Publications

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a simple and inexpensive method for preparing erythrocyte membranes by filtration through a hollow-fiber system.an efficient, simple method for preparing biologically active erythrocyte membranes is described. the semi-automated procedure involves circulating hemolysate mixture through a hollow-fiber system, thereby filtering off intracellular components leaving a high yield of washed ghosts. the prepared ghosts exhibit cation-stimulated atpase activities comparable to those of ghosts prepared by traditional methods. electron micrographs revealed that the filtration isolation caused less shearing of the m ...20082527009
[diet in the prevention of coronary disease]. 20083815423
[diet in the prevention of coronary disease]. 20083815423
[reiter's syndrome or lyme disease?]. 20082632003
[reiter's syndrome or lyme disease?]. 20082632003
hepatitis b surface antigen seroconversion in resident doctors. 20082292578
management of the temporomandibular joint surgical patient.comprehensive management of the temporomandibular joint surgical patient necessitates a thorough presurgical evaluation and definitive postsurgical management. the article provides an overview of tmj surgical techniques, including arthroscopy, which has significantly decreased the need for open joint procedures. early postsurgical intervention is recommended and specific therapeutic paradigms are outlined. adjunctive technique to reduce inflammation, muscle guarding, edema, and collagen bonding ...20082605859
american academy of pediatrics: prep statement. 20082587145
[prostheses on osteointegrated implants: restoration of a missing tooth (4)].this article is the last of a series of four. this article describes in detail the fabrication of a crown connected to a brånemark implant.20082394869
population structures of two genotypes of vibrio vulnificus in oysters (crassostrea virginica) and seawater.vibrio vulnificus biotype 1 strains can be classified into two genotypes based on the pcr analysis of variations in the virulence-correlated gene (vcg). genotype has been correlated with human infection for 90% of isolates from human cases having the vcgc sequence type and 87% of environmental strains having the vcge variant. in this study we examined the dynamics of v. vulnificus populations and the distribution of the two genotypes recovered from oysters and surrounding estuarine wasters. anal ...200817993556
phylogenetic analysis of the incidence of lux gene horizontal transfer in vibrionaceae.horizontal gene transfer (hgt) is thought to occur frequently in bacteria in nature and to play an important role in bacterial evolution, contributing to the formation of new species. to gain insight into the frequency of hgt in vibrionaceae and its possible impact on speciation, we assessed the incidence of interspecies transfer of the lux genes (luxcdabeg), which encode proteins involved in luminescence, a distinctive phenotype. three hundred three luminous strains, most of which were recently ...200818359809
studies on diseased freshwater prawn macrobrachium rosenbergii infected with vibrio vulnificus.the present study was aimed at isolation and characterization of the pathogenic bacterium from diseased freshwater prawn. the effect of the bacterial pathogen on hepatopancreas, gills and exoskeleton was also investigated. diseased freshwater prawn, macrobrachium rosenbergii were collected from commercial hatchery in behera governorate, egypt. the diseased prawn showed dark brown focal lesions and necrosis of appendage tips. the causative bacterial pathogen was isolated from haemolymph and hepat ...200819266922
[a protocol for diagnosis and treatment of vibrio vulnificus sepsis].vibrio vulnificus sepsis is one of the most fatal disease with a high mortality which exceeds 50%. but at present there is no evidence-based guidelines for diagnosis and therapy of vibrio vulnificus sepsis because of its dispersion in occurrence and low incidence.200818199391
vibrio vulnificus-induced death of jurkat t-cells requires activation of p38 mitogen-activated protein kinase by nadph oxidase-derived reactive oxygen species.vibrio vulnificus, a pathogenic bacterium causing primary septicemia, exhibited cytotoxicity towards jurkat cells of t-lymphocytes through intracellular reactive oxygen species (ros) production. pretreatment of jurkat t-cells with diphenyleneiodonium chloride (dpi) abolished v. vulnificus-induced ros generation and bacterial ability to cause cell death. jurkat t-cells expressing dominant-negative protein of rac subunit of nadph oxidase (nox) did not show increased ros production and cell death b ...200818571150
vibrio vulnificus rpos expression is repressed by direct binding of camp-camp receptor protein complex to its two promoter regions.vibrio vulnificus, a septicemia-causing pathogenic bacterium, acquires resistance against various stresses and expresses virulence factors via an rpos gene product. in this study, we investigated the transcriptional characteristics of this global regulator. two distinct transcriptional initiation sites for the rpos gene, the proximal promoter (p(p)) and the distal promoter (p(d)), were defined by primer extension experiments. various rpos::luxab transcriptional fusions indicated that p(d) is a m ...200818713737
differentiating between isolates of vibrio vulnificus with monoclonal antibodies.monoclonal antibodies (mabs) against vibrio vulnificus (isolate i, vvc and isolate ii, vvb) were raised using heat-killed and heat-killed plus sds-mercaptoethanol treated forms of vvc and vvb for immunizing swiss mice. twenty three hybridomas producing mabs against v. vulnificus were selected and divided into five groups according to their specificities to different v. vulnificus isolates and apparent protein antigens which ranged from approximately 3-50 kda. four groups were specific to v. vuln ...200818706941
immunomagnetic separation and coagglutination of vibrio parahaemolyticus with anti-flagellar protein monoclonal antibody.mice were immunized by injection of vibrio parahaemolyticus atcc 17802 polar flagellin in order to produce monoclonal antibodies (mabs). mabs were analyzed by anti-h enzyme-linked immunosorbent assay using v. parahaemolyticus polar flagellar cores. the mab exhibiting the highest anti-h titer was coated onto cowan i staphylococcus aureus cells at a concentration of 75 microg/ml cell suspension and used for slide coagglutination. of 41 isolates identified genetically as v. parahaemolyticus, 100% c ...200818753337
