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immunogenicity and efficacy of highly purified invasin complex vaccine from shigella flexneri 2a.development of a subunit vaccine for shigellosis requires identification of protective antigens and delivering these antigens in a manner that stimulates immunity comparable to that induced by natural infection. the shigella invasin complex (invaplex) vaccine is an ion-exchange-purified extract from virulent shigella that consists of lps and several other proteins, including the invasins ipab and ipac. intranasal delivery of invaplex stimulates protective immunity in small animal models for shig ...200818276045
distribution and antibiotic susceptibility of shigella isolates in bangui, central african republic.we conducted a prospective study in four urban health care centres between january 2004 and november 2005 to determine the distribution and susceptibility patterns of shigella strains causing invasive diarrhoea in bangui, central african republic. of the 155 shigella isolated, shigella flexneri (51%) and shigella dysenteriae (30%) were the most common and the most resistant to usual antibiotics, including amoxicillin, sulphamethoxazole-trimethoprim and chloramphenicol. though multi-drug resistan ...200818282240
the shf gene of a shigella flexneri homologue on the virulent plasmid paa2 of enteroaggregative escherichia coli 042 is required for firm biofilm formation.enteroaggregative escherichia coli (eaec) is an increasingly important cause of diarrhea in both developing and industrialized countries, and is characterized by strong biofilm formation on the intestinal mucosa. sequencing of the virulent plasmid paa2 of the prototype eaec 042 revealed a cluster of three open reading frames (orfs; shf, capu, and virk) ca. 93% identical to a similar cluster located in shigella flexneri. the function of the first orf shf protein is not known, but the closest well ...200818293034
terpenoid compositions, and antioxidant and antimicrobial properties of the rhizome essential oils of different hedychium species.a phytochemical study of the rhizome essential oils of four different hedychium species was performed by means of gc and gc/ms analyses. h. ellipticum mainly contained 1,8-cineole, sabinene, and terpin-4-ol, while h. aurantiacum possessed terpin-4-ol, para-cymene, and bornyl acetate as the major entities. similarly, trans-meta-mentha-2,8-diene and linalool were noticed in h. coronarium. three different collections (i-iii) of h. spicatum showed amazing differences in the relative contents of thei ...200818293443
single residue determines the specificity of neutrophil elastase for shigella virulence factors.human neutrophil elastase (ne) is a key host defense protease that cleaves virulence factors of gram-negative bacteria. ne and cathepsin g (cg) are chymotrypsin-like serine proteases with sequence and structural similarities, and both are abundant in neutrophil granules. unlike ne, cg does not cleave virulence factors of enteric bacteria. through structure-function analysis, we identified regions in ne that are essential for cleaving shigella virulence proteins. ne residues at eight different po ...200818295791
geographic analysis of shigellosis in vietnam.geographic and ecological analysis may provide investigators useful ecological information for the control of shigellosis. this paper provides distribution of individual shigella species in space, and ecological covariates for shigellosis in nha trang, vietnam. data on shigellosis in neighborhoods were used to identify ecological covariates. a bayesian hierarchical model was used to obtain joint posterior distribution of model parameters and to construct smoothed risk maps for shigellosis. neigh ...200818296100
structures of the shigella flexneri type 3 secretion system protein mxic reveal conformational variability amongst homologues.many gram-negative pathogenic bacteria use a complex macromolecular machine, known as the type 3 secretion system (t3ss), to transfer virulence proteins into host cells. the t3ss is composed of a cytoplasmic bulb, a basal body spanning the inner and outer bacterial membranes, and an extracellular needle. secretion is regulated by both cytoplasmic and inner membrane proteins that must respond to specific signals in order to ensure that virulence proteins are not secreted before contact with a euk ...200818304577
ipgb1 and ipgb2, two homologous effectors secreted via the mxi-spa type iii secretion apparatus, cooperate to mediate polarized cell invasion and inflammatory potential of shigella flexenri.type iii secretion systems (t3ss) are present in many pathogenic gram-negative bacteria and mediate the translocation of bacterial effector proteins into host cells. here, we report the phenotypic characterization of s. flexneri ipgb1 and ipgb2 mutants, in which the genes encoding the ipgb1 and ipgb2 effectors have been inactivated, either independently or simultaneously. like ipgb1, we found that ipgb2 is secreted by the t3ss and its secretion requires the spa15 chaperone. upon infection of sem ...200818316224
delivery of biologically active anti-inflammatory cytokines il-10 and il-1ra in vivo by the shigella type iii secretion apparatus.pathogenicity of many gram-negative bacteria relies on a type iii secretion (t3s) apparatus, which is used for delivery of bacterial effectors into the host cell cytoplasm allowing the bacteria to manipulate host cell cytoskeleton network as well as to interfere with intracellular signaling pathways. in this study, we investigated the potential of the shigella flexneri t3sa as an in vivo delivery system for biologically active molecules such as cytokines. the anti-inflammatory cytokines il-10 an ...200818322242
impact of infections and normal flora in nonhuman primates on drug development.preclinical safety studies that are required for the marketing approval of a pharmaceutical include single and repeat dose studies in rodent and nonrodent species. the use of nonhuman primates (nhps), primarily macaques, as the nonrodent species has increased in recent years, in part due to the increase in development of biopharmaceuticals and immunomodulatory agents. depending on the source of the macaques, they may vary in genetic background, normal flora, and/or the incidence of preexisting p ...200818323580
cold shock exoribonuclease r (vacb) is involved in aeromonas hydrophila pathogenesis.in this study, we cloned and sequenced a virulence-associated gene (vacb) from a clinical isolate ssu of aeromonas hydrophila. we identified this gene based on our recently annotated genome sequence of the environmental isolate atcc 7966(t) of a. hydrophila and the vacb gene of shigella flexneri. the a. hydrophila vacb protein contained 798 amino acid residues, had a molecular mass of 90.5 kda, and exhibited an exoribonuclease (rnase r) activity. the rnase r of a. hydrophila was a cold-shock pro ...200818344363
maturation of paneth cells induces the refractory state of newborn mice to shigella infection.the intestinal tract of adult mice is naturally resistant to infection by shigella, the causative agent of bacillary dysentery in humans. conversely, newborn mice are highly susceptible to intragastric shigella infection and develop inflammatory lesions of the jejunal mucosa, very similar to those observed in the colon of dysenteric patients. however, the susceptibility period is short and one week after birth, animals have acquired a status of resistance characteristic of adult animals. to iden ...200818354217
differential expression of ompc and ompf in multidrug-resistant shigella dysenteriae and shigella flexneri by aqueous extract of aegle marmelos, altering its susceptibility toward beta-lactam antibiotics.steadily increasing resistance among shigella to beta-lactams, aminoglycosides, and tetracycline has compromised the utility of these commonly used antimicrobial agents. also, undesirable side effects of certain antibiotics have triggered immense interest in search of alternative therapies using medicinal plants. one such medicinal plant used since ancient times to cure diarrhea is aegle marmelos. the present study exemplifies the susceptibility of beta-lactam-resistant shigella dysenteriae and ...200818358664
differential regulation by magnesium of the two msbb paralogs of shigella flexneri.shigella flexneri, a gram-negative enteric pathogen, is unusual in that it contains two nonredundant paralogous genes that encode the myristoyl transferase msbb (lpxm) that catalyzes the final step in the synthesis of the lipid a moiety of lipopolysaccharide. msbb1 is encoded on the chromosome, and msbb2 is encoded on the large virulence plasmid present in all pathogenic shigellae. we demonstrate that myristoyl transferase activity due to msbb2 is detected in limited magnesium medium, but not in ...200818359815
analysis of rho-gtpase mimicry by a family of bacterial type iii effector proteins.microbial pathogens may hijack the actin cytoskeleton machinery to facilitate actin-based motility, cellular invasion, and intracellular trafficking through the endocytic pathway. of particular interest has been a large class of virulence factors collectively termed the type iii "effector" proteins. following contact with a eukaryotic cell, gram-negative bacterial pathogens employ a dedicated protein secretion apparatus termed type iii to translocate effector proteins into the cellular cytoplasm ...200818374161
coiled-coil regions play a role in the function of the shigella flexneri o-antigen chain length regulator wzzphs2.regulation of the length of the o-antigen (oag) chain attached to lps in shigella flexneri is important for virulence and is dependent on the inner-membrane protein wzz. a lack of high-resolution structural data for wzz has hampered efforts so far to correlate mutations affecting function of wzz with structure and describe a mechanism for chain length regulation. here we have used secondary structure prediction to show that the periplasmic domain of the wzz(phs2) protein has three regions of sig ...200818375803
ospf directly attenuates the activity of extracellular signal-regulated kinase during invasion by shigella flexneri in human dendritic cells.shigella spp., gram-negative pathogenic bacteria, deliver various effector molecules into the host cell cytoplasm through their type iii secretion system to facilitate their invasive process and control the host innate immune responses. although the function of these effectors is well characterized in epithelial cells during shigella infection, it has not been elucidated in the dendritic cell (dc), a major antigen presenting cell playing an important role in the initiation of immune responses. i ...200818378312
a pharmacokinetic study of enrofloxacin and its active metabolite ciprofloxacin after oral and intramuscular dosing of enrofloxacin in rhesus monkeys (macaca mulatta).enrofloxacin is used for treating shigellosis in non-human primates; however, there are no reports describing its pharmacokinetics in rhesus monkeys. pharmacokinetic data in intended target species (rhesus) help to determine the proper dose regimen. blood levels of enrofloxacin and ciprofloxacin (enrofloxacin's active metabolite), were determined after either intramuscular or oral dosing of enrofloxacin for 7 days in a cross-over study. levels of both antibiotics were determined by solid phase e ...200818384536
analysis of bacterial dna in synovial tissue of tunisian patients with reactive and undifferentiated arthritis by broad-range pcr, cloning and sequencing.bacteria and/or their antigens have been implicated in the pathogenesis of reactive arthritis (rea). several studies have reported the presence of bacterial antigens and nucleic acids of bacteria other than those specified by diagnostic criteria for rea in joint specimens from patients with rea and various arthritides. the present study was conducted to detect any bacterial dna and identify bacterial species that are present in the synovial tissue of tunisian patients with reactive arthritis and ...200818412942
containment of tumor-colonizing bacteria by host neutrophils.administration of facultative anaerobic bacteria like salmonella typhimurium, shigella flexneri, and escherichia coli to tumor-bearing mice leads to a preferential accumulation and proliferation of the microorganisms within the solid tumor. until now, all known tumor-targeting bacteria have shown poor dissemination inside the tumors. they accumulate almost exclusively in large necrotic areas and spare a rim of viable tumor cells. interestingly, the bacteria-containing necrotic region is separate ...200818413765
structure and genetics of shigella o antigens.this review covers the o antigens of the 46 serotypes of shigella, but those of most shigella flexneri are variants of one basic structure, leaving 34 shigella distinct o antigens to review, together with their gene clusters. several of the structures and gene clusters are reported for the first time and this is the first such group for which structures and dna sequences have been determined for all o antigens. shigella strains are in effect escherichia coli with a specific mode of pathogenicity ...200818422615
role of rna polymerase sigma-factor (rpos) in induction of glutamate-dependent acid-resistance of escherichia albertii under anaerobic conditions.escherichia albertii is a potential enteric food-borne pathogen with poorly defined genetic and biochemical properties. acid resistance is perceived to be an important property of enteric pathogens, enabling them to survive passage through stomach acidity so that they may colonize the mammalian gastrointestinal tract. we analyzed glutamate-dependent acid-resistance pathway (gdar) in five e. albertii strains that have been identified so far. we observed that the strains were unable to induce gdar ...200818422618
comparing the antimicrobial effectiveness of nacl and kcl with a view to salt/sodium replacement.a study using a small range of pathogenic bacterial species (aeromonas hydrophila, enterobacter sakazakii, shigella flexneri, yersinia enterocolitica and 3 strains of staphylococcus aureus) has shown that potassium chloride has an equivalent antimicrobial effect on these organisms when calculated on a molar basis. combined nacl and kcl experiments were carried out and data was analysed using a modification to the lambert and lambert [lambert, r.j.w., and lambert, r., 2003. a model for the effica ...200818423764
virulent shigella flexneri subverts the host innate immune response through manipulation of antimicrobial peptide gene expression.antimicrobial factors are efficient defense components of the innate immunity, playing a crucial role in the intestinal homeostasis and protection against pathogens. in this study, we report that upon infection of polarized human intestinal cells in vitro, virulent shigella flexneri suppress transcription of several genes encoding antimicrobial cationic peptides, particularly the human beta-defensin hbd-3, which we show to be especially active against s. flexneri. this is an example of targeted ...200818426984
alternative hand contamination technique to compare the activities of antimicrobial and nonantimicrobial soaps under different test conditions.antimicrobial hand soaps provide a greater bacterial reduction than nonantimicrobial soaps. however, the link between greater bacterial reduction and a reduction of disease has not been definitively demonstrated. confounding factors, such as compliance, soap volume, and wash time, may all influence the outcomes of studies. the aim of this work was to examine the effects of wash time and soap volume on the relative activities and the subsequent transfer of bacteria to inanimate objects for antimi ...200818441107
shigellosis: the current status of vaccine development.shigellosis, a major form of bacillary dysentery, is caused by infection with shigella organisms. in poor countries, shigella-caused dysentery is endemic and causes an estimated 163 million illness episodes annually and more than one million deaths. although several strategies have been used to develop vaccines targeting shigellosis, none has been licensed for use outside china. owing to the wide range of shigella serotypes and subtypes, there is a need for a multivalent vaccine representing pre ...200818448978
deoxycholate interacts with ipad of shigella flexneri in inducing the recruitment of ipab to the type iii secretion apparatus needle tip.type iii secretion (tts) is an essential virulence function for shigella flexneri that delivers effector proteins that are responsible for bacterial invasion of intestinal epithelial cells. the shigella tts apparatus (ttsa) consists of a basal body that spans the bacterial inner and outer membranes and a needle exposed at the pathogen surface. at the distal end of the needle is a "tip complex" composed of invasion plasmid antigen d (ipad). ipad not only regulates tts, but is required for the rec ...200818450744
mutagenesis of the shigella flexneri autotransporter icsa reveals novel functional regions involved in icsa biogenesis and recruitment of host neural wiscott-aldrich syndrome protein.the icsa (virg) protein of shigella flexneri is a polarly localized, outer membrane protein that is essential for virulence. within host cells, icsa activates the host actin regulatory protein, neural wiskott-aldrich syndrome protein (n-wasp), which in turn recruits the arp2/3 complex, which nucleates host actin to form f-actin comet tails and initiate bacterial motility. linker insertion mutagenesis was undertaken to randomly introduce 5-amino-acid in-frame insertions within icsa. forty-seven l ...200818456802
an intersubunit active site between supercoiled parallel beta helices in the trimeric tailspike endorhamnosidase of shigella flexneri phage sf6.sf6 belongs to the podoviridae family of temperate bacteriophages that infect gram-negative bacteria by insertion of their double-stranded dna. they attach to their hosts specifically via their tailspike proteins. the 1.25 a crystal structure of shigella phage sf6 tailspike protein (sf6 tsp) reveals a conserved architecture with a central, right-handed beta helix. in the trimer of sf6 tsp, the parallel beta helices form a left-handed, coiled-beta coil with a pitch of 340 a. the c-terminal domain ...200818462681
immunological evidence for functional rather than structural mimicry by a shigella flexneri y polysaccharide-mimetic peptide.an approach to vaccine design is the use of molecules that mimic the immunogenic element of interest. in this context, the interaction of mdwnmhaa, a peptide mimic of the shigella flexneri y o polysaccharide (ps), with an anti-carbohydrate monoclonal antibody, as studied previously by x-ray crystallography, suggested the presence of functional rather than structural mimicry and a bound peptide conformation that was not represented significantly in the free-ligand ensemble. the antibody response ...200818463226
simple sequence repeats in different genome sequences of shigella and comparison with high gc and at-rich genomes.simple sequence repeats (ssrs) are omnipresent in prokaryotes and eukaryotes, and are found anywhere in the genome in both protein encoding and noncoding regions. in present study the whole genome sequences of seven chromosomes (shigella flexneri 2a str301 and 2457t, shigella sonnei, escherichia coli k12, mycobacterium tuberculosis, mycobacterium leprae and staphylococcus saprophyticus) have downloaded from the genbank database for identifying abundance, distribution and composition of ssrs and ...200818464038
synthesis, characterization and antibacterial activity of cobalt(iii) complexes with pyridine-amide ligands.the ligands 2-(n-(x-pyridyl)carbamoyl)pyridine (x=2, 3 or 4 for hl1-hl3, respectively) and 2,6-bis(n-(y-pyridyl)carbamoyl)pyridine (y=2, 3 or 4 for h2l4-h2l6, respectively) in their mono- and di-deprotonated forms have been used to synthesize kinetically stable cobalt(iii) compounds [co(l1-3)3] (1-3) and na[co(l4-6)2] (4-6), respectively. the co(iii) ion is in octahedral environment and is surrounded by three bidentate ligands in complexes 1-3 and two tridentate ligands in complexes 4-6. ligands ...200817959275
identification of newly recognized serotype 1c as the most prevalent shigella flexneri serotype in northern rural vietnam.we investigated the identity of 37 shigella flexneri strains that had previously been isolated from northern rural vietnam (son tay province) and described as untypable. twenty-four isolates reacted with masf 1c, a monoclonal antibody specific for s. flexneri serotype 1c. a further ten untypable isolates were found to be rough mutants (no longer expressing o-antigen) that were derived from serotype 1c strains. pulsed-field gel electrophoresis demonstrated that these strains consisted of many dif ...200817922932
a role for nod-like receptors in autophagy induced by shigella infection.shigella infection, the cause of bacillary dysentery, induces caspase-1 activation and cell death in macrophages, but the precise mechanisms remain poorly understood. in our recent study, we presented evidence that caspase-1 activation and il-1beta processing induced by shigella are mediated through ipaf, a cytosolic pattern-recognition receptor of the nucleotide-binding oligomerization domain (nod)-like receptor (nlr) family and the adaptor protein apoptosis-associated speck-like protein contai ...200817932464
isolation and antibiotic susceptibility of salmonella and shigella strains isolated from children in gaza, palestine from 1999 to 2006.diarrhea and acute gastroenteritis are among the leading causes of illnesses and death in infants and children throughout the world, especially in developing countries, globally, salmonella and shigella remain the major contributors to acute enteric infections and diarrhea.200817854424
risk areas and neighborhood-level risk factors for shigella dysenteriae 1 and shigella flexneri.annually, there are 165 million cases of shigellosis resulting in 1.1 million deaths in the developing world. two species of the causative agent, shigella dysenteriae 1 and shigella flexneri, are responsible for acute diarrheal illness. vaccines are being developed against both but many questions remain about the disease burden, distribution of species in space and time, and community-level risk factors. this study answers two questions. what are the differences in neighborhood-level risk factor ...200817602851
systemic pharmacokinetics of rifaximin in volunteers with shigellosis.rifaximin is an oral antibiotic indicated for treatment of traveler's diarrhea. rifaximin pharmacokinetics were evaluated in individuals challenged with shigella flexneri. peak plasma rifaximin concentrations were low after nine consecutive doses, and no accumulation was observed. rifaximin serum levels were minimal and similar to those previously reported in studies of healthy volunteers.200818086855
saccharomyces boulardii interferes with shigella pathogenesis by postinvasion signaling events.saccharomyces boulardii is gaining in popularity as a treatment for a variety of diarrheal diseases as well as inflammatory bowel disease. this study was designed to examine the effect of this yeast on infection by shigella flexneri, a highly infectious and human host-adapted enteric pathogen. we investigated key interactions between the bacteria and host cells in the presence of the yeast in addition to a number of host responses including proinflammatory events and markers. although the presen ...200818032477
the pattern-recognition molecule nod1 is localized at the plasma membrane at sites of bacterial interaction.the pattern-recognition molecule nod1 is a critical sensor for bacterial derived diaminopimelic acid-containing peptidoglycan fragments which induces innate immune responses in epithelial cells. here we report the subcellular localization of this protein in human epithelial cells. nod1 is localized in the cytosol and at the plasma membrane in human cells. this membrane association is dependent on the integrity of the protein, on its signalling capacity and on an intact actin cytoskeleton. signal ...200817970764
the nlee/ospz family of effector proteins is required for polymorphonuclear transepithelial migration, a characteristic shared by enteropathogenic escherichia coli and shigella flexneri infections.enteropathogenic escherichia coli (epec) and shigella flexneri are human host-specific pathogens that infect intestinal epithelial cells. however, each bacterial species employs a different infection strategy within this environmental niche. epec attaches to the apical surface of small intestine enterocytes, causing microvillus effacement and rearrangement of the host cell cytoskeleton beneath adherent bacteria. in contrast, s. flexneri invades the large intestine epithelium at the basolateral m ...200817984206
intracellular bacteriolysis triggers a massive apoptotic cell death in shigella-infected epithelial cells.infected epithelial cells, which act as a first barrier against pathogens, seldom undergo apoptosis. rather, infected epithelial cells undergo a slow cell death that displays hallmarks of necrosis. here, we demonstrate that rapid intracellular lysis of shigella flexneri, provoked by either the use of a diaminopimelic acid auxotroph mutant or treatment of infected cells with antibiotics of the beta-lactam family, resulted in a massive and rapid induction of apoptotic cell death. this intracellula ...200818606244
selective activation of human intestinal mast cells by escherichia coli hemolysin.mast cells (mcs) are recognized to play an important role in bacterial host defense in the murine system. in this study, we studied the interaction of human mcs, isolated from the intestine and purified to homogeneity, with different escherichia coli and shigella flexneri strains. we show that alpha-hemolysin (hly)-producing e. coli strains induce the release of histamine, leukotrienes, and proinflammatory cytokines in intestinal mcs. in contrast, mcs were virtually unresponsive to s. flexneri a ...200818606698
safety and immunogenicity of wrsd1, a live attenuated shigella dysenteriae type 1 vaccine candidate.among shigella serotypes shigella dysenteriae type 1 produces the most severe disease, including cases of hemolytic-uremic syndrome and pandemic outbreaks. wrsd1 is a live s. dysenteriae 1 strain attenuated by deletion of the virg(icsa) gene, which encodes a protein that mediates intercellular spread, and stxa and stxb, which encode the shiga toxin. in this phase i trial five groups of eight subjects ingested escalating doses of wrsd1 ranging from 10(3) to 10(7)cfu. no subject experienced fever ...200818468742
structural and functional studies indicate that shigella vira is not a protease and does not directly destabilize microtubules.vira, an essential virulence factor in shigella disease pathogenesis, is involved in the uptake, motility, and cell-to-cell spread of shigella organisms within the human host. these functions have been attributed to a vira protease activity and a mechanism of microtubule destruction via tubulin degradation [yoshida, s., et al. (2006) science 314, 985-989]. we report functional and crystallographic data indicating a novel vira structure that lacks these activities but highlights the homology to t ...200818763811
bacterial conjugation in the cytoplasm of mouse cells.intracellular pathogenic organisms such as salmonellae and shigellae are able to evade the effects of many antibiotics because the drugs are not able to penetrate the plasma membrane. in addition, these bacteria may be able to transfer genes within cells while protected from the action of drugs. the primary mode by which virulence and antibiotic resistance genes are spread is bacterial conjugation. salmonellae have been shown to be competent for conjugation in the vacuoles of cultured mammalian ...200818765719
occurrence and characteristics of the cytolysin a gene in shigella strains and other members of the family enterobacteriaceae.cytolysin a (clya, hlye, shea) is a hemolytic pore-forming toxin found in escherichia coli and salmonella enterica serovars typhi and paratyphi a. in the present study, analysis of several shigella strains revealed that they harbor only nonfunctional clya gene copies that have been inactivated either by the integration of insertion sequence (is) elements (shigella dysenteriae, shigella boydii, and shigella sonnei strains) or by a frameshift mutation (shigella flexneri). shigella dysenteriae and ...200818754791
topology and identification of critical residues of the o-acetyltransferase of serotype-converting bacteriophage, sf6, of shigella flexneri.the modification of the lps o-antigen, seen in the diverse serotypes of shigella flexneri is brought about by the glucosyltransferases (gtr) and the o-acetyltransferase (oac). in this study, we establish the membrane topology of oac using the dual reporter phoa-laczalpha. we have determined that oac is an integral membrane protein with 10 transmembrane regions. the hydrophilic n- and c-termini are oriented in the cytoplasm. functionally important cytoplasmic and periplasmic loops have also been ...200818755141
[htpg protein of shigella flexneri 2a strain 2457t evokes inflammatory response in mice].to analyze the function of htpg of s. flexneri 2a 2457t, we constructed an htpg deletion mutant and a recovery mutant.200818837368
evaluation by real-time pcr of the expression of s. flexneri virulence-associated genes ospb and phon2 under different genetical backgrounds.under conditions of activated type iii secretion shigella flexneri up-regulates the expression of numerous genes, including the virulence plasmid (pinv)-encoded ospb and phon2 genes. ospb and phon2 are virulence-associated genes which are part of a bicistronic transcriptional unit encoding ospb, a protein (effector) of unknown function secreted by the type iii secretion (tts) apparatus, and phon2 (apyrase or atp-diphosphohydrolase), a periplasmic protein involved in polar icsa localization on th ...200818831939
novel fold of vira, a type iii secretion system effector protein from shigella flexneri.vira, a secreted effector protein from shigella sp., has been shown to be necessary for its virulence. it was also reported that vira might be related to papain-like cysteine proteases and cleave alpha-tubulin, thus facilitating intracellular spreading. we have now determined the crystal structure of vira at 3.0 a resolution. the shape of the molecule resembles the letter "v," with the residues in the n-terminal third of the 45-kda molecule (some of which are disordered) forming one clearly iden ...200818787201
incidence of bacterial enteropathogens among hospitalized diarrhea patients from orissa, india.bacteriological analysis of 1,551 stool/rectal swabs from all age groups of diarrhea patients of different hospitals of orissa from january 2004 to december 2006 was carried out using standard procedures. among all enteropathogens isolated in 886 culture-positive samples, escherichia coli constituted 75.5%, including 13.2% pathogenic e. coli; vibrio cholerae o1 constituted 17.3%; v. cholerae o139, 1%; shigella spp., 4.5% (shigella flexneri type 6, 2.9%, s. dysenteriae type i, 0.7%, s. sonnei, 0. ...200818806340
subproteomic tools to increase genome annotation complexity.comprehensive and precise annotations of short protein-coding genes are always a challenging task. here we propose a new design to facilitate the characterization of previously overlooked short protein-coding genes by integrating a shotgun proteomics method and oligonucleotide array analysis. using shigella flexneri as a model, we validate 163 annotated orfs and 51 hypothetical or putative transcripts at the protein level, and discover four novel short orfs. this strategy will contribute signifi ...200818814329
[analysis on incidence of bacillary dysentery in jinan municipality from 1951 to 2005].to explore the epidemic characteristics of bacterial dysentery in jinan municipality, and to provide scientific basis for effective strategy for bacterial dysentery control.200818844085
spa32 interaction with the inner-membrane spa40 component of the type iii secretion system of shigella flexneri is required for the control of the needle length by a molecular tape measure mechanism.the effectors of enterocyte invasion by shigella are dependent on a type iii secretion system that contains a needle whose length average does not exceed 50 nm. previously, we reported that spa32 is required for needle length control as well as to switch substrate specificity from mxih to ipa proteins secretion. to identify functional domains of spa32, 11 truncated variants were constructed and analysed for their capacity (i) to control the needle's length; (ii) to secrete the ipa proteins; and ...200819019157
structure of the shigella t3ss effector ipah defines a new class of e3 ubiquitin ligases.ipah proteins are e3 ubiquitin ligases delivered by the type iii secretion apparatus into host cells upon infection of humans by the gram-negative pathogen shigella flexneri. these proteins comprise a variable leucine-rich repeat-containing n-terminal domain and a conserved c-terminal domain harboring an invariant cysteine residue that is crucial for activity. ipah homologs are encoded by diverse animal and plant pathogens. here we demonstrate that the ipah c-terminal domain carries the catalyti ...200818997778
structure of a shigella effector reveals a new class of ubiquitin ligases.bacterial pathogens have evolved effector proteins with ubiquitin e3 ligase activities through structural mimicking. here we report the crystal structure of the shigella flexneri type iii effector ipah3, a member of the leucine-rich repeat (lrr)-containing bacterial e3 family. the lrr domain is structurally similar to yersinia pestis yopm and potentially binds to substrates. the structure of the c-terminal e3 domain differs from the typical ring- and hect-type e3s. ipah3 synthesizes a lys48-link ...200818997779
in this issue: proteomics 20/2008.in this issue of proteomics you will find the following highlighted articles:comprehensively confirmed annotation of short orfsduring the california gold rush, prospectors did not look only for large nuggets to pave their road to riches - they also collected gold dust. the proteomic equivalent of dust is the small orf. it does not pay to put a lower limit on dust or orf size because almost any size may be turned into "cash" or an important peptide. wei et al. demonstrate how a combination of con ...200818924177
a pilot sheds light on secretin assembly. 200818940600
structural characterization of the type-iii pilot-secretin complex from shigella flexneri.assembly of the type-iii secretion apparatus, which translocates proteins through both membranes of gram-negative bacterial pathogens into host cells, requires the formation of an integral outer-membrane secretin ring. typically, a small lipidated pilot protein is necessary for the stabilization and localization of this ring. using nmr spectroscopy, we demonstrate that the c-terminal residues 553-570 of the shigella flexneri secretin mxid encompass the minimal binding domain for its cognate pilo ...200818940609
detection of protozoan and bacterial pathogens of public health importance in faeces of corvus spp. (large-billed crow).parasites and bacteria are reported in the faeces of birds in the current study. fresh faecal samples of the large-billed crow (corvus spp.) were collected from the study site at bangsar, an urban setting in kuala lumpur, malaysia. these samples were transported to laboratory and analysed for parasites and bacteria. pre-prepared xld agar plates were used for culturing the bacteria in the laboratory. using the api 20etm test strips, 9 different species of bacteria were identified belonging to the ...200818948884
analysis of immune responses and serological cross reactivities among vibrio cholerae o1, shigella flexneri 2a and haemophilus influenzae b.antigenic determinants expressed on the bacterial cell surface are of importance in the serological characterization and microbiological diagnosis. the bacterial strains carrying these identical or similar antigenic epitopes might react with antibodies produced against other strains. in this study, strong immunogenicity and antigenic cross reactivity were demonstrated among v. cholerae o1, s. flexnerii 2a and h. influenzae b surface components. the enzyme linked immunosorbent assay (elisa) resul ...200818954564
full structural characterization of shigella flexneri m90t serotype 5 wild-type r-lps and its delta galu mutant: glycine residue location in the inner core of the lipopolysaccharide.shigella flexneri is a gram-negative bacterium responsible for serious enteric infections that occur mainly in the terminal ileum and colon. high interest in shigella, as a human pathogen, is driven by its antibiotic resistance and the necessity to develop a vaccine against its infections. vaccines of the last generation use carbohydrate moieties of the lipopolysaccharide as probable candidates. for this reason, the primary structure of the core oligosaccharide from the r-lps produced by s. flex ...200818174310
ph sensitivity of type iii secretion system tip proteins.many pathogenic gram-negative bacteria employ type iii secretion systems to transport proteins into the host cell membrane and cytoplasm to subvert normal cellular functions. the type iii secretion apparatus consists of a basal body spanning the inner and outer bacterial membranes and a needle which extends away from the bacterium. recent work has found that a special class of proteins localizes to the tip of the needle to control secretion of effector proteins. five of these tip proteins are ip ...200818175320
differential activation of human tlr4 by escherichia coli and shigella flexneri 2a lipopolysaccharide: combined effects of lipid a acylation state and tlr4 polymorphisms on signaling.the lipid a of lps activates tlr4 through an interaction with myeloid differentiation protein-2 (md-2) and the degree of lipid a acylation affects tlr4 responsiveness. two tlr4 single nucleotide polymorphisms (asp299gly and thr399ile) have been associated with lps hyporesponsiveness. we hypothesized that the combination of hypoacylation and these single nucleotide polymorphisms would exhibit a compounded effect on tlr4 signaling. hek293t transfectants expressing wild-type or polymorphic tlr4 wer ...200818178854
phenotypic and molecular characterization of tem-116 extended-spectrum beta-lactamase produced by a shigella flexneri clinical isolate from chickens.extended-spectrum beta-lactamases (esbls) produced by a clinical isolate of shigella flexneri from chickens were detected with confirmatory phenotypic tests of the clinical and laboratory standards institute, and minimum inhibitory concentrations of several antibacterial drugs against the isolate were determined by the twofold dilution method. the genotype and subtype of the esbl-producing s. flexneri isolate were identified by pcr amplifying of esbl genes and dna sequencing analysis. the result ...200818093137
muramylpeptide shedding modulates cell sensing of shigella flexneri.bacterial infections trigger the activation of innate immunity through the interaction of pathogen-associated molecular patterns (pamps) with pattern recognition molecules (prms). the nucleotide-binding oligomerization domain (nod) proteins are intracellular prms that recognize muramylpeptides contained in peptidoglycan (pgn) of bacteria. it is still unclear how nod1 physically interacts with pgn, a structure internal to the gram-negative bacterial envelope. to contribute to the understanding of ...200818042252
cytoplasmic targeting of ipac to the bacterial pole directs polar type iii secretion in shigella.type iii secretion (t3s) systems are largely used by pathogenic gram-negative bacteria to inject multiple effectors into eukaryotic cells. upon cell contact, these bacterial microinjection devices insert two t3s substrates into host cell membranes, forming a so-called 'translocon' that is required for targeting of type iii effectors in the cell cytosol. here, we show that secretion of the translocon component ipac of invasive shigella occurs at the level of one bacterial pole during cell invasio ...200818188151
bacterial actin assembly requires toca-1 to relieve n-wasp autoinhibition.actin polymerization in the mammalian cytosol can be locally activated by mechanisms that relieve the autoinhibited state of n-wasp, an initiator of actin assembly, a process that also requires the protein toca-1. several pathogenic bacteria, including shigella, exploit this host feature to infect and disseminate efficiently. the shigella outer membrane protein icsa recruits n-wasp, which upon activation at the bacterial surface mediates localized actin polymerization. the molecular role of toca ...200818191793
the chaperone ipgc copurifies with the virulence regulator mxie.the expression of a subset of shigella flexneri virulence genes is dependent upon a cytoplasmic chaperone, ipgc, and an activator from the arac/xyls family, mxie. in this paper, we report that the chaperone forms a specific and stable heteromer with mxie.200818192384
role and regulation of iron-sulfur cluster biosynthesis genes in shigella flexneri virulence.shigella flexneri, a causative agent of bacterial dysentery, possesses two predicted iron-sulfur cluster biosynthesis systems called suf and isc. s. flexneri strains containing deletion mutations in the entire suf operon (ur011) or the iscsua genes (ur022) were constructed. both mutants were defective in surviving exposure to oxidative stress. the suf mutant showed growth that was comparable to that of the parental strain in both iron-replete and iron-limiting media; however, the isc mutant show ...200818195027
molecular pathogenesis of shigella spp.: controlling host cell signaling, invasion, and death by type iii secretion.shigella spp. are gram-negative pathogenic bacteria that evolved from harmless enterobacterial relatives and may cause devastating diarrhea upon ingestion. research performed over the last 25 years revealed that a type iii secretion system (t3ss) encoded on a large plasmid is a key virulence factor of shigella flexneri. the t3ss determines the interactions of s. flexneri with intestinal cells by consecutively translocating two sets of effector proteins into the target cells. thus, s. flexneri co ...200818202440
phase 1 clinical trial of live attenuated shigella dysenteriae type-1 deltaicsa deltaent deltafep deltastxa:hgr oral vaccine sc599 in healthy human adult volunteers.twenty-eight adults received between 10(2) and 10(8)colony forming units of live shigella dysenteriae type-1 vaccine sc599, attenuated by deletion of invasion (icsa), iron chelation (ent, fep) and shiga toxin a-subunit (stxa) genes, followed by ciprofloxacin on day 4. dose-independent diarrhea or change in bowel habit was seen in 3 subjects, without dysentery, vaccinaemia or serious adverse events. hematology and biochemical parameters were unchanged. doses of 10(5) or greater induced dose-indep ...200818207287
antimicrobial susceptibility of shigella isolates in eight asian countries, 2001-2004.shigellosis is a major health problem in developing countries, causing 91 million episodes and 414,000 deaths in asia annually. because of increasing trends towards drug resistance, this study was undertaken to monitor local resistance patterns of shigella isolates from 8 asian countries.200818473096
crystal structure of spa40, the specificity switch for the shigella flexneri type iii secretion system.the pathogenic bacterium shigella flexneri uses a type iii secretion system to inject virulence factors from the bacterial cytosol directly into host cells. the machinery that identifies secretion substrates and controls the export of extracellular components and effector proteins consists of several inner-membrane and cytoplasmic proteins. one of the inner membrane components, spa40, belongs to a family of proteins proposed to regulate the switching of substrate specificity of the export appara ...200818485071
the response of type three secretion system needle proteins mxihdelta5, bsaldelta5, and prgidelta5 to temperature and ph.the type iii secretion system (ttss) is a specialized supramolecular injectisome composed of 25 or more proteins which form basal and extracellular domains and share gross architectural similarities with bacterial flagella. the extracellular component of the "needle complex" is primarily composed of a single monomeric subunit organized in a helical array surrounding a hollow pore and protrudes from the bacterial membrane. it is through this surface appendage that virulence factors are translocat ...200818491382
distinct isoforms of phospholipase a2 mediate the ability of salmonella enterica serotype typhimurium and shigella flexneri to induce the transepithelial migration of neutrophils.salmonella spp. and shigella spp. are responsible for millions of cases of enteric disease each year worldwide. while these pathogens have evolved distinct strategies for interacting with the human intestinal epithelium, they both induce significant proinflammatory responses that result in massive transepithelial migration of neutrophils across the intestinal mucosa. it has previously been shown with salmonella enterica serotype typhimurium that the process of neutrophil transmigration is mediat ...200818505810
the serine protease motif of pic mediates a dose-dependent mucolytic activity after binding to sugar constituents of the mucin substrate.the pic gene is harbored on the chromosomes of three important pathogens: enteroaggregative escherichia coli (eaec), uropathogenic e. coli (upec), and shigella flexneri. since pic is secreted into the intestinal lumen during eaec infection, we sought to identify intestinal-mucosal substrates for pic. pic did not damage epithelial cells, cleave fodrin, or degrade host defense proteins embedded in the mucus layer (siga, lactoferrin and lysozyme). however, by using a solid-phase assay to evaluate t ...200818538533
shigella bacteraemia over a decade in soweto, south africa.shigella infection rarely penetrates beyond the intestinal mucosa to cause bacteraemia. aids patients may be at increased risk. we investigated shigella bacteraemia rates over a decade in a south african population with a high hiv prevalence and analysed presentation and outcome. a retrospective cohort analysis was performed for hospitalized patients for 2003-2006. the rates were compared to those for 1996-1998. adult medical rates decreased from 0.195/1000 in 1996-1998 to 0.087/1000 admissions ...200818550134
molecular characterization of multidrug-resistant shigella species isolated from epidemic and endemic cases of shigellosis in india.shigella species represent one of the growing numbers of antimicrobial-resistant bacteria in developing countries. fluoroquinolone-resistant strains of shigella dysenteriae type 1 and shigella flexneri type 2a emerged in india during 2002 and 2003, respectively. sixty strains of shigella from different parts of india were analysed for antimicrobial susceptibility, the presence of the qnr plasmid, mutations in the quinolone resistance determining regions (qrdrs), fluoroquinolone accumulation, and ...200818566144
design, synthesis, characterization, antibacterial and antifungal activities of a novel class of 5,7-diaryl-4,4-dimethyl-4,5,6,7-tetrahydropyridino[3,4-d]-1,2,3-selenadiazoles.compound 26 is more potent against escherichia coli. and 24 is more active against staphylococcus aureus, beta-heamolytic streptococcus, vibreo cholerae, salmonella typhii, and shigella flexneri than the standard drug ciprofloxacin. moreover, of all the compounds tested, 26 is more effective against aspergillus flavus and mucor, than the standard drug fluconazole.200818569338
vibrio cholerae vcib promotes iron uptake via ferrous iron transporters.vibrio cholerae uses a variety of strategies for obtaining iron in its diverse environments. in this study we report the identification of a novel iron utilization protein in v. cholerae, vcib. the vcib gene and its linked gene, vcia, were isolated in a screen for v. cholerae genes that permitted growth of an escherichia coli siderophore mutant in low-iron medium. the vciab operon encodes a predicted tonb-dependent outer membrane receptor, vcia, and a putative inner membrane protein, vcib. vcib, ...200818586940
anti-infectious effect of s-benzylisothiourea compound a22, which inhibits the actin-like protein, mreb, in shigella flexneri.s-benzylisothiourea compound a22 induces coccoid forms in escherichia coli by inhibiting the function of the actin-like cytoskeletal protein, mreb. the minimum inhibitory concentration of a22 and the minimum concentration to induce coccoid forms for various pathogenic bacteria were determined. at 10 microg/ml, a22 induced coccoid forms in shigella flexneri but did not inhibit the growth. no alteration of coccoid forms in the gram-positive bacteria and anaerobic bacteria tested were observed foll ...200818591769
[epidemic strains of different plasmidovars of bacteria from shigella genus].plasmid analysis of 46 strains of shigella species confirmed the view about heterogeneity and variability of plasmids composition in these microorganisms. on the other hand,the presence of low-molecular, multiplicated extrachromosomal replicons, which were stable, was shown. using them, it was possible to divide the shigella isolates into two major plasmidovars, characterize them according to repertoire of the most stable plasmids and determine epidemically significant variants during sporadic a ...200818595459
[electrical resistance of bacterial culture suspension as indicator of epidemic and sporadic variants of shigella].for the first time method of measurement of electric resistance of bacterial suspension was used for differentiation of "epidemic" and "sporadic" (further used without quotes) variants of shigella species. study of 40 strains of s. flexneri and s. sonnei revealed significant differences in electric resistance of cultures isolated during epidemic outbreaks after the growth during 1 or 6 hours - 18.1+/-8.3 kohm and 84.6+/-7.8 kohm respectively (increase in 4.9-fold; p<0.01), while non significant ...200818597992
structures of synthetic o-antigen fragments from serotype 2a shigella flexneri in complex with a protective monoclonal antibody.the anti-lps igg mab f22-4, raised against shigella flexneri serotype 2a bacteria, protects against homologous, but not heterologous, challenge in an experimental animal model. we report the crystal structures of complexes formed between fab f22-4 and two synthetic oligosaccharides, a decasaccharide and a pentadecasaccharide that were previously shown to be both immunogenic and antigenic mimics of the s. flexneri serotype 2a o-antigen. f22-4 binds to an epitope contained within two consecutive 2 ...200818621718
crystal structure of a glutamate/aspartate binding protein complexed with a glutamate molecule: structural basis of ligand specificity at atomic resolution.the crystal structure of a periplasmic l-aspartate/l-glutamate binding protein (debp) from shigella flexneri complexed with an l-glutamate molecule has been determined and refined to an atomic resolution of 1.0 a. there are two debp molecules in the asymmetric unit. the refined model contains 4462 non-hydrogen protein atoms, 730 water molecules, 2 bound glutamate molecules, and 2 tris molecules from the buffer used in crystallization. the final r(cryst) and r(free) factors are 13.61% and 16.89%, ...200818640128
acute diarrhea due to enteropathogenic bacteria in patients at hospitals in tehran.during a study examining causes of diarrhea from may 2004 to may 2005, 808 stool specimens were collected from patients with acute diarrhea in tehran. fecal samples were cultured and identified according to the standard biochemical methods. molecular identification of enteropathogens was carried out by amplification of their virulence genes by polymerase chain reaction. a total of 369 (45.6%) bacterial pathogens were recovered from 808 patients as follows: shigella spp., 155 (45.6%); diarrheagen ...200818653967
antimicrobial resistance in shigella species isolated in dakar, senegal (2004-2006).from may 2004 to october 2006, a prospective study was carried out in dakar, senegal, to update information about the antimicrobial susceptibility of shigella spp. isolated from stool specimens. among the 165 non-duplicate strains collected, 81 (49%) were identified as shigella flexneri, 75 (45%) as shigella sonnei, 5 (3%) as shigella boydii, and 4 (2%) as shigella dysenteriae. disk diffusion testing revealed that the majority of isolates were resistant to sulphonamides, trimethoprim-sulfamethox ...200818653976
the c-terminus of ipac is required for effector activities related to shigella invasion of host cells.invasion plasmid antigen c (ipac) is secreted by the shigella flexneri type iii secretion system (ttss) as an essential trigger of epithelial cell invasion. at the molecular level, ipac possesses a distinct functional organization. the ipac c-terminal region between amino acids 319 and 345 is predicted to form a coiled-coil structure. such alpha-helical motifs appear to be a recurring structural theme among ttss components. together with ipab, this ipac region is also required for the formation ...200818656530
characterization of soluble complexes of the shigella flexneri type iii secretion system atpase.the conserved atpase of type iii secretion systems is critical to the export of substrates through the apparatus. we present a characterization of the native t3ss atpase, spa47, from the cytoplasm of shigella flexneri, demonstrating it to be in two distinct high-molecular-weight complexes with spa33: mxin and mxik.200818657109
use of mchi encoding immunity to the antimicrobial peptide microcin h47 as a plasmid selection marker in attenuated bacterial live vectors.live attenuated bacterial strains expressing heterologous antigens represent an attractive vaccine development strategy. however, the use of drug resistance genes for the selection of expression plasmids introduced into live vectors poses theoretical health risks. therefore, we developed a novel approach for plasmid selection based on immunity to the antimicrobial peptide microcin h47 (mcch47). two expression plasmids encoding the reporter green fluorescent protein (gfpuv) were constructed; sele ...200818663003
bacterial biosensors for rapid and effective monitoring of biodegradation of organic pollutants in wastewater effluents.significant amounts of toxic substances which are hazardous to animals, plants, microorganisms, and other living organisms including humans are released annually into aquatic and terrestrial environments, mostly from improper wastewater discharges. early detection of such pollutants in wastewater effluents and proper monitoring before their final release into the environment is therefore necessary. in this study, two whole-cell bacterial biosensors were constructed by transforming competent cell ...200818688458
epidemiology of highly endemic multiply antibiotic-resistant shigellosis in children in the peruvian amazon.our goal was to estimate the impact of a shigella vaccine in an area where shigellosis is endemic by characterizing the disease burden and antibiotic-resistance profiles of isolates and by determining the prevalence of shigella flexneri serotypes.200818710884
resistance to fluoroquinolones by the combination of target site mutations and enhanced expression of genes for efflux pumps in shigella flexneri and shigella sonnei strains isolated in korea.among 5938 clinical shigella spp. isolates, two s. flexneri strains were isolated as those resistant to fluoroquinolones based on the mics of the following antibiotics: ciprofloxacin, norfloxacin, ofloxacin, sparfloxacin and levofloxacin. s. flexneri 021787 had three substitutions, one in gyra (ser83leu) and two in parc (ser80ile and arg91gln). s. flexneri 021895 had four substitutions, two in gyra (ser83leu and asp87gly) and two in parc (ser80ile and arg91gln). the increased susceptibility of s ...200818727800
indicator dilution time-activity curves demonstrated by rapid magnetic resonance imaging techniques and paramagnetic contrast agent.indicator dilution time-activity curves are demonstrated using magnetic resonance imaging (mri) as the detector and a paramagnetic contrast agent as the indicator. manganese chloride was injected into a flow phantom. the nuclear magnetic resonance (mr) intensity was measured downstream. several flow rates were used. the observed mr intensity decreased as the paramagnetic indicator passed through the imaging plane. the qualitative changes of the mr intensity decrease varied in accordance with ind ...20082663765
multiple zones in the sequence of calreticulin (crp55, calregulin, hacbp), a major calcium binding er/sr protein.the complete amino acid sequence of crp55, the major 55 kd calcium binding protein of the er lumen, was deduced from the murine cdna nucleotide sequence. this was completed using a novel application of pcr amplification. the mature 399 residue protein encoded is preceded by a 17 amino acid leader sequence and ends in the er signal sequence, kdel. the protein contains no calcium binding motifs of the ef hand type or of the form seen in calelectrin-related proteins. the major region of potential l ...20082583110
multiple zones in the sequence of calreticulin (crp55, calregulin, hacbp), a major calcium binding er/sr protein.the complete amino acid sequence of crp55, the major 55 kd calcium binding protein of the er lumen, was deduced from the murine cdna nucleotide sequence. this was completed using a novel application of pcr amplification. the mature 399 residue protein encoded is preceded by a 17 amino acid leader sequence and ends in the er signal sequence, kdel. the protein contains no calcium binding motifs of the ef hand type or of the form seen in calelectrin-related proteins. the major region of potential l ...20082583110
[minimal monitoring in the operating room]. 20082782684
[minimal monitoring in the operating room]. 20082782684
[what intensity of the incisional pain in the postoperative period?]. 20082576331
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