathogens would be useful for the surveillance and management o ...201121669071
chitin-induced carbotype conversion in vibrio vulnificus.as an etiological agent of bacterial sepsis and wound infections, vibrio vulnificus is unique among the vibrionaceae. the most intensely studied of its virulence factors is the capsular polysaccharide (cps). over 100 cps types have been identified, yet little is known about the genetic mechanisms that drive such diversity. chitin, the second-most-abundant polysaccharide in nature, is known to induce competence in vibrio species. here, we show that the frequency of chitin-induced transformation i ...201121670169
necrotising fasciitis caused by vibrio vulnificus in the lower limb following exposure to seafood on the hand.vibrio vulnificus infection mainly manifests as primary bacteraemia or gastroenteritis following injection of the microorganism, and wound infection through direct inoculation. injury from exposure to the microorganism and development of necrotising fasciitis at a remote site and sepsis are rare. this report is of a high-risk patient with haemoglobin h disease who developed necrotising fasciitis in the lower limb after exposure to seafood on the hand.201121813906
functional characterization of epsc, a component of the type ii secretion system, in the pathogenicity of vibrio vulnificus.epsc, one of the components comprising the type ii secretion system (t2ss), was isolated from a human pathogenic bacterium, vibrio vulnificus, to evaluate its role in eliciting virulence. an espc-deleted mutant v. vulnificus displayed a reduced cytotoxicity to the human cell line hep-2 and an attenuated virulence in a mouse model. this mutant exhibited dramatic defects in secretion of diverse extracellular proteins, such as outermembrane proteins, transporters, and the known secreted factors, no ...201121788383
a new leukocyte cell-derived chemotaxin-2 from marine fish grouper, epinephelus coioides: molecular cloning and expression analysis.leukocyte cell-derived chemotaxin-2 (lect2) is a multifunctional protein involved in cell growth, differentiation and autoimmunity. in this study, a new leukocyte cell-derived chemotaxin-2 (eclect2) gene was cloned from grouper, epinephelus coioides, by rapid amplification of cdna ends (race) pcr. the full-length cdna sequence of eclect2 was 595 bp in size, containing a 5'-untranslated region (utr) of 44 bp and a 3'-utr of 83 bp. the deduced protein sequence of the open reading frame (465 bp) sh ...201121763428
insights into the antibacterial and immunomodulatory functions of tilapia hepcidin (th)2-3 against vibrio vulnificus infection in mice.the antimicrobial and immunomodulatory functions of the antimicrobial peptide, tilapia hepcidin (th)2-3, against a bacterial endotoxin under in vitro conditions was previously reported. in this study, we investigated the antibacterial and immunomodulatory functions of th2-3 in mice infected with the pathogen, vibrio vulnificus. a th2-3 injection in v. vulnificus-infected mice produced an increased survival rate compared to mice injected with v. vulnificus only. in addition, a th2-3 injection inc ...201121756935
the antifouling potentiality of galactosamine characterized from vibrio vulnificus exopolysaccharide.to gain a better insight into biofilm composition, the exopolysaccharide (eps) of the gram-negative bacterium vibrio vulnificus was studied. monosaccharide composition analysis of the wild-type and mutant v. vulnificus eps carried out with bio-liquid chromatography revealed the presence of d-glucosamine, d-galactose, d-glucose and d-xylose in both strains. d-galactosamine was found only in the mutant that formed less biofilm compared to its wild-type. the influence of galactosamine on biofilm fo ...201121827336
comparative analysis of proteins in the culture supernatants of human intestinal epithelial cells infected with the wild-type and rtxe mutant of vibrio vulnificus.bacterial virulence factors and secreted extracellular proteins from damaged host cells following infection have been recognized as key mediators in the pathophysiological alterations observed in septic shock, and have also been shown to have a synergistic influence on bacterial pathogenicity. we hypothesized that during infections, virulence factors as well as host-shed proteins may synergistically influence aspects of the pathogenicity of v.-ávulnificus, such as primary septic shock and overpr ...201121725587