relative stability of helices determines the folding landscape of adenine riboswitch aptamers.riboswitches, whose folding is controlled by binding of metabolites to the aptamer domain, regulate downstream gene expression. folding properties of the aptamer strongly influence the conformation of the downstream expression platform, which controls transcription termination or translation initiation. we have characterized the energy landscape of the add riboswitch aptamer quantitatively by unfolding and refolding the molecule with mechanical force using the coarse-grained self-organized polym ...200818828635
regulation of fatty acid metabolism by fadr is essential for vibrio vulnificus to cause infection of mice.the opportunistic bacterial pathogen vibrio vulnificus causes severe wound infection and fatal septicemia. we used alkaline phosphatase insertion mutagenesis in a clinical isolate of v. vulnificus to find genes necessary for virulence, and we identified fadr, which encodes a regulator of fatty acid metabolism. the fadr::mini-tn5km2phoa mutant was highly attenuated in a subcutaneously inoculated iron dextran-treated mouse model of v. vulnificus disease, was hypersensitive to the fatty acid syntha ...200818835990
vibrio vulnificus rtx toxin plays an important role in the apoptotic death of human intestinal epithelial cells exposed to vibrio vulnificus.during vibrio vulnificus infection, v. vulnificus reaches the intestine and then invades the bloodstream by crossing the intestinal mucosal barrier of the host, which results in systemic septicemia. previously, we reported that the rtxa toxin secreted through the rtxe transporter contributes to the cytotoxicity of v. vulnificus against intestinal epithelial cells. here, we used gene mutants of rtxe and rtxa to determine the role that v. vulnificus rtxa toxin plays in the apoptotic death of human ...200818849006
clinical characteristics and molecular subtyping of vibrio vulnificus illnesses, israel. 200819046510
new approach for discrimination of vibrio vulnificus by real-time pcr before and after gamma-irradiation.the effects of gamma-irradiation on the destruction of vibrio vulnificus by real-time pcr were studied. gamma-irradiation was found to result in extensive reduction in the molecular size of dna. irradiation of viable cells (1 x 10(6) cfu/ml) at 1.08 kgy resulted in 100% destruction determined by plate counts, with most of the dna from the irradiated cells having a bp-length of less than 1000. the use of a pair of primers to amplify a 1000-bp sequence of dna from cells exposed to 1.08 kgy failed ...200818262296
further characterization of vibrio vulnificus rugose variants and identification of a capsular and rugose exopolysaccharide gene cluster.capsular polysaccharide (cps) is a major virulence factor in vibrio vulnificus, and encapsulated strains have an opaque, smooth (ops) colony morphology, while nonencapsulated strains have a translucent, smooth (trs) colony morphology. previously, we showed that ops and trs parental strains can yield a third colony type, rugose (r), and that the resulting strains, with the opr and trr phenotypes, respectively, form copious biofilms. here we show that while opr and trr strains both produce three-d ...200818212074
protective effect of polygoni cuspidati radix and emodin on vibrio vulnificus cytotoxicity and infection.vibrio vulnificus, a good model organism of bacterial septicemia, causes fatal septicemia manifesting a fulminating course and a high mortality rate within days. in order to identify new natural substances preventing v. vulnificus infection, a plant library was screened for inhibiting cytotoxicity to host cells by using trypan blue staining and ldh assay. we found that polygoni cuspidati radix potently suppressed the acute death of hela and raw264.7 cells in a dose dependent manner. further stud ...200819107405
the extracellular metalloprotease of vibrio tubiashii is a major virulence factor for pacific oyster (crassostrea gigas) larvae.vibrio tubiashii is a recently reemerging pathogen of larval bivalve mollusks, causing both toxigenic and invasive disease. marine vibrio spp. produce an array of extracellular products as potential pathogenicity factors. culture supernatants of v. tubiashii have been shown to be toxic to oyster larvae and were reported to contain a metalloprotease and a cytolysin/hemolysin. however, the structural genes responsible for these proteins have yet to be identified, and it is uncertain which extracel ...200818456850
evaluation of a loop-mediated isothermal amplification assay for detecting vibrio vulnificus in raw oysters.human consumption of raw or undercooked seafood, particularly oysters, may lead to severe infections due to the presence of vibrio vulnificus. in this study, a sensitive and specific loop-mediated isothermal amplification (lamp) assay was developed to detect this pathogen in raw oysters. two outer and two inner primers were designed to specifically recognize the v. vulnificus cytolysin/hemolysin gene (vvha), and the reaction could be completed in 1 hour at 63 degrees c. direct visual observation ...200818564911
[changes of toll-like receptor, tnf-alpha and il-10 in liver tissue of rats before and after intragastric infusion with alcohol with vibrio vulnificus sepsis].to detect the effects of antimicrobial agents on the toll-like receptor (tlr) and so on in liver tissue of rats after intragastric infusion with alcohol with vibrio vulnificus (vv) sepsis.200819175176
deferasirox plus ciprofloxacin combination therapy after rapid diagnosis of vibrio vulnificus sepsis using real-time polymerase chain reaction.we report a case of vibrio vulnificus sepsis developed during deferoxamine therapy for transfusional iron overload due to secondary hemochromatosis of myelodysplastic syndrome. the patient was treated with adjuvant deferasirox in combination with ciprofloxacin, after rapid diagnosis of v. vulnificus sepsis using real-time polymerase chain reaction (pcr). this case suggests that since v. vulnificus dna can be detected by real-time pcr for several days even after patients receive antibiotics, real ...200818930321
nonfoodborne vibrio infections: an important cause of morbidity and mortality in the united states, 1997-2006.infections due to vibrio species cause an estimated 8000 illnesses annually, often through consumption of undercooked seafood. like foodborne vibrio infections, nonfoodborne vibrio infections (nfvi) also result in serious illness, but awareness of these infections is limited.200818444811
comparison of conventional, nested, and real-time pcr assays for rapid and accurate detection of vibrio vulnificus.we conducted a prospective study to target toxr in the blood of patients with skin and soft tissue infections who were admitted to four tertiary hospitals to assess the clinical usefulness of real-time quantitative pcr (q-pcr) as a diagnostic technique. we performed conventional pcr (c-pcr), nested pcr (n-pcr), and q-pcr assays and compared the results to those obtained using the "gold standard" of microbiological culture. the lower detection limit for the q-pcr assay was 5 x 10(0) copies/microl ...200818614647
[taxonomic characteristics and physiological properties of microorganisms from the gut of pike (esox lucius)].the taxonomic composition and distribution of microorganisms differing in the degree of association with the intestinal mucosa of the pike (lucius lucius) has been studied. microorgansism of the families enterobacteriaceae, aeromonadaceae, and vibrionaceae dominate in the gut microflora. numerically prevailing bacterial species are characterized by high proteolytic and amylolytic enzyme activities as well as by high persistence accounted for by antilysozyme and antihistone activities. the result ...200819198074
[rapid detection of vibrio vulnificus using a loop-mediated isothermal amplification method].vibrio vulnificus is found in marine waters near the coast around the world. infection with this gram-negative rod, via ingestion of raw seafood or via a skin wound following contact with contaminated estuarine or marine water, can cause necrotizing fasciitis and sepsis. most of patients with vibrio vulnificus infection have underlying liver dysfunction or diabetes mellitus. due to the high mortality and short latent periods, control of this infection depends on early identification of the bacte ...200818975582
application of the rpos gene for species-specific detection of vibrio vulnificus by real-time pcr.vibrio vulnificus is a causative agent of serious diseases in humans, resulting from the contact of wound with seawater or consumption of raw seafood. several studies aimed at detecting v. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. in this study, we targeted the rpos gene, a general stress regulator, to detect v. vulnificus. pcr specificity was identified by amplification of 8 v. vulnificus templates and by the loss of a pcr product with 36 ...200819047830
microbial and histopathological study of the vibriosis caused by vibrio vulnificus serovar e in eels: the metalloprotease vvp is not an essential lesional factor.vibrio vulnificus biotype 2 serovar e (bt2-sere) is a zoonotic pathogen that causes a haemorrhagic septicaemia in eels, called warm water vibriosis. the main objective of the present work was to study the onset of the eel vibriosis from the microbiological and histopathological viewpoint, as well as to ascertain the role of the protease vvp as a lesional factor by comparing the histopathological lesions caused by the wild strain and its vvp deficient derivative. the wild-type strain was observed ...200818930131
structure-function relationships in escherichia coli adenylate cyclase.class i adenylate cyclases are found in gamma- and delta-proteobacteria. they play central roles in processes such as catabolite repression in escherichia coli or development of full virulence in pathogens such as yersinia enterocolitica and vibrio vulnificus. the catalytic domain (residues 2-446) of the adenylate cyclase of e. coli was overexpressed and purified. it displayed a v(max) of 665 nmol of camp x mg(-1) x min(-1) and a k(m) of 270 microm. titration of the metal cofactor mg(2+) against ...200818620542
molecular modeling and docking studies of glutamate racemase in vibrio vulnificus cmcp6.identification of novel drug targets in silico in vibrio vulnificus is important as it is one of the emerging pathogenic microorganisms. glutamate racemase, an important constituent of bacterial cell wall is chosen for structure prediction using homology modeling. with the aid of tools and software like modeller and swiss-pdbviewer, the 3d structure is predicted and the final model is refined by energy minimization. the quality of the refined model is assessed using procheck. the interaction bet ...200819374132
diagnosing and treating deadly vibrio vulnificus infection. 200818358354
development of quantitative real-time polymerase chain reaction for the detection of vibrio vulnificus based on hemolysin (vvha) coding system.to establish a taqman real-time fluorescent quantitative pcr to detect vibrio vulnificus based on the hemolysin gene (vvha) coding cytolysin.200818837292
vibrio vulnificus biotype 2 serovar e gne but not gale is essential for lipopolysaccharide biosynthesis and virulence.this work aimed to establish the role of gne (encoding udp-galnac 4-epimerase activity) and gale (encoding udp-gal-4-epimerase activity) in the biosynthesis of surface polysaccharides, as well as in the virulence for eels and humans of the zoonotic serovar of vibrio vulnificus biotype 2, serovar e. dna sequence data revealed that gne and gale are quite homologous within this species (> or =90% homology). mutation in gne of strain cect4999 increased the surface hydrophobicity, produced deep alter ...200818227162
variable repeat regions in the genome of vibrio vulnificus and polymorphism in one of the loci in strains isolated from oysters.vibrio vulnificus an estuarine bacterium is associated with severe wound infections and fatal septicemia related to consumption of raw shellfish. in this study we screened the two whole genome sequences available for v. vulnificus in genbank for the presence of variable number of tandem repeat (vntr) regions. five potential vntr loci with unit repeat size ranging from 6-7 nucleotides were identified for v. vulnificus genome. one of the loci designated vv1 was selected to detect the repeat number ...200818374439
discrimination of gamma-irradiated and nonirradiated vibrio vulnificus by using real-time polymerase chain reaction.to develop a pcr strategy for vibrio vulnificus in irradiated foods.200818081775
crystallization and preliminary x-ray diffraction analysis of the hsdr subunit of a putative type i restriction enzyme from vibrio vulnificus yj016.type i restriction enzymes are multimeric proteins that consist of three subunits. the hsds and hsdm subunits form a complex protein that shows methyltransferase activity, while the hsdr subunit functions as an endonuclease as well as as a translocase. of these three subunits, no structural information on the hsdr subunit is yet available. the putative hsdr gene from vibrio vulnificus yj016 (hsdr_vv) was cloned and expressed and the expressed protein hsdr_vv was purified. hsdr_vv was crystallize ...200818931437
efficiency of real-time polymerase chain reaction assay to detect vibrio vulnificus in seawater.the growth of vibrio vulnificus in an enriched culture of seawater during the summer in japan was monitored by a plating technique used as the culture method and a real-time polymerase chain reaction (pcr) assay as the molecular method. v. vulnificus was detected by the real-time pcr assay in the samples of august and september but not by the culture method. vibrio parahaemolyticus, however, was detected among all of the samples with both the culture method and real-time pcr assay. in the analys ...200818162438
effectiveness of icing as a postharvest treatment for control of vibrio vulnificus and vibrio parahaemolyticus in the eastern oyster (crassostrea virginica).the focus of this research was to investigate the efficacy of icing as a postharvest treatment for reduction of the levels of vibrio vulnificus and vibrio parahaemolyticus in commercial quantities of shellstock oysters. the experiments were conducted in june and august of 2006 and consisted of the following treatments: (i) on-board icing immediately after harvest; (ii) dockside icing approximately 1 to 2 h prior to shipment; and (iii) no icing (control). changes in the levels of pathogenic vibri ...200818680950
multiplex pcr assay for detection and simultaneous differentiation of genotypes of vibrio vulnificus biotype 1.a multiplex pcr assay was developed that provides a rapid method of species identification with simultaneous genotype differentiation of vibrio vulnificus biotype 1. this technique has been employed for the confirmation and typing of unknown isolates from both clinical and environmental sources utilizing a single reaction followed by gel electrophoresis.200818687035
detection of quorum-sensing-related molecules in vibrio scophthalmi.cell-to-cell communication (also referred to as quorum sensing) based on n-acyl-homoserine lactones (ahls) is a widespread response to environmental change in gram-negative bacteria. ahls seem to be highly variable, both in terms of the acyl chain length and in the chemical structure of the radicals. another quorum sensing pathway, the autoinducer-2-based system, is present both in gram-positive and gram-negative bacteria. in this study the presence of signal molecules belonging to both quorum s ...200818700048
nonribosomal peptide synthase is responsible for the biosynthesis of siderophore in vibrio vulnificus mo6-24/o.vibrio vulnificus produces siderophores, lowmolecular- weight iron-chelating compounds, to obtain iron under conditions of iron deprivation. to identify genes associated with the biosynthesis of siderophore in v. vulnificus mo6-24/ o, we screened clones with mini-tn5 random insertions for those showing decreased production of siderophore. among 6,000 clones screened, nine such clones were selected. these clones contain the transposon inserted in vv2_0830 (genbank accession number) that is a homo ...200818239413
real-time pcr assays for quantification and differentiation of vibrio vulnificus strains in oysters and water.vibrio vulnificus is an autochthonous estuarine bacterium and a pathogen that is frequently transmitted via raw shellfish. septicemia can occur within 24 h; however, isolation and confirmation from water and oysters require days. real-time pcr assays were developed to detect and differentiate two 16s rrna variants, types a and b, which were previously associated with environmental sources and clinical fatalities, respectively. both assays could detect 10(2) to 10(3) v. vulnificus total cells in ...200818245234
identification of lacto-n-biose i phosphorylase from vibrio vulnificus cmcp6.a beta-1,3-galactosyl-n-acetylhexosamine phosphorylase (galglynacp) homolog gene was cloned from vibrio vulnificus cmcp6. in synthetic reactions, the recombinant enzyme acted only with glcnac and galnac as acceptors in the presence of alpha-d-galactose-1-phosphate as a donor to form lacto-n-biose i (lnb) (galbeta1 --> 3glcnac) and galacto-n-biose (gnb) (galbeta1 --> 3galnac), respectively. glcnac was a much better acceptor than galnac. the enzyme also phosphorolysed lnb faster than it phosphorol ...200818723650
epidemiological survey of vibrio vulnificus infection in japan between 1999 and 2003.the frequency of vibrio vulnificus infection is very rare and there are many questions regarding its epidemiology in japan. to investigate the clinical course and epidemiology of v. vulnificus infection in japan, we performed a retrospective questionnaire survey in which 1693 hospitals from all over japan were surveyed, including advanced life saving emergency centers and dermatology institutions. of the 1693 hospitals, we received answers from 1045. ninety-four cases were confirmed as v. vulnif ...200818346255
bacteria flora and heavy metals in cultivated oysters crassostrea iredalei of setiu wetland, east coast peninsular malaysia.slipper oyster crassostrea iredalei is a species of good demand for its sweet flavor and white coloured flesh. the filter feeding nature predisposes oysters to accumulation of pathogenic and heavy metals in waters impacted by sewage pollutions and may thus render the oysters unfit for human consumption. a study was undertaken to investigate the presence of bacteria flora and heavy metal concentrations in cultivated oysters crassostrea iredalei at setiu wetland, terengganu, the only source of cul ...200818369732
sudden death caused by fulminant bacterial infection: background and pathogenesis of japanese adult cases.to analyze a risk factor for the onset of fulminant bacterial infection.200818758124
surveillance for waterborne disease and outbreaks associated with recreational water use and other aquatic facility-associated health events--united states, 2005-2006.since 1971, cdc, the u.s. environmental protection agency, and the council of state and territorial epidemiologists have collaboratively maintained the waterborne disease and outbreak surveillance system for collecting and reporting data related to waterborne-disease outbreaks (wbdos) associated with drinking water. in 1978, wbdos associated with recreational water (natural and treated water) were added. this system is the primary source of data regarding the scope and effects of disease associa ...200818784642
surveillance of health status on eight marine rainbow trout, oncorhynchus mykiss (walbaum), farms in denmark in 2006.the health status of eight marine rainbow trout farms was followed from mid-june to mid-september 2006 by sampling both dead and healthy fish approximately every 2 weeks for bacteriological and virological investigation. no fish pathogenic viruses were detected, but all farms experienced disease and mortality as a result of various bacterial infections. yersinia ruckeri was found on four and renibacterium salmoninarum on five of the farms, but only during the first part of the surveillance perio ...200818786028
analysis of gene expression in mouse alveolar macrophages stimulated with quorum-sensing mutants of vibrio vulnificus.bacterial pathogens manipulate host cells to promote pathogen survival and dissemination. in this study, microarray technology was used to identify the genes that are affected by the vibrio vulnificus quorum-sensing genes, luxs and smcr. by comparing the expression profiles of mouse macrophage cell lines stimulated with either the parent strains or a luxs smcr mutant, differentially expressed genes were identified. the genes included those that affect host cell death, stress, signaling transduct ...200818806354
csra inhibits the formation of biofilms by vibrio vulnificus.pcr screening of the shellfish-borne pathogen vibrio vulnificus revealed csra-negative strains, and these strains formed increased biofilm compared to csra-positive strains. complementation in trans with csra resulted in reduced biofilm formation, similar to that by csra(+) strains. our results provide evidence that csra inhibits biofilm formation in v. vulnificus.200818820071
occurrence, seasonality and genetic diversity of vibrio vulnificus in coastal seaweeds and water along the kii channel, japan.vibrio vulnificus is a ubiquitous toxigenic bacterium found in a coastal environment but little is known about its occurrence and seasonality among seaweeds, which are widely consumed as seafood in japan. therefore, we have observed the bacterium's abundance in seawater and seaweed samples from three areas of the kii channel, japan, during june 2003 to may 2004. a total of 192 samples were collected: 24 from each source in summer, autumn, winter and spring. the samples were selectively cultivate ...200818355295
two forms of vibrio vulnificus metalloprotease vvpe are secreted via the type ii general secretion system.vibrio vulnificus has been known to secrete one form of metalloprotease vvpe (45 kda) that is cleaved to 34 kda-vvpe and 11 kda-c-terminal propeptide via extracellular autoproteolysis. however, we found that extracellular secretion of both the 34 and 45 kda forms of vvpe began in the early growth phase; moreover, 34 kda-vvpe existed as the major form in v. vulnificus cell lysates and culture supernatants. in addition, extracellular secretion of both 34 and 45 kda-vvpe was blocked by mutation of ...200818604505
detection and characterization of a functional insertion sequence, isvpa2, in vibrio parahaemolyticus.pcr analysis of the pandemic strain of vibrio parahaemolyticus, kx-v237 (total genome sequenced) showed a subculture where the size of the amplicons had increased. the purpose of this study was to analyze the mechanism of this change. we found a 1,243-bp dna sequence inserted in one of the pandemic marker genes in this strain. the inserted dna sequence possessed the genetic structures shared by insertion sequences (iss) of the is3 family. this is had 26-bp imperfect terminal inverted repeats (ir ...200818164873
conditions for a 5-log reduction of vibrio vulnificus in oysters through high hydrostatic pressure treatment.vibrio vulnificus is frequently associated with oysters, and since oysters are typically consumed raw on a half shell, they can pose a threat to public health due to ingestion of this pathogenic marine microorganism. oysters should be processed to reduce the number of this pathogen. high pressure processing is gaining more and more acceptance among oyster processors due to its ability to shuck oysters while keeping the fresh-like characteristics of oysters. nine strains of v. vulnificus were tes ...200818177963
a case of vibrio vulnificus keratitis. 200818626738
proteomic identification and characterization of vibrio vulnificus proteins induced upon exposure to int-407 intestinal epithelial cells.proteomic analysis led to identification of the proteins of vibrio vulnificus that were induced upon exposure to int-407 cells, and 7 of which belong to the functional categories such as amino acid transport/metabolism, nucleotide transport/metabolism, posttranslational modification/protein turnover/chaperones, and translation. among the genes encoding the host-induced proteins, disruption of purh, trpd, tsaa, and groel2 resulted in reduced cytotoxicity. the purh, trpd, and tsaa mutants showed i ...200818633300
vibrio vulnificus infection presenting with compartment syndrome of bilateral calves. 200818572348
global gene expression as a function of the iron status of the bacterial cell: influence of differentially expressed genes in the virulence of the human pathogen vibrio vulnificus.vibrio vulnificus multiplies rapidly in host tissues under iron-overloaded conditions. to understand the effects of iron in the physiology of this pathogen, we performed a genome-wide transcriptional analysis of v. vulnificus growing at three different iron concentrations, i.e., iron-limiting [trypticase soy broth with 1.5% nacl (tsbs) plus ethylenediamine-di-(o-hydroxyphenylacetic) acid (edda)], low-iron (1 microg fe/ml; tsbs), and iron-rich (38 microg fe/ml; tsbs plus ferric ammonium citrate) ...200818573903
a consensus sequence for binding of smcr, a vibrio vulnificus luxr homologue, and genome-wide identification of the smcr regulon.quorum sensing has been implicated as an important global regulatory system controlling the expression of numerous virulence factors in bacterial pathogens. in the present study, dna targets of smcr, a vibrio vulnificus luxr homologue, were selected from a random pool of dna fragments by using a cycle selection procedure consisting of in vitro dna-smcr interaction, purification of smcr-dna complexes, and pcr amplification of smcr-bound dna. the amplified dna fragments were cloned and analyzed se ...200818579523
lower extremity infections by vibrio vulnificus.vibrio vulnificus is a gram-negative pathogen which is found in seawater and shellfish during warm months and can cause local infections in healthy individuals or septicemia in patients with chronic liver disease. materials-methods: clinical and laboratory records of four complicated cases are presented, with a 4.2 mean year follow-up.200818457099
achilles tendinitis in vibrio vulnificus sepsis: a rare presentation.vibrio vulnificus infection commonly presents as sepsis or wound infection. a case of alcoholic liver cirrhosis developed acute left heel pain and swelling 2 days after ingesting raw fish. two sets of blood cultures indicated v. vulnificus. hla-b27 was negative. ultrasonography revealed left achilles tendinitis. fluid aspirated from the left retrocalcaneal bursa was sterile. the patient recovered completely 3 weeks after treatment. to our knowledge, v. vulnificus septicaemia with achilles tendin ...200818465460
inducible resistance of fish bacterial pathogens to the antimicrobial peptide cecropin b.cecropin b is a cationic antimicrobial peptide originally isolated from the diapausing pupae of the giant silk moth, hylphora cecropia. cecropin b elicits its antimicrobial effects through disruption of the anionic cell membranes of gram-negative bacteria. previous work by our laboratory demonstrated that a constitutively expressed cecropin b transgene conferred enhanced resistance to bacterial infection in medaka. the development of antibiotic resistance by pathogenic bacteria is a growing prob ...200818474580
cyclic-di-gmp regulates extracellular polysaccharide production, biofilm formation, and rugose colony development by vibrio vulnificus.vibrio vulnificus is a human and animal pathogen that carries the highest death rate of any food-borne disease agent. it colonizes shellfish and forms biofilms on the surfaces of plankton, algae, fish, and eels. greater understanding of biofilm formation by the organism could provide insight into approaches to decrease its load in filter feeders and on biotic surfaces and control the occurrence of invasive disease. the capsular polysaccharide (cps), although essential for virulence, is not requi ...200818487410
[septic shock after exposure to vibrio vulnificus].vibrio vulnificus (vv) is a rare cause of infections in denmark. in this case, a patient suffering from chronic diseases developed wound infection, severe sepsis and despite adequate treatment, septic shock after exposure to seawater and fish. the fact that the condition was caused by vv was not identified until the results of the blood cultures. vv is the cause of primary sepsis, wound infections and gastroenteritis. the infection is treated with fluid resuscitation, broad-spectrum antibiotics ...200818492457
necrotising fasciitis caused by vibrio vulnificus in a man with cirrhosis. 200818501855
on the origin and function of an insertion element vpai-1 specific to post-1995 pandemic vibrio parahaemolyticus strains.since 1995, new virulent strains of vibrio parahaemolyticus have emerged and spread throughout the world. these "pandemic" strains have four strain specific genomic islands (gis), which are considered to be potential factors of the pandemicity. we investigated the origin and function of 24 genes in the so-called vpai-1, one of the four gis, by searching homologs in various species in bacteria and archaea. of these 24 genes, two are found only in vibrio vulnificus cmcp6 and shewanella sp. mr-7. t ...200818506094
the ecology of vibrio vulnificus, vibrio cholerae, and vibrio parahaemolyticus in north carolina estuaries.while numerous studies have characterized the distribution and/or ecology of various pathogenic vibrio spp., here we have simultaneously examined several estuarine sites for vibrio vulnificus, v. cholerae, and v. parahaemolyticus. for a one year period, waters and sediment were monitored for the presence of these three pathogens at six different sites on the east coast of north carolina in the united states. all three pathogens, identified using colony hybridization and pcr methods, occurred in ...200818545963
gene expression of cold shock and other stress-related genes in vibrio vulnificus grown in pure culture under shellstock temperature control conditions.shellstock refrigeration after harvesting is recommended to prevent further increases in vibrio vulnificus numbers in oysters, but it could potentially induce a cold shock response in this bacterium. v. vulnificus was incubated at 35, 25, 20, and 15 degrees c and then subjected to 7.2 and 4 degrees c for 1 week. a cold-adaptation response that enhanced cell culturability was observed when cells were incubated at 15 degrees c prior to cold shock at 7.2 degrees c. in vitro cold shock gene expressi ...200818236677
molecular cloning and sequence analysis of a novel zinc-metalloprotease gene from the salinivibrio sp. strain af-2004 and its extracellular expression in e. coli.in this work the first protease gene encoding a novel zinc-metalloprotease from the moderately halophilic bacterium salinivibrio sp. strain af-2004 has been cloned, sequenced and reported to the genbank. we have generated a library containing about 10,000 transformants whose screening yielded one clone harboring plasmid pbluescript with 3.6 kb inserted fragment (pbluesvp2) with positive caseinolytic activity. nucleotide sequence analysis of the selected clone revealed a single open reading frame ...200818093752
vibrio vulnificus rtx toxin kills host cells only after contact of the bacteria with host cells.vibrio vulnificus causes acute cell death and a fatal septicaemia. in this study, we show that contact with host cells is a prerequisite to the acute cytotoxicity. we screened transposon mutants defective in the contact-dependent cytotoxicity. two mutants had insertions within two open reading frames in a putative rtx toxin operon, the rtxa1 or rtxd encoding an rtx toxin (4701 amino acids) or an abc type transporter (467 amino acids). an rtxa1 mutation resulted in a cytotoxicity defect, which wa ...200818005241
role of the virulence plasmid pr99 and the metalloprotease vvp in resistance of vibrio vulnificus serovar e to eel innate immunity.vibrio vulnificus biotype 2 serovar e (vse) is a bacterial pathogen that produces a haemorrhagic septicaemia called vibriosis in eels. its ability to grow in blood is conferred by a recently described virulence plasmid [lee ct, amaro c, wu km, valiente e, chang yf, tsai sf, et al. a common virulence plasmid in biotype 2 vibrio vulnificus and its dissemination aided by a conjugal plasmid. journal of bacteriology, submitted for publication.]. in this study, we analyzed the role of this plasmid tog ...200818053741
survival of and in situ gene expression by vibrio vulnificus at varying salinities in estuarine environments.the opportunistic human pathogen vibrio vulnificus survives in a wide range of ecological environments, which demonstrates its ability to adapt to highly variable conditions. survival and gene expression under various conditions have been extensively studied in vitro; however, little work has been done to evaluate this bacterium in its natural habitat. therefore, this study monitored the long-term survival of v. vulnificus in situ and simultaneously evaluated the expression of stress (rpos, rela ...200817993554
evolution of exceptionally large genes in prokaryotes.analysis of bacterial genomic sequences revealed an average bacterial gene size of approximately 1 kb. however, genes with a size >10 kb were also noted. this study investigates the prevalence, possible function, and origin of exceptionally large-size genes (elsgs; >10 kb) in prokaryotes. forty-two elsgs (0.03%) were identified after searching more than 170,000 genes in 46 bacterial and 11 archaeal species. these elsgs were found in diverse species including both archaeal and eubacterial kingdom ...200818322635
characterization of the low-salinity stress in vibrio vulnificus.vibrio vulnificus is a marine pathogenic bacterium commonly found in seawater or seafood. this organism encounters low-salinity stress in its natural environment and during food processing. this study was designed to investigate the response of v. vulnificus yj03 to lethal low salinity (0.04% nacl) and its adaptation to sublethal salinity (0.12% nacl with 20 amino acids added). a short period in the nonculturable state was induced by lethal low-salinity stress followed by cell death after 30 min ...200818326198
evidence that aphb, essential for the virulence of vibrio vulnificus, is a global regulator.the vibrio vulnificus aphb mutant was significantly less virulent than the wild type and was impaired in motility and adherence to host cells. microarray analysis revealed that aphb of v. vulnificus (aphb(vv)) influences the expression of over 10% of the v. vulnificus genome. the combined results indicated that aphb(vv) is a global regulator contributing to the pathogenesis of v. vulnificus.200818344367
vibrio vulnificus rtxe is important for virulence, and its expression is induced by exposure to host cells.numerous secreted virulence factors have been proposed to account for the fulminating and destructive nature of vibrio vulnificus infections. a mutant of v. vulnificus that exhibited less cytotoxicity to int-407 human intestinal epithelial cells was screened from a library of mutants constructed by random transposon mutagenesis. a transposon-tagging method was used to identify and clone an open reading frame encoding an rtx toxin secretion atp binding protein, rtxe, from v. vulnificus. the deduc ...200818250174
autopsy cases of fulminant-type bacterial infection with necrotizing fasciitis: group a (beta) hemolytic streptococcus pyogenes versus vibrio vulnificus infection.two autopsy cases of fulminant-type infection associated with necrotizing fasciitis were analyzed clinicopathologically. both cases involved 57-year-old alcohol abusers. the former was a woman with group a (beta) hemolytic streptococcus pyogenes infection, and the latter was a man with vibrio vulnificus infection. the sudden onset of shock with high fever resulted in sepsis, decreased clotting, and hepatorenal symptoms, followed by death within a few days. post-mortem examination showed widespre ...200818251785
the crucial amino acid residue related to inactivation of vibrio vulnificus hemolysin.vibrio vulnificus, an opportunistic human pathogen causing fetal septicemia, produces a 50-kda pore-forming toxin as a virulence factor. this toxin consists of 451 amino acid residues; however, there are two types of this toxin on the basis of the difference of some amino acid residues, type 1 (leu(281), ser(415), asn(435)/asp(435), asn(438)) and type 2 (ile(281), asn(415), asn(435), thr(438)). in the present study, two characteristic properties of type 2 toxin that was elaborated by v. vulnific ...200817897806
role of the metalloprotease vvp and the virulence plasmid pr99 of vibrio vulnificus serovar e in surface colonization and fish virulence.the virulence for eels of vibrio vulnificus biotype 2 serovar e (vse) is conferred by a plasmid that codifies ability to survive in eel serum and cause septicaemia. to find out whether the plasmid and the selected chromosomal gene vvp plays a role in the initial steps of infection, the vse strain cect4999, the cured strain ct218 and the vvp-deficient mutant ct201 (obtained in this work by allelic exchange) were used in colonization and virulence experiments. the eel avirulent biotype 1 (bt1) str ...200818028416
variation of extracellular proteases produced by vibrio vulnificus clinical isolates: genetic diversity of the metalloprotease gene (vvp), and serine protease secretion by vvp-negative strains.vibrio vulnificus is a causative agent of septicemia or wound infection in human and eel; however, the genetic variation between human and eel isolates has been reported. in the present study, the difference in the vvp gene encoding a tissue-damaging metalloprotease was investigated. the gene of strain e86 from a diseased eel (type b vvp) was 95.2% identical with that of strain l-180 from human blood (type a vvp). pcr using oligonucleotide primers designed to differentiate two types of the gene ...200818262748
a common virulence plasmid in biotype 2 vibrio vulnificus and its dissemination aided by a conjugal plasmid.strains of vibrio vulnificus, a marine bacterial species pathogenic for humans and eels, are divided into three biotypes, and those virulent for eels are classified as biotype 2. all biotype 2 strains possess one or more plasmids, which have been shown to harbor the biotype 2-specific dna sequences. in this study we determined the dna sequences of three biotype 2 plasmids: pr99 (68.4 kbp) in strain cect4999 and pc4602-1 (56.6 kb) and pc4602-2 (66.9 kb) in strain cect4602. plasmid pc4602-2 showed ...200818156267
the crystal structure of a binary complex of two pseudopilins: epsi and epsj from the type 2 secretion system of vibrio vulnificus.type ii secretion systems (t2ss) translocate virulence factors from the periplasmic space of many pathogenic bacteria into the extracellular environment. the t2ss of vibrio cholerae and related species is called the extracellular protein secretion (eps) system that consists of a core of multiple copies of 11 different proteins. the pseudopilins, epsg, epsh, epsi, epsj and epsk, are five t2ss proteins that are thought to assemble into a pseudopilus, which is assumed to interact with the outer mem ...200818022192
epidemiologic study of vibrio vulnificus infections by using variable number tandem repeats.a 3-year environmental and clinical vibrio vulnificus survey using simple-sequence repeats typing shows that v. vulnificus biotype 3 constitutes approximately 21% of the bacterium population in tested aquaculture ponds as opposed to approximately 86% of clinical cases. simple-sequence repeats proved to be a useful epidemiologic tool, providing information on the environmental source of the pathogen.200919751592
dynamic expressions of liver tissue apoptosis-related genes of vibrio vulnificus sepsis rats and the effects of antibacterial agents.dynamic changes in mrna expressions of liver tissue apoptosis-promoting genes fas and bax and apoptosis-inhibiting gene bcl-2 of vibrio vulnificus sepsis rats were detected and the effects of antibacterial agents were examined. the rat model with vibrio vulnificus sepsis (vv group) was established and some of the vibrio vulnificus sepsis rats were treated with antibacterial agents (aa group). the mrna expressions of fas, bax and bcl-2 were measured by reverse transcription polymerase chain react ...200919399403
effect of antimicrobial agents on the toll-like receptors and inflammatory cytokines in liver tissue of the alcohol-induced liver disease in rats with vibrio vulnificus sepsis.septicemia and inflammation-mediated septic shock caused by vibrio vulnificus (vv) is strongly associated with chronic liver disease. this study examined the effects of antimicrobial therapy on expression of hepatic toll-like receptors and inflammatory cytokines in rats with alcohol-induced liver disease complicated by vv sepsis.200919781370
role of calcium/calmodulin signaling pathway in vibrio vulnificus cytolysin-induced hyperpermeability.endothelial hyperpermeability, a hallmark of septicemia, is induced by stress fiber formation, which is primarily regulated by the calcium/calmodulin signaling pathway in endothelial cells. we previously reported that trifluoperazine, a calcium/calmodulin antagonist, blocks vibrio vulnificus cytolysin (vvc) -induced lethality at in vivo animal model. the object of this study was therefore to examine whether vvc induces stress fiber formation through calcium/calmodulin signaling in endothelial ce ...200919447166
immune response and inhibition of bacterial growth by electrotransfer of plasmid dna containing the antimicrobial peptide, epinecidin-1, into zebrafish muscle.bacterial infections represent serious diseases in aquaculture, rapidly leading to fish death by septicemia. we investigated whether the electrotransfer of green fluorescent protein gene fusion epinecidin-1 (cmv-gfp-epi) dna into zebrafish muscle could regulate the fish immune response and inhibit bacterial growth. electroporation parameters such as the number of pulses, voltage, and amount of plasmid dna were analyzed, and results demonstrated the greatest mrna expression level of gfp-epi relat ...200919340938
[necrotizing fasciitis caused by a vibrio vulnificus infection in an eel fishery]. 200919348100
benthic ecology of vibrio spp. and pathogenic vibrio species in a coastal mediterranean environment (la spezia gulf, italy).we carried out a 16-month in situ study to investigate the ecology of vibrio spp. and pathogenic vibrio species in coastal sediments of the mediterranean sea, employing multiple-regression analysis to reveal the major environmental factors controlling their occurrence in the benthic environment. in addition, association between vibrios and sediment-inhabiting meiofauna, which is a major component of benthic ecosystems, was investigated. culturable and total vibrio spp. estimates by most-probable ...200919543938
tropical dermatology: marine and aquatic dermatology.dermatoses caused by marine organisms are frequently seen in dermatology clinics worldwide. cutaneous injuries after exposure to marine environments include bacterial and fungal infections and lesions caused by aquatic plants and protists. some of these diseases are well known by dermatologists, such as vibrio vulnificus septicemia and erysipeloid, but others are uncommon, such as envenomation caused by ingestion or contact with certain dinoflagellates or cyanobacteria, which are associated with ...200919836641
comparative evolutionary analysis of the major structural subunit of vibrio vulnificus type iv pili.type iv pili contribute to virulence in vibrio vulnificus, the bacterium responsible for the majority of fatal seafood-related infections. here, we performed within- and between-species evolutionary analysis of the gene that encodes the major structural subunit of the pilus, pila, by comparing it with pild and gyrb, the genes encoding the type iv prepilin peptidase and beta subunit of dna gyrase, respectively. although the diversity in pild and gyrb is similar to each other and likely to have ac ...200919556347
the capability of catabolic utilization of n-acetylneuraminic acid, a sialic acid, is essential for vibrio vulnificus pathogenesis.n-acetylneuraminic acid (neu5ac, sialic acid) could provide a good substrate for enteropathogenic bacteria in the intestine, when the bacteria invade and colonize in human gut. in order to analyze the role of neu5ac catabolism in vibrio vulnificus pathogenesis, a mutant with disruption of the nana gene encoding neu5ac lyase was constructed by allelic exchanges. the nana mutant was not able to utilize neu5ac as a sole carbon source and revealed an altered colony morphotype with reduced opacity in ...200919487477
hemochromatosis and vibrio vulnificus wound infections.there are several reports of persons with hemochromatosis and vibrio vulnificus primary septicemia, but few accounts of persons with hemochromatosis and v. vulnificus wound infection. a 58-year-old white man developed infection of a forearm injury exposed to seawater in the gulf of mexico near the alabama coast. at age 66, he was diagnosed to have hemochromatosis. transferrin saturation was 89% and serum ferritin was 4761 pmol/l. hfe genotype was c282y/c282y. serum levels of hepatic enzymes, glu ...200919349902
an amphioxus tlr with dynamic embryonic expression pattern responses to pathogens and activates nf-kappab pathway via myd88.a big bang expansion of the vertebrate-type (v-type) tlrs was reported in amphioxus. to shed lights on its implications, a unique tlr which is reversely inserted into an intron of amphioxus psmb7-10 by retrotransposition in the highly polymorphic proto-mhc region was cloned from chinese amphioxus (branchiostoma belcheri tsingtauense) and named as bbttlr1. in situ assays showed that bbttlr1 was predominantly expressed in pharynx and gut from larva to adult stages, which are considered as the firs ...200919394698
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