Publications

characterization and bioactivities of an exopolysaccharide produced by lactobacillus plantarum yw32.	the exopolysaccharide (eps) produced by lactobacillus plantarum yw32 was purified and characterized, and the in vitro bioactivities of the purified eps were also evaluated. the eps had a molecular weight of 1.03×10(5) da, and it consisted of mannose, fructose, galactose and glucose in an approximate molar ratio of 8.2:1:4.1:4.2. microstructural studies of the eps demonstrated a web-like structure composed of compact ropes, and presence of many homogeneous rod-shaped lumps. the eps also showed hi ...	2015	25532782
molecular characterization of multidrug-resistant shigella spp. of food origin.	shigella spp. are the causative agents of food-borne shigellosis, an acute enteric infection. the emergence of multidrug-resistant clinical isolates of shigella presents an increasing challenge for clinicians in the treatment of shigellosis. several studies worldwide have characterized the molecular basis of antibiotic resistance in clinical shigella isolates of human origin, however, to date, no such characterization has been reported for shigella spp. of food origin. in this study, we characte ...	2015	25485847
hpv16l1-attenuated shigella recombinant vaccine induced strong vaginal and systemic immune responses in guinea pig model.	though human papillomavirus (hpv) vaccines based on l1 virus-like particles (vlps) have excellent protective effect against hpv-induced cervical cancer, they are too expensive to be afforded by the developing countries, where most cases of cervical cancer occur. a live bacterial-based vaccine could be an inexpensive alternative. the aim of this study was to evaluate the potential value of live attenuated shigella. flexneri 2a sc602 strain-based hpv16l1 as a high-efficiency, low-cost hpv16l1 muco ...	2014	25483698
outbreak of locally acquired azithromycin-resistant shigella flexneri infection in men who have sex with men.		2015	25474524
structural characterization of a flexible two-domain protein in solution using small angle x-ray scattering and nmr data.	multidomain proteins in which individual domains are connected by linkers often possess inherent interdomain flexibility that significantly complicates their structural characterization in solution using either nuclear magnetic resonance (nmr) spectroscopy or small-angle x-ray scattering (saxs) alone. here, we report a protocol for joint refinement of flexible multidomain protein structures against nmr distance and angular restraints, residual dipolar couplings, and saxs data. the protocol is ba ...	2014	25456817
comparative proteomic analysis of outer membrane vesicles from shigella flexneri under different culture conditions.	the production of outer membrane vesicles (omvs) is a common and regulated process of gram-negative bacteria. nonetheless, the processes of shigella flexneri omv production still remain unclear. s. flexneri is the causative agent of endemic shigellosis in developing countries. the congo red binding of strains is associated with increased infectivity of s. flexneri. therefore, understanding the modulation pattern of omv protein expression induced by congo red will help to elucidate the bacterial ...	2014	25445584
the extant world war 1 dysentery bacillus nctc1: a genomic analysis.	shigellosis (previously bacillary dysentery) was the primary diarrhoeal disease of world war 1, but outbreaks still occur in military operations, and shigellosis causes hundreds of thousands of deaths per year in developing nations. we aimed to generate a high-quality reference genome of the historical shigella flexneri isolate nctc1 and to examine the isolate for resistance to antimicrobials.	2014	25441199
the shigella flexneri ospb effector: an early immunomodulator.	through the action of the type three secretion system (t3ss) shigella flexneri delivers several effectors into host cells to promote cellular invasion, multiplication and to exploit host-cell signaling pathways to modulate the host innate immune response. although much progress has been made in the understanding of many type iii effectors, the molecular and cellular mechanism of the ospb effector is still poorly characterized. in this study we present new evidence that better elucidates the role ...	2015	25434600