influence of iron-chelated growth conditions on outer membrane protein production and virulence of vibrio tubiashii.growth of two vibrio tubiashii strains under iron-chelated conditions resulted in the production of a hydroxymate-like siderophore, and expression of outer membrane proteins with homologies to proteins in vibrio cholerae and vibrio vulnificus which were not seen in cells grown under non-chelated growth conditions. pcr analysis using primers based on listonella anguillarum's ferric uptake repressor protein (fur) gene detected a 316-ábp fur gene homolog which also had sequence homology to the fur ...201121839393
genomic and metabolic profiling of nonulosonic acids in vibrionaceae reveal biochemical phenotypes of allelic divergence in vibrio vulnificus.nonulosonic acids (nulos) encompass a large group of structurally diverse nine-carbon backbone +¦-keto sugars widely distributed among the three domains of life. mammals express a specialized version of nulos called sialic acids, which are displayed in prominent terminal positions of cell surface and secreted glycoconjugates. within bacteria, the ability to synthesize nulos has been demonstrated in a number of human pathogens and is phylogenetically widespread. here we examine the distribution, ...201121724895
intranasal immunization with recombinant pspa fused with a flagellin enhances cross-protective immunity against streptococcus pneumoniae infection in mice.streptococcus pneumoniae is a major respiratory pathogen that causes high levels of mortality and morbidity in infants and the elderly. despite the use of antibiotics and vaccines, fatal pneumococcal disease remains prevalent. pneumococcal surface protein a (pspa), a highly immunogenic surface protein produced by all strains of s. pneumoniae, can elicit protective immunity against fatal pneumococcal infection. we have previously demonstrated that the vibrio vulnificus flab, a bacterial flagellin ...201121696869
occurrence and potential pathogenesis of vibrio cholerae, vibrio parahaemolyticus and vibrio vulnificus on the south coast of sweden.during the summer of 2006, several wound infections - of which three were fatal - caused by vibrio cholerae were reported from patients who had been exposed to water from the baltic sea. before these reports, we initiated a sampling project investigating the occurrence of potential human pathogenic v. cholerae, vibrio vulnificus and vibrio parahaemolyticus in the sound between sweden and denmark. the blue mussel (mytilus edulis) was used as an indicator to follow the occurrence of vibrios over t ...201121692819
Vibrio virulence genes in fishes collected from estuarine waters in Italy.Aims:ÔÇé The aim of this study was to investigate the presence of Vibrio vulnificus and potentially pathogenic strains of Vibrio parahaemolyticus in mullets collected from estuarine environment in Italy. Methods and Results:ÔÇé Two hundred and ninety-five mullets were analysed by culture using the selective medium thiosulfate citrate bile salt sucrose agar, during a monitoring period of 2ÔÇâyears (2008-2009). Presumptive Vibrio colonies were initially identified by using biochemical tests, and s ...201121740448
Detection and characterization of pathogenic vibrios in shellfish by a Ligation Detection Reaction-Universal Array approach.Vibrios are a group of major foodborne pathogens widely distributed in marine environment. Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus are the pathogenic species of Vibrio that pose the greatest threat to human health. However, other vibrios, e.g. Vibrio alginolyticus, Vibrio mimicus and Grimontia hollisae, apparently less relevant in the group of foodborne pathogens, have been sporadically found in outbreaks. For seafood safety and economic purposes, a rapid and powerful met ...201122177227
Intranasal administration of a flagellin-adjuvanted inactivated influenza vaccine enhances mucosal immune responses to protect mice against lethal infection.The influenza virus, a mucosal pathogen that infects the respiratory tract, is a major global health issue. There have been attempts to mucosally administer inactivated influenza vaccines to induce both mucosal and systemic immune responses. However, mucosally administered inactivated influenza vaccine has low immunogenicity, which is partially due to the lack of an effective mucosal adjuvant. The development of a safe and effective mucosal adjuvant is a prerequisite to the practical use of a mu ...201222051136
Insights into the antibacterial and immunomodulatory functions of the antimicrobial peptide, epinecidin-1, against Vibrio vulnificus infection in zebrafish.In the present study, we used Vibrio vulnificus and a zebrafish model system to investigate the inhibitory effect of epinecidin-1 on acute bacterial infection and studied the impacts of pretreatment, co-treatment, and post-treatment with epinecidin-1 on its protective efficacy. In vivo experiments showed that co-treatment with epinecidin-1 and V. vulnificus achieved 78%-97% survival rates after 30 days. When epinecidin-1 and V. vulnificus were co-injected into zebrafish and zebrafish were re-cha ...201121925271
Seagrass as a potential source of natural antioxidant and anti-inflammatory agents.Context: Halophila spp. is a strong medicine against malaria and skin diseases and is found to be very effective in early stages of leprosy. Seagrasses are nutraceutical in nature and therefore of importance as food supplements. Objective: The antibacterial, antioxidant, and anti-inflammatory activities of Halophila ovalis R. Br. Hooke (Hydrocharitaceae) methanol extract were investigated and the chemical constituents of purified fractions were analyzed. Materials and methods: Plant material ...201122129224
characterization of a new quorum sensing regulator luxt and its roles in the extracellular protease production, motility, and virulence in fish pathogen vibrio alginolyticus.vibrio alginolyticus, an opportunistic pathogen that causes vibriosis in miscellaneous fish species, has brought about serious economic damage to the mariculture industry in south china. the mechanism of virulence regulation in v. alginolyticus is yet not known except a vibrio harveyi-like quorum sensing (qs) system that is established to manipulate the expression of diverse genes including those encoding virulence determinants. in this study, a new tetr family qs regulator, luxt, was identified ...201122130678
the effect of metals on condition and pathologies of european eel (anguilla anguilla): in situ and laboratory experiments.forty-nine wild eels (anguilla anguilla) caught in the albufera lake (spain), measuring 24.0-75.0cm in length and 25.0-637.7g in weight, were examined for metals (cd, co, cr, cu, fe, hg, mn, pb, se and zn), condition (ci and hsi indices), as well as for diseases (anguillicola infestation; bacterial infections). total metal load significantly increased in eel liver tissue parallel to total length and body weight (log), while silvering females (w(b)>200g; l≥500mm) exhibited the highest amounts of ...201122030412
purification and characterization of enterocin mc13 produced by a potential aquaculture probiont enterococcus faecium mc13 isolated from the gut of mugil cephalus.a bacteriocin producer strain mc13 was isolated from the gut of mugil cephalus (grey mullet) and identified as enterococcus faecium. the bacteriocin of e. faecium mc13 was purified to homogeneity, as confirmed by tricine sodium dodecyl sulphate - polyacrylamide gel electrophoresis (sds-page). reverse-phase high-performance liquid chromatography (hplc) analysis showed a single active fraction eluted at 26 min, and matrix-assisted laser desorption ionization time of flight (maldi-tof) mass spectro ...201122112158
Overlapping and unique contributions of two conserved polysaccharide loci in governing distinct survival phenotypes in Vibrio vulnificus.As an aetiological agent of bacterial sepsis and wound infections, Vibrio vulnificus is unique among the Vibrionacea. Its continued environmental persistence and transmission are bolstered by its ability to colonize shellfish and form biofilms on various marine biotic surfaces. We previously identified a polysaccharide locus, brp, which contributes to the survival phenotypes of biofilm formation, rugose colony formation and stress resistance. Here, we describe a second polysaccharide locus, rbd ...201121895917
Prevalence and antimicrobial susceptibility of major foodborne pathogens in imported seafood.Seafood is a leading commodity implicated in foodborne disease outbreaks in the United States. Seafood importation rose dramatically in the past 3 decades and now contributes to more than 80% of the total U.S. seafood supply. However, limited data are available on the microbiological safety of imported seafood. In this study, we obtained a total of 171 salmon, shrimp, and tilapia samples imported from 12 countries in three retail stores in Baton Rouge, LA. The total microbial population and the ...201121902913
High salinity relay as a postharvest processing strategy to reduce vibrio vulnificus levels in Chesapeake Bay oysters (Crassostrea virginica).In 2009 the U.S. Food and Drug Administration (FDA) announced its intention to implement postharvest processing (PHP) methods to eliminate Vibrio vulnificus from oysters intended for the raw, half-shell market that are harvested from the Gulf of Mexico during warmer months. FDA-approved PHP methods can be expensive and may be associated with unfavorable responses from some consumers. A relatively unexplored PHP method that uses relaying to high salinity waters could be an alternative strategy, ...201122054191