the shigella flexneri type 3 secretion system is required for tyrosine kinase-dependent protrusion resolution, and vacuole escape during bacterial dissemination.	shigella flexneri is a human pathogen that triggers its own entry into intestinal cells and escapes primary vacuoles to gain access to the cytosolic compartment. as cytosolic and motile bacteria encounter the cell cortex, they spread from cell to cell through formation of membrane protrusions that resolve into secondary vacuoles in adjacent cells. here, we examined the roles of the type 3 secretion system (t3ss) in s. flexneri dissemination in ht-29 intestinal cells infected with the serotype 2a ...	2014	25405985
update on molecular epidemiology of shigella infection.	shigella spp. are important etiologic agents of diarrhea worldwide. this review summarizes the recent findings on the epidemiology, diagnosis, virulence genes, and pathobiology of shigella infection.	2015	25394237
time-resolved assembly of a nucleoprotein complex between shigella flexneri virf promoter and its transcriptional repressor h-ns.	the virf gene of shigella, responsible for triggering the virulence cascade in this pathogenic bacterium, is transcriptionally repressed by the nucleoid-associated protein h-ns. the primary binding sites of h-ns within the promoter region of virf have been detected here by footprinting experiments in the presence of h-ns or its monomeric dna-binding domain (h-nsctd), which displays the same specificity as intact h-ns. of the 14 short dna fragments identified, 10 overlap sequences similar to the ...	2014	25389261
analysis on the interaction domain of virg and apyrase by pull-down assay.	virg is outer membrane protein of shigella and affects the spread of shigella. recently it has been reported that apyrase influences the location of virg, although the underlying mechanism remains poorly understood. the site of interaction between apyrase and virg is the focus of our research. first we constructed recombinant plasmid phis-phon2 and ps-(v1-1102, v53-758, v759-1102, v53-319, v320-507, v507-758) by denaturation-renaturation, the phon2:kan mutant of shigella flexneri 5a m90t by a mo ...	2014	25379645
a shigella flexneri virulence plasmid encoded factor controls production of outer membrane vesicles.	shigella spp. use a repertoire of virulence plasmid-encoded factors to cause shigellosis. these include components of a type iii secretion apparatus (t3sa) that is required for invasion of epithelial cells and many genes of unknown function. we constructed an array of 99 deletion mutants comprising all genes encoded by the virulence plasmid (excluding those known to be required for plasmid maintenance) of shigella flexneri. we screened these mutants for their ability to bind the dye congo red: a ...	2014	25378474
genome assembly of shigella flexneri atcc 12022, a quality control reference strain.	shigella flexneri causes shigellosis, severe and potentially life-threatening diarrhea, and accounts for 18% of shigellosis cases in the united states. here, we present the 4.51-mbp genome assembly of s. flexneri atcc 12022, a quality control and reference strain, in 10 scaffolds.	2014	25359907
convergent evolution in the assembly of polyubiquitin degradation signals by the shigella flexneri ipah9.8 ligase.	the human pathogen shigella flexneri subverts host function and defenses by deploying a cohort of effector proteins via a type iii secretion system. the ipah family of 10 such effectors mimics ubiquitin ligases but bears no sequence or structural homology to their eukaryotic counterpoints. using rates of (125)i-polyubiquitin chain formation as a functional read out, ipah9.8 displays v-type positive cooperativity with respect to varying concentrations of its ubc5b∼(125)i-ubiquitin thioester co-su ...	2014	25342744
genetic and structural identification of an o-acyltransferase gene (oacc) responsible for the 3/4-o-acetylation on rhamnose iii in shigella flexneri serotype 6.	o-antigen (o-polysaccharide) of the lipopolysaccharide is a highly variable cell component of the outer membrane in shigella flexneri. it defines the serospecificity and plays an important role in the pathogenesis of shigellosis. there are two distinct o-antigen forms for the 19 serotypes of s. flexneri: one for serotypes 1-5, x, y, 7 (and their subtypes), and the other for serotype 6. although having different basal o-polysaccharide structures, the two forms share a common disaccharide fragment ...	2014	25327486
mutational analysis of the shigella flexneri o-antigen polymerase wzy: identification of wzz-dependent wzy mutants.	the o-antigen (oag) component of lipopolysaccharide (lps) is a major virulence determinant of shigella flexneri and is synthesized by the o-antigen polymerase, wzysf. oag chain length is regulated by chromosomally encoded wzzsf and phs-2 plasmid-encoded wzzphs2. to identify functionally important amino acid residues in wzysf, random mutagenesis was performed on the wzysf gene in a pwaldo-tev-gfp plasmid, followed by screening with colicin e2. analysis of the lps conferred by mutated wzysf protei ...	2014	25313393