Identification and characterization of a novel outer membrane protein receptor required for hemin utilization in Vibrio vulnificus.Vibrio vulnificus, the cause of septicemia and serious wound infection in humans and fishes, require iron for its pathogenesis. Hemin uptake through the outer membrane receptor, HupA, is one of its many mechanisms by which it acquires iron. We report here the identification of an additional TonB-dependent hemin receptor HvtA, that is needed in conjunction with the HupA protein for optimal hemin utilization. The HvtA protein is significantly homologous to other outer membrane hemin receptors and ...201122015545
identification of virulence factors in vibrio vulnificus by comparative transcriptomic analyses between clinical and environmental isolates using cdna microarray.we compared the gene expression among four clinical and five environmental v. vulnificus isolates, using a cdna microarray containing 131 genes possibly associated with pathogenicity, transport, signal transduction, and gene regulations in the pathogen. cdnas from total rnas of these isolates were hybridized into the cdna microarray using the cdna of the wild-type strain mo6/24-o as a reference. we focused on selecting differentially expressed (de) genes between clinical and environmental isolat ...201122210607
[septic shock by vibrio vulnificus at the coast gulf of mexico].vibrio vulnificus is a gram-negative bacterium which is found in marine environments and where there is a partly enclosed coastal body of water with one or more rivers or streams flowing into it, and with a free connection to the open sea. the infection by these bacteria can cause primary septicemia by two mechanisms: upon consuming crustaceans, mollusks and some fish (filtering shellfish) raw or barely cooked or by an open injury in contact with seawater. the patients with infections of the pr ...201121982195
[preparation and identification of ompw monoclonal antibodies].to prepare and characterize the mouse monoclonal antibodies against vibrio parahaemolyticus ompw.201121906472
high numbers of vibrio vulnificus in tar balls collected from oiled areas of the north-central gulf of mexico following the 2010 bp deepwater horizon oil spill.the deepwater horizon oil spill was the largest oil spill in usa history releasing approximately 4.9 million barrels of crude oil into the gulf of mexico. soon after the spill started, tar balls and other forms of weathered oil appeared in large numbers on beaches in mississippi and alabama. in this study, we analyzed tar balls for total aerobic bacterial (tab) counts and also for the presence of vibrio vulnificus, a human pathogen known to be abundant in the gulf coast environment and capable o ...201122109669
cooperative regulation of the vibrio vulnificus nan gene cluster by nanr protein, camp receptor protein, and n-acetylmannosamine 6-phosphate.the nan cluster of vibrio vulnificus, a food-borne pathogen, consists of two divergently transcribed operons, nant(psl)ar and nanek naga, required for transport and catabolism of n-acetylneuraminic acid (neu5ac). a mutation of nanr abolished the extensive lag phase observed for the bacteria growing on neu5ac and increased transcription of nant(p) and nane, suggesting that nanr is a transcriptional repressor of both nan operons. intracellular accumulation of neu5ac was dependent on the carbon sou ...201121956110
ß-1,3-Glucan recognition protein (ßGRP) is essential for resistance against fungal pathogen and opportunistic pathogenic gut bacteria in Locusta migratoria manilensis.Pattern recognition proteins, which form part of the innate immune system, initiate host defense reactions in response to pathogen surface molecules. The pattern recognition protein ß-1,3-glucan recognition protein (ßGRP) binds to ß-1,3-glucan on fungal surfaces to mediate melanization via the prophenoloxidase (PPO)-activating cascade. In this study, cDNA encoding a 53-kDa ßGRP (LmßGRP) was cloned from Locusta migratoria manilensis. LmßGRP mRNA shown to be constitutively expressed specifically i ...201122062247
relationship between localization on cellular membranes and cytotoxicity of vibrio vulnificus hemolysin.vibrio vulnificus secretes a hemolysin/cytolysin (vvh) that induces cytolysis in target cells. a detergent resistant membrane domain (drm) fraction of the cells after sucrose gradient centrifugation includes cholesterol-rich membrane microdomains which have been called "lipid rafts". it was reported that some pore-forming toxins require association with drm and/or lipid rafts to exert their cytotoxicity. it has also been thought that cellular cholesterol is involved in vvh cytotoxicity because v ...201122028805
aeromonas hydrophila sepsis mimicking vibrio vulnificus infection.aeromonas hydrophila is a facultatively anaerobic, asporogenous gram-negative rod that has often been regarded as an opportunistic pathogen in hosts with impairment of a local or general defense mechanism. a 68-year-old alcoholic woman presented with shock and gangrene on the right arm. at first, her clinical presentations were severe painful erythematous swelling that worsened within a few hours with development of gangrene, edema, and blisters. bullous fluid and blood cultures yielded a. hydro ...201122028565
Apoptotic effect of Vibrio vulnificus cytolysin on A549 human lung adenocarcinoma cells.Vibrio vulnificus cytolysin (VVC) has a very strong cytotoxic effect on various types of mammalian cells. However, the inhibitory effect of VVC on the proliferation of human lung cancer cells has scarcely been reported. This study aimed to analyze the effects of recombinant VVC (rVVC) on the A549 human lung adenocarcinoma cell line and to investigate the underlying molecular mechanisms governing these effects. This study showed that rVV ...201122134407
Use of a DNA microarray for detection and identification of bacterial pathogens associated with fishery products.We established a microarray for the simultaneous detection and identification of diverse putative pathogens often associated with fishery products by targeting specific genes of Listeria monocytogenes, Salmonella, Shigella, Staphylococcus aureus, Streptococcus pyogenes, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, and Yersinia enterocolitica and the 16S-23S rRNA gene internal transcribed spacer (ITS) region of Proteus mirabilis and Proteus vulgaris. The microarray contained 26 sp ...201121965411
Vibrio vulnificus IlpA induces MAPK-mediated cytokine production via TLR1/2 activation in THP-1 cells, a human monocytic cell line.Vibrio vulnificus is a pathogenic bacterium causing primary septicemia, which is followed by a classical septic shock pathway including an overwhelming inflammatory cytokine response. V. vulnificus IlpA is a potent immunogenic lipoprotein that triggers cytokine production in human monocytes by activating the toll-like receptor 2 (TLR2). In this study, we further defined the IlpA signaling pathways involved in cytokine production in the human monocytic cell line, THP-1. TLR2 was involved in cytok ...201121903273
Vaccination of sex reversed hybrid tilapia (Oreochromis niloticus × O. aureus) with an inactivated Vibrio vulnificus vaccine.Vibrio vulnificus causes disease in economically important aquaculture raised fish and is an opportunistic human pathogen. This study reports on the isolation of V. vulnificus from diseased hybrid tilapia (Oreochromis niloticus × O. aureus) cultured in a North American water reuse facility. Our objectives were to characterize the isolate using biochemical and molecular methods, develop a disease challenge model, and determine the ability of a formalin inactivated whole-cell vaccine to protect ag ...201121982852
[The effects of NF-E2-related factor-2 prompter polymorphism on alcoholic liver disease with Vibrio vulnificus sepsis].To investigate the influence of genetic polymorphism in NF-E2-related factor-2 (nrf2) gene promoter locus at 336 in alcoholic liver disease (ALD) with Vibrio vulnificus (VV) sepsis.201122169690
Vibrio vulnificus Diarrhea in a Child with Respiratory Infection.Vibrio vulnificus is a rare cause of disease and it is often unrecognized and underreported. It is a lactose-fermenting, halophilic vibrio causing severe disease in immunocompromised patients, but causing a mild type of gastroenteritis in healthy people, usually associated with consumption of seafood. We report here a case of diarrhea due to V. vulnificus in a male child who was admitted for fever, loose motions and productive cough. There was no history of consumption of any seafood; so, the so ...201121887066
Antibiotic therapy for necrotizing fasciitis caused by Vibrio vulnificus: retrospective analysis of an 8 year period.OBJECTIVES: To compare the effectiveness of a third-generation cephalosporin alone, a third-generation cephalosporin plus minocycline, and a fluoroquinolone in patients with necrotizing fasciitis (NF) caused by Vibrio vulnificus. METHODS: A retrospective review of case notes was performed for 89 patients who presented with NF caused by V. vulnificus and underwent surgical intervention within 24 h of admission between 2003 and 2010. Data on comorbidities, clinical manifestations, laboratory studi ...201122117030
protective mechanism of curcumin against vibrio vulnificus infection.curcumin, a natural polyphenolic flavonoid extracted from the rhizome of curcuma longa l., has many beneficial biological activities. however, there are relatively few reports of the effects of curcumin on pathogen infections. this study examined the effect of curcumin on a vibrio vulnificus infection. the cytotoxicity of v. vulnificus to hela cells was significantly inhibited by curcumin (at 10 or 30 μm). to further examine the inhibitory mechanism of curcumin against v. vulnificus-mediated cyt ...201122092562
In vitro evaluation of antibacterial activity of Asparagopsis taxiformis from the Straits of Messina against pathogens relevant in aquaculture.Ethanol extracts of Asparagopsis taxiformis collected from the Straits of Messina (Italy) were screened for antibacterial activity against pathogenic shellfish and fish bacteria previously isolated from local marine and brackish environments. Genetic labelling by DNA barcoding allowed us to identify the algal population as a biogeographical strain conspecific to A. taxiformis. The extract obtained in May showed the broadest antibacterial activity against all tested pathogenic bacteria, especiall ...201222078751
mechanistic insight into the catalytic activity of ββα-metallonucleases from computer simulations: vibrio vulnificus periplasmic nuclease as a test case.using information from wild-type and mutant vibrio vulnificus nuclease (vvn) and i-ppoi homing endonuclease co-crystallized with different oligodeoxynucleotides, we have built the complex of vvn with a dna octamer and carried out a series of simulations to dissect the catalytic mechanism of this metallonuclease in a stepwise fashion. the distinct roles played in the reaction by individual active site residues, the metal cation and water molecules have been clarified by using a combination of cla ...201122114054
Homodimerization and binding of specific domains to the target DNA are essential requirements for HlyU to regulate expression of the virulence gene rtxA1, encoding the repeat-in-toxin protein in the human pathogen Vibrio vulnificus.The virulence gene rtxA1, encoding the repeat-in-toxin protein, plays an essential role in the pathogenesis of Vibrio vulnificus infections. Expression of this gene is controlled by the HlyU regulator by direct contact of the DNA upstream of the rtxA1 toxin operon acting as a derepressor of the H-NS protein. The crystal structure suggests that HlyU forms a homodimer in vitro. However, knowledge of the biological implications of these findings in vivo is limited. In this work, we endeavored to di ...201122020641
year round patchiness of vibrio vulnificus within a temperate texas bay.aims:  to investigate with high geographical resolution the small-scale spatial and temporal distribution of the pathogen vibrio vulnificus throughout the water column in a temperate texas bay where numerous v. vulnificus infections had been reported by the regional media the previous summer. methods and results:  surface and bottom water samples were collected from 19 sites between april 2005 and october 2006 from matagorda bay, tx. physicochemical parameters were measured and v. vulnificus wer ...201122212214
pilf polymorphism-based real-time pcr to distinguish vibrio vulnificus strains of human health relevance.the gram-negative bacterium vibrio vulnificus is a common inhabitant of estuarine environments. globally, v. vulnificus is a significant foodborne pathogen capable of causing necrotizing wound infections and primary septicemia, and is a leading cause of seafood-related mortality. unfortunately, molecular methods for the detection and enumeration of pathogenic v. vulnificus are hampered by the genetically diverse nature of this pathogen, the range of different biotypes capable of infecting humans ...201222265278
comparison of molecular detection methods for vibrio parahaemolyticus and vibrio vulnificus.pathogenic vibrios are a global concern for seafood safety and many molecular methods have been developed for their detection. this study compares several molecular methods for detection of total and pathogenic vibrio parahaemolyticus and vibrio vulnificus, in mpn enrichments from oysters and fish intestine samples. this study employed the dupont qualicon bax(®) system real-time pcr assay for detection of v. parahaemolyticus and v. vulnificus. multiplex real-time pcr detection of total (tlh+), t ...201222265290
an extracellular serine protease produced by vibrio vulnificus ncimb 2137, a metalloprotease-gene negative strain isolated from a diseased eel.vibrio vulnificus is a ubiquitous estuarine microorganism but causes fatal systemic infections in immunocompromised humans, cultured eels or shrimps. an extracellular metalloprotease vvp/vvpe has been reported to be a potential virulence factor of the bacterium; however, a few strains isolated from a diseased eel or shrimp were recently found to produce a serine protease termed vvsa, but not vvp/vvpe. in the present study, we found that these strains had lost the 80 kb genomic region including t ...201122805945
antimicrobial activity of rice bran extracts for diarrheal disease.rice bran showed antioxidative, antimutagenic, carcinogenic and antibacterial activities in previous reports. the rice bran has been recently used as a natural source of health food for several diseases such as diabetes, atherosclerosis and cancer. severe diarrheal disease due to food-borne contamination of bacteria resulted from the bacteria have become resistant to many antibiotics. hence, early treatment of diarrhea using natural food containing antibacterial activity to prevent progression o ...201122619917
temporal and spatial variability in the distribution of vibrio vulnificus in the chesapeake bay: a hindcast study.vibrio vulnificus, an estuarine bacterium, is the causative agent of seafood-related gastroenteritis, primary septicemia, and wound infections worldwide. it occurs as part of the normal microflora of coastal marine environments and can be isolated from water, sediment, and oysters. hindcast prediction was undertaken to determine spatial and temporal variability in the likelihood of occurrence of v. vulnificus in surface waters of the chesapeake bay. hindcast predictions were achieved by forcing ...201122302219
genetic diversity of the human pathogen vibrio vulnificus: a new phylogroup.the biotype 3 group of the human pathogen vibrio vulnificus emerged in israel probably as a result of genome hybridization of two bacterial populations. we performed a genomic and phylogenetic study of v. vulnificus strains isolated from the environmental niche from which this group emerged - fish aquaculture in israel. the genetic relationships and evolutionary aspects of 188 environmental and clinical isolates of the bacterium were studied by genomic typing. genetic relations were determined b ...201122227412
genetic and quantitative assessment of vibrio vulnificus populations in oyster (crassostrea virginica) tissues.vibrio vulnificus is a leading cause of shellfish-associated food-borne illness. us regulations stipulate shellfish processing procedures to limit v. vulnificus densities; however, the effect of these procedures on v. vulnificus strain distribution and/or genetic diversity is unknown. vibrio vulnificus concentrations and strain diversity were analysed in various oyster tissues stored overnight at 26°c that were subsequently divided into two treatment groups: one received post-harvest processing ...201123761333
effective reduction of vibrio vulnificus in the eastern oyster (crassostrea virginica) using high salinity depuration.depuration under different salinities was used to reduce the human pathogen vibrio vulnificus from eastern oysters (crassostrea virginica). individual recirculating systems were used to test the efficacy of depuration at three salinities (15, 25, and 35 psu) in four independent trials during a 14 day period. initial loads of v. vulnificus were higher than 10,000 mpn/g of oyster meat in all trials. data showed that 25 and 35 psu treatments were more efficient in reducing v. vulnificus numbers tha ...201223498187
complete genome sequence of bacteriophage vvaw1, which infects vibrio vulnificus.investigating the bacteriophages of vibrios has led to significant insights into the evolution and pathogenicity of their host strains. this report presents the first complete genome sequence of a bacteriophage that infects the deadly human pathogen vibrio vulnificus. the phage was isolated from the surface waters of the ala wai canal, which is part of an urban watershed in eastern honolulu, hawaii, usa. the phage particle is icosahedral, with a diameter of 35-40 nm, and a small non-contractile ...201223408718
studies on water quality and pathogenic bacteria in coastal water langkawi, malaysia.a study on physico-chemical parameters and pathogenic bacterial community was carried out at the coastal waters of pulau tuba island, langkawi. the physico-chemical parameters such as temperature (27.43-28.88 degrees c), dissolved oxygen (3.79-6.49 mg l(-1)), ph (7.72-8.20), salinity (33.10-33.96 ppt), total dissolved solids (32.27-32.77 g l(-1)) and specific conductivity (49.83-51.63 ms cm(-1)) were observed. station 3 and station 4 showed highest amount of nitrates (26.93 and 14.61 microg at n ...201223360015
effect of temperature on growth of vibrio paraphemolyticus and vibrio vulnificus in flounder, salmon sashimi and oyster meat.vibrio parahaemolyticus and vibrio vulnificus are the major pathogenic vibrio species which contaminate ready-to-eat seafood. the purpose of this study was to evaluate the risk of human illness resulting from consumption of ready-to-eat seafood such as sashimi and raw oyster meat due to the presence of v. parahaemolyticus and v. vulnificus. we compared the growth kinetics of v. parahaemolyticus and v. vulnificus strains in broth and ready-to-eat seafood, including flounder and salmon sashimi, as ...201223330227
integration of vibrio vulnificus into marine aggregates and its subsequent uptake by crassostrea virginica oysters.marine aggregates are naturally forming conglomerations of larvacean houses, phytoplankton, microbes, and inorganics adhered together by exocellular polymers. in this study, we show in vitro that the bacterial pathogen vibrio vulnificus can be concentrated into laboratory-generated aggregates from surrounding water. we further show that environmental (e-genotype) strains exhibit significantly more integration into these aggregates than clinical (c-genotype) strains. experiments where marine aggr ...201223263962
vibrio vulnificus: death on the half shell. a personal journey with the pathogen and its ecology.vibrio vulnificus is an estuarine bacterium which occurs in high numbers in filter-feeding molluscan shellfish, such as oysters. in individuals with certain underlying diseases, ingestion of the bacterium, e.g., in raw or undercooked oysters, can lead to a rapid and extremely fatal infection. indeed, this one bacterium is responsible for 95 % of all seafood-borne deaths. in addition, the bacterium is capable of entering a preexisting lesion or cut obtained during coastal recreational activities, ...201223263234
[primary transport of acute patients. optimal use of rescue resources exemplified by the canton of graubünden]. 20113927607
root caries. is it the caries problem of the future? 20143896444
inducibility of metallothionein biosynthesis in cultured normal and menkes kinky hair disease fibroblasts: effects of copper and cadmium.metallothionein biosynthesis is not induced by extracellular copper in menkes kinky hair disease (mkhd) or in normal cultured fibroblasts under the conditions of these experiments. in the presence of copper, mkhd fibroblasts also incorporated less cysteine than did normal fibroblasts. extracellular cadmium greatly enhanced the uptake of cysteine in both normal and mkhd cultures. by the technique of polyacrylamide gel electrophoresis, it was demonstrated that metallothionein is induced by cadmium ...2004471576
[conservative management of postmastectomy lymphedema]. 2004419468
[low-molecular weight nuclear rna's]. 2004395786
carcinoma of the male breast metastatic to the mandible.a case is reported of male breast carcinoma which, among other bony sites, metastasized to the mandible. this case and the review of the literature illustrate the need to include the mandible in bone scans for metastatic disease in breast cancer.20051310607
[practical evaluation of methods in pharmacokinetics].a short discussion of the differences between kinetic processes of zero and first order is followed by an outline of evaluation methods used in pharmacokinetic practice, e.g. "paper and pencil methods" as well as the use of digital and analog computers. the increasing potency of new drugs requires not only more sophisticated methods of analysis and expensive facilities but also more complex models to generate pharmacokinetic information.20051242344
[production of tropane alkaloids in tissue cultures (author's transl)]. 20051187871
failure to consult specialist. 20051173090
morphometric analysis of the loss of chief and parietal cells after partial gastrectomy for duodenal ulcer.relative chief and parietal cell volume densities were estimated morphometrically in the remnant mucosa of 98 male patients ("series"), operated on for duodenal ulcer by the billroth ii, and in the body mucosa of 55 subjects, age and sex matched, from a random series of a finnish population ("controls"). the relative volumes of chief and parietal cells were significantly lower in the series than in the controls. the mean chief cell: parietal cell ratio was significantly higher in the series than ...2005549432
acute myocardial infarction after blunt chest trauma in young people: need for prompt intervention.we describe four cases of acute myocardial infarction in young patients, secondary to blunt chest trauma. one case was treated with intracoronary thrombolysis and angioplasty, two cases received systemic thrombolysis, and the last one did not have any reperfusion therapy. the coronary angiograms of the left anterior descending artery showed thrombosis in two cases, coronary dissection in one case, and no morphological lesions in the other. we encourage the early performance of angiographic studi ...20061764738
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