transcriptional adaptation of shigella flexneri during adherence to epithelial cells.	shigella adhesion to host cells is a transitional stage from an extracellular to an intracellular environment. however, the dynamic adaptations of shigella during adhesion are poorly understood. to address this, we performed the first transcriptome analysis of shigella flexneri 2457t during adhesion. a total of 1,757 genes were differentially regulated (>twofold). the majority of plasmid-borne ipa-mxi-spa locus genes were downregulated, indicating these virulence genes were strictly regulated af ...	2015	25291620
outer membrane protein a (ompa) of shigella flexneri 2a induces tlr2-mediated activation of b cells: involvement of protein tyrosine kinase, erk and nf-κb.	b cells are critically important in combating bacterial infections and their differentiation into plasma cells and memory cells aids bacterial clearance and long-lasting immunity conferred by essentially all vaccines. outer membrane protein a (ompa) of shigella flexneri 2a has been demonstrated to induce the production of igg and iga in vivo following immunization of mice through intranasal route, but the direct involvement of b cells in ompa-mediated immune regulation was not determined. conseq ...	2014	25286253
synthesis, structural characterization, and evaluation of the biological properties of heteroleptic palladium(ii) complexes.	five heteroleptic palladium(ii) complexes of the general formula pd(pr3)(tu)cl2, where pr3 = triphenylphosphine (1), diphenyl-o-tolylphosphine (2), diphenyl-p-tolylphosphine (3), diphenyl-t-butylphosphine (4), and diphenyl-o-methoxyphenylphosphine (5), and tu = 1,3-bis(2-methoxyphenyl) thiourea. they all have been synthesized and characterized by various spectroscopic techniques (elemental analysis, ftir, and (1)h nmr and the ligand 1,3-bis(2-methoxyphenyl) thiourea was synthesized by single cry ...	2014	25276113
clinical isolates of shiga toxin 1a-producing shigella flexneri with an epidemiological link to recent travel to hispañiola.	shiga toxins (stx) are cytotoxins involved in severe human intestinal disease. these toxins are commonly found in shigella dysenteriae serotype 1 and shiga-toxin-producing escherichia coli; however, the toxin genes have been found in other shigella species. we identified 26 shigella flexneri serotype 2 strains isolated by public health laboratories in the united states during 2001-2013, which encode the shiga toxin 1a gene (stx1a). these strains produced and released stx1a as measured by cytotox ...	2014	25271406
high-resolution structure of the shigella type-iii secretion needle by solid-state nmr and cryo-electron microscopy.	we introduce a general hybrid approach for determining the structures of supramolecular assemblies. cryo-electron microscopy (cryo-em) data define the overall envelope of the assembly and rigid-body orientation of the subunits while solid-state nuclear magnetic resonance (ssnmr) chemical shifts and distance constraints define the local secondary structure, protein fold and inter-subunit interactions. finally, rosetta structure calculations provide a general framework to integrate the different s ...	2014	25264107
molecular characterization of serologically atypical provisional serovars of shigella isolates from kolkata, india.	during 2000-2004, 13 shigella strains that were untypable by commercially available antisera were isolated from children <5 years of age with acute diarrhoea in kolkata. these strains were subsequently identified as shigella dysenteriae provisional serovar 204/96 (n = 3), shigella dysenteriae provisional serovar e23507 (n = 1), shigella dysenteriae provisional serovar i9809-73 (n = 1), shigella dysenteriae provisional serovar 93-119 (n = 1), shigella flexneri provisional serovar 88-893 (n = 6) a ...	2014	25261061
isolation and development of bioluminescent reporter phages for bacterial dysentery.	shigellosis is a significant cause of morbidity and mortality worldwide, most notably amongst children. moreover, there is a global increase in the occurrence of multidrug-resistant isolates, including the epidemic and pandemic shigella dysenteriae type 1 strain. we developed a bioluminescent reporter phage assay to facilitate detection and simultaneously determine antibiotic susceptibility. a shigella flexneri phage (shfl25875) was isolated from environmental wastewater and characterized by dna ...	2015	25252629
profile of volatile components of hydrodistilled and extracted leaves of jacaranda acutifolia and their antimicrobial activity against foodborne pathogens.	volatile constituents of the essential oil and n-hexane extract of jacaranda acutifolia humb. and bonpl. (bignoniaceae) leaves were determined, and their antimicrobial activities were investigated using an agar diffusion method. the minimum inhibitory concentrations (mic) were determined and compared with those of standard antibiotics (penicillin, gentamicin and nystatin). the chemical composition of the oils was analyzed by capillary gas chromatography (glc-fid) and gas chromatography-mass spec ...	2014	25230515
[shigella flexneri bacteremia, one adult case and review of the literature].		2014	25229381
use of shigella flexneri to study autophagy-cytoskeleton interactions.	shigella flexneri is an intracellular pathogen that can escape from phagosomes to reach the cytosol, and polymerize the host actin cytoskeleton to promote its motility and dissemination. new work has shown that proteins involved in actin-based motility are also linked to autophagy, an intracellular degradation process crucial for cell autonomous immunity. strikingly, host cells may prevent actin-based motility of s. flexneri by compartmentalizing bacteria inside 'septin cages' and targeting them ...	2014	25226510
shigella flexneri targets the hp1γ subcode through the phosphothreonine lyase ospf.	hp1 proteins are transcriptional regulators that, like histones, are targets for post-translational modifications defining an hp1-mediated subcode. hp1γ has multiple phosphorylation sites, including serine 83 (s83) that marks it to sites of active transcription. in a guinea pig model for shigella enterocolitis, we observed that the defective type iii secretion mxid shigella flexneri strain caused more hp1γ phosphorylation in the colon than the wild-type strain. shigella interferes with hp1 phosp ...	2014	25216677
an optofluidic imaging system to measure the biophysical signature of single waterborne bacteria.	in this paper, for the first time, an on-chip optofluidic imaging system is innovated to measure the biophysical signatures of single waterborne bacteria, including both their refractive indices and morphologies (size and shape), based on immersion refractometry. the key features of the proposed optofluidic imaging platform include (1) multiple sites for single-bacterium trapping, which enable parallel measurements to achieve higher throughput, and (2) a chaotic micromixer, which enables efficie ...	2014	25205636
evaluation of the ability of four esbl-screening media to detect esbl-producing salmonella and shigella.	the aim of this study was to compare the ability of four commercially available media for screening extended-spectrum beta-lactamase (esbl) to detect and identify esbl-producing salmonella and shigella in fecal samples. a total of 71 salmonella- and 21 shigella-isolates producing esbl(a) and/or ampc, were received at norwegian institute of public health between 2005 and 2012. the 92 isolates were mixed with fecal specimens and tested on four esbl screening media; chromid esbl (biomèrieux), brill ...	2014	25204319
detection and analysis of crisprs of shigella.	the recently discovered crisprs (clustered regularly interspaced short palindromic repeats) and cas (crispr-associated) proteins are a novel genetic barrier that limits horizontal gene transfer in prokaryotes and the crispr loci provide a historical view of the exposure of prokaryotes to a variety of foreign genetic elements. the aim of study was to investigate the occurrence and distribution of the crisprs in shigella. a collection of 61 strains of shigella were screened for the existence of cr ...	2015	25199561
detection of salmonella spp. by a loop-mediated isothermal amplification (lamp) method targeting bcfd gene.	in this study, we developed and validated a loop-mediated isothermal amplification (lamp) assay for salmonella detection targeting bcfd gene, a conserved fimbrial operon gene existing in salmonella. the salmonella lamp assay we developed successfully amplified 44 salmonella strains (14 standard strains and 30 clinical isolates), but none of 9 non-salmonella standard strains (proteus mirabilis, listeria monocytogenes, escherichia coli, pseudomonas aeruginosa, shigella flexneri, shigella sonnei, k ...	2014	25199410
shigella flexneri infection in caenorhabditis elegans: cytopathological examination and identification of host responses.	the gram-negative bacterium shigella flexneri is the causative agent of shigellosis, a diarrhoeal disease also known as bacillary dysentery. s. flexneri infects the colonic and rectal epithelia of its primate host and induces a cascade of inflammatory responses that culminates in the destruction of the host intestinal lining. molecular characterization of host-pathogen interactions in this infection has been challenging due to the host specificity of s. flexneri strains, as it strictly infects h ...	2014	25187942
effect of sodium chloride on the survival of shigella flexneri in acidified laboratory media and cucumber puree.	this study investigated the effects of sodium chloride (nacl) and various acids, alone or in combination, on shigella flexneri growth in laboratory medium and cucumber puree.	2014	25178712
serotype-conversion in shigella flexneri: identification of a novel bacteriophage, sf101, from a serotype 7a strain.	shigella flexneri is the major cause of bacillary dysentery in the developing countries. the lipopolysaccharide (lps) o-antigen of s. flexneri plays an important role in its pathogenesis and also divides s. flexneri into 19 serotypes. all the serotypes with an exception for serotype 6 share a common o-antigen backbone comprising of n-acetylglucosamine and three rhamnose residues. different serotypes result from modification of the basic backbone conferred by phage-encoded glucosyltransferase and ...	2014	25174528
proteomes of pathogenic escherichia coli/shigella group surveyed in their host environments.	proteomic studies on shigella dysenteriae, shigella flexneri, enterohemorrhagic escherichia coli and uropathogenic e. coli (upec) are reviewed. upec causes infections in the urogenital tract, whereas the other species colonize and, to varying degrees, invade the intestinal tract. type iii secretion systems used to breach the mucosal barrier by the intestinal pathogens revealed distinct expression patterns in different host environments. dynamic adaptations to changes in nutrient availability and ...	2014	25163594
linear synthesis of the branched pentasaccharide repeats of o-antigens from shigella flexneri 1a and 1b demonstrating the major steric hindrance associated with type-specific glucosylation.	shigella flexneri serotypes 1b and 1a are gram-negative enteroinvasive bacteria causing shigellosis in humans. the o-antigen from s. flexneri 1b is a { → 2)-[3ac/4ac]-α-l-rhap-(1 → 2)-α-l-rhap-(1 → 3)-[2ac]-α-l-rhap-(1 → 3)-[α-d-glcp-(1 → 4)]-β-d-glcpnac-(1 → }n branched polysaccharide ({(ac)ab(ac)c(e)d}n). it is identical to that from s. flexneri 1a, except for the 2c-acetate. a concise synthesis of the disaccharide ed, trisaccharides (ac)c(e)d and c(e)d, tetrasaccharides b(ac)c(e)d and bc(e)d, ...	2014	25141906
enhanced antibacterial and anti-biofilm activities of silver nanoparticles against gram-negative and gram-positive bacteria.	silver nanoparticles (agnps) have been used as antibacterial, antifungal, antiviral, anti-inflammtory, and antiangiogenic due to its unique properties such as physical, chemical, and biological properties. the present study was aimed to investigate antibacterial and anti-biofilm activities of silver nanoparticles alone and in combination with conventional antibiotics against various human pathogenic bacteria. here, we show that a simple, reliable, cost effective and green method for the synthesi ...	2014	25136281
quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone-water extracts of libidibia ferrea, parapiptadenia rigida and psidium guajava.	vast numbers of plant species from northeastern brazil have not yet been phytochemically or biologically evaluated.	2014	25124277
systematic analysis of bacterial effector-postsynaptic density 95/disc large/zonula occludens-1 (pdz) domain interactions demonstrates shigella ospe protein promotes protein kinase c activation via pdlim proteins.	diseases caused by many gram-negative bacterial pathogens depend on the activities of bacterial effector proteins that are delivered into eukaryotic cells via specialized secretion systems. effector protein function largely depends on specific subcellular targeting and specific interactions with cellular ligands. pdz domains are common domains that serve to provide specificity in protein-protein interactions in eukaryotic systems. we show that putative pdz-binding motifs are significantly enrich ...	2014	25124035
multidrug resistant shigella flexneri : a rare case of septicemia in an infant.	shigellosis is still an important public health problem in developing and under-developed countries. it may lead to rare but potentially fatal various extra intestinal complications like septicemia, involvement of cns, urinary tract and liver especially in young malnourished children. the disease is difficult to prevent as only few bacteria are required for causing infection and there is increasing infection with multi drug resistant strains. a 6-month-old infant developed septicemia caused by m ...	2014	25120984
the serine/threonine kinase stk11 promotes shigella flexneri dissemination through establishment of cell-cell contacts competent for tyrosine kinase signaling.	shigella flexneri is an intracellular pathogen that disseminates in the intestinal epithelium by displaying actin-based motility. we found that although s. flexneri displayed comparable actin-based motilities in the cytosols of hela229 and ht-29 epithelial cell lines, the overall dissemination process was much more efficient in ht-29 cells. time-lapse microscopy demonstrated that as motile bacteria reached the cell cortex in ht-29 cells, they formed membrane protrusions that resolved into vacuol ...	2014	25114112
serotype-converting bacteriophage sfii encodes an acyltransferase protein that mediates 6-o-acetylation of glcnac in shigella flexneri o-antigens, conferring on the host a novel o-antigen epitope.	shigella flexneri o-antigen is an important and highly variable cell component presented on the outer leaflet of the outer membrane. most shigella flexneri bacteria share an o-antigen backbone composed of →2)-α-l-rhap(iii)-(1→2)-α-l-rhap(ii)-(1→3)-α-l-rhap(i)-(1→3)-β-d-glcpnac-(1→ repeats, which can be modified by adding various chemical groups to different sugars, giving rise to diverse o-antigen structures and, correspondingly, to various serotypes. the known modifications include glucosylatio ...	2014	25112477
the immune response of two microbial antigens delivered intradermally, sublingually, or the combination thereof.	a key consideration to produce a successful vaccine is the choice of appropriate vaccination route. though most vaccines are administered parenterally, this route is not effective in producing a robust mucosal or cell-mediated response. intradermal and sublingual vaccinations have been explored recently as potential needle-free immunization strategies. we explored intradermal and sublingual routes as well as the combination of the two routes in eliciting both systemic and mucosal immune response ...	2014	25111827
shigella: a highly virulent and elusive pathogen.	despite a significant decrease in shigella-related mortality, shigellosis continues to carry a significant burden of disease worldwide, particularly in asia and africa. shigella is a highly virulent pathogen comprised of four major species with numerous subtypes. shigella dysenteriae and shigella flexneri infections are predominant in resource-limited settings. clinical presentations range from mild watery diarrhea to severe dysentery with systemic complications such as electrolyte imbalance, se ...	2014	25110633
influence of oligomerization state on the structural properties of invasion plasmid antigen b from shigella flexneri in the presence and absence of phospholipid membranes.	shigella flexneri causes bacillary dysentery, an important cause of mortality among children in the developing world. shigella secretes effector proteins via its type iii secretion system (t3ss) to promote bacterial uptake into human colonic epithelial cells. the t3ss basal body spans the bacterial cell envelope anchoring a surface-exposed needle. a pentamer of invasion plasmid antigen d lies at the nascent needle tip and invasion plasmid antigen b (ipab) is recruited into the needle tip complex ...	2014	25103195
sumoylation controls host anti-bacterial response to the gut invasive pathogen shigella flexneri.	shigella flexneri, the etiological agent of bacillary dysentery, invades the human colonic epithelium and causes its massive inflammatory destruction. little is known about the post-translational modifications implicated in regulating the host defense pathway against shigella. here, we show that sumo-2 impairs shigella invasion of epithelial cells in vitro. using mice haploinsufficient for the sumo e2 enzyme, we found that sumoylation regulates intestinal permeability and is required to restrict ...	2014	25097252
bid-dependent release of mitochondrial smac dampens xiap-mediated immunity against shigella.	the x-linked inhibitor of apoptosis protein (xiap) is a potent caspase inhibitor, best known for its anti-apoptotic function in cancer. during apoptosis, xiap is antagonized by smac, which is released from the mitochondria upon caspase-mediated activation of bid. recent studies suggest that xiap is involved in immune signaling. here, we explore xiap as an important mediator of an immune response against the enteroinvasive bacterium shigella flexneri, both in vitro and in vivo. our data demonstra ...	2014	25056906
draft genome sequence of a new shigella flexneri subserotype, 4s bj10610.	shigella flexneri is of great concern in the prevalence of shigellosis and resistance to many antibiotics in developing countries. here, we report the draft genome sequence of a new s. flexneri subserotype, 4s bj10610, isolated from the stool specimens of a patient in beijing, china.	2014	25035336
fluoroquinolone resistance mechanisms of shigella flexneri isolated in bangladesh.	to investigate the prevalence and mechanisms of fluoroquinolone resistance in shigella species isolated in bangladesh and to compare with similar strains isolated in china.	2014	25028972
middle-down mass spectrometry enables characterization of branched ubiquitin chains.	protein ubiquitylation, one of the most prevalent post-translational modifications in eukaryotes, is involved in regulating nearly every cellular signaling pathway. the vast functional range of ubiquitylation has largely been attributed to the formation of a diverse array of polymeric ubiquitin (polyub) chains. methods that enable the characterization of these diverse chains are necessary to fully understand how differences in structure relate to function. here, we describe a method for the dete ...	2014	25023374
modulation of endotoxicity of shigella generalized modules for membrane antigens (gmma) by genetic lipid a modifications: relative activation of tlr4 and tlr2 pathways in different mutants.	outer membrane particles from gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. we previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (gmma), from shigella. as gmma are derived from the outer membrane, they contain immunostimulatory components, especially lipopolysaccharide (lps). we examined ways of reducing their reactogenicity by modifying lip ...	2014	25023285
transfer patterns of integron-associated and antibiotic resistance genes in s. flexneri during different time intervals in tianjin, china.	shigella is one of the common genera of pathogens responsible for bacterial diarrhoea in humans. according to world health organisation (who), 800,000-1,700,000 patients in china were infected with shigella spp. in 2000, and shigella flexneri is the most common serotype (86%).	2014	25008817
widespread antibiotic resistance of diarrheagenic escherichia coli and shigella species.	antibiotic resistance of enteric pathogens particularly shigella species, is a critical world-wide problem and monitoring their resistant pattern is essential, because the choice of antibiotics is absolutely dependent on regional antibiotic susceptibility patterns. during summer 2013, an unusual increase in number of diarrheal diseases was noticed in isfahan, a central province of iran. therefore, the antibiotic resistance of diarrheagenic escherichia coli and shigella species isolated were eval ...	2014	25002896
structural basis for lipopolysaccharide insertion in the bacterial outer membrane.	one of the fundamental properties of biological membranes is the asymmetric distribution of membrane lipids. in gram-negative bacteria, the outer leaflet of the outer membrane is composed predominantly of lipopolysaccharides (lps). the export of lps requires seven essential lipopolysaccharide transport (lpt) proteins to move lps from the inner membrane, through the periplasm to the surface. of the seven lpt proteins, the lptd-lpte complex is responsible for inserting lps into the external leafle ...	2014	24990751
myosin iia is essential for shigella flexneri cell-to-cell spread.	a key feature of shigella pathogenesis is the ability to spread from cell-to-cell post-invasion. this is dependent on the bacteria's ability to initiate de novo f-actin tail polymerisation, followed by protrusion formation, uptake of bacteria-containing protrusion and finally, lysis of the double membrane vacuole in the adjacent cell. in epithelial cells, cytoskeletal tension is maintained by the actin-myosin ii networks. in this study, the role of myosin ii and its specific kinase, myosin light ...	2014	24989342
shigella reroutes host cell central metabolism to obtain high-flux nutrient supply for vigorous intracellular growth.	shigella flexneri proliferate in infected human epithelial cells at exceptionally high rates. this vigorous growth has important consequences for rapid progression to life-threatening bloody diarrhea, but the underlying metabolic mechanisms remain poorly understood. here, we used metabolomics, proteomics, and genetic experiments to determine host and shigella metabolism during infection in a cell culture model. the data suggest that infected host cells maintain largely normal fluxes through glyc ...	2014	24958876
reconstituted human polyclonal plasma-derived secretory-like igm and iga maintain the barrier function of epithelial cells infected with an enteropathogen.	intravenous administration of polyclonal and monoclonal antibodies has proven to be a clinically valid approach in the treatment, or at least relief, of many acute and chronic pathologies, such as infection, immunodeficiency, and a broad range of autoimmune conditions. plasma-derived igg or recombinant igg are most frequently used for intravenous or subcutaneous administration, whereas a few igm-based products are available as well. we have established recently that secretory-like iga and igm ca ...	2014	24951593
atypical haemolytic uraemic syndrome associated with a cd46 mutation triggered by shigella flexneri.	we present a case of haemolytic uraemic syndrome (hus) triggered by shigella flexneri. of the shigella species, only s. dysenteriae type 1 is said to produce shiga toxin and consequently cause hus. investigation of the complement system in this patient revealed a cd46 mutation. in individuals with mutations in complement genes incomplete penetrance of atypical hus (ahus) is seen, suggesting that a trigger, such as infection, is required for disease to manifest. in an era of complement modulatory ...	2014	24944786
elongation factor p and modifying enzyme poxa are necessary for virulence of shigella flexneri.	elongation factor p (ef-p) is a universally conserved bacterial translation factor. in many bacteria, ef-p is posttranslationally modified by poxa, which covalently attaches a β-lysine to a conserved lysine residue of ef-p. here we show that both ef-p and poxa are necessary for virulence of the human diarrheal pathogen shigella flexneri. loss of either ef-p or poxa leads to an impaired ability of s. flexneri to invade epithelial cells and form plaques in an epithelial cell monolayer. proteomic a ...	2014	24935977
biophysical characterization of the type iii secretion tip proteins and the tip proteins attached to bacterium-like particles.	bacterium-like particles (blps), derived from lactococcus lactis, offer a self-adjuvanting delivery vehicle for subunit protein vaccines. proteins can be specifically loaded onto the blps via a peptidoglycan anchoring (pa) domain. in this study, the tip proteins ipad, sipd, and lcrv belonging to type iii secretion systems of shigella flexneri, salmonella enterica, and yersinia enterocolitica, respectively, were fused to the pa and loaded onto the blps. herein, we biophysically characterized thes ...	2015	24916512
probiotic assessment of enterococcus durans 6hl and lactococcus lactis 2hl isolated from vaginal microflora.	forty-five lactic acid bacteria (lab) were isolated from the vaginal specimens of healthy fertile women, and the identities of the bacteria were confirmed by sequencing of their 16s rdna genes. among these bacteria, only four isolates were able to resist and survive in low ph, bile salts and simulated in vitro digestion conditions. lactococcus lactis 2hl, enterococcus durans 6hl, lactobacillus acidophilus 36yl and lactobacillus plantarum 5bl showed the best resistance to these conditions. these ...	2014	24913559
rna-seq analysis of the influence of anaerobiosis and fnr on shigella flexneri.	shigella flexneri is an important human pathogen that has to adapt to the anaerobic environment in the gastrointestinal tract to cause dysentery. to define the influence of anaerobiosis on the virulence of shigella, we performed deep rna sequencing to identify transcriptomic differences that are induced by anaerobiosis and modulated by the anaerobic fumarate and nitrate reduction regulator, fnr.	2014	24907032
travel-related shigellosis in quebec, canada: an analysis of risk factors.	travel-related shigellosis is not well documented in canada although it is frequently acquired abroad and can cause severe disease.	2015	24889090
phyllanthus wightianus müll. arg.: a potential source for natural antimicrobial agents.	phyllanthus wightianus belongs to euphorbiaceae family having ethnobotanical importance. the present study deals with validating the antimicrobial potential of solvent leaf extracts of p. wightianus. 11 human bacterial pathogens (bacillus subtilis, streptococcus pneumoniae, staphylococcus epidermidis, proteus vulgaris, pseudomonas aeruginosa, klebsiella pneumoniae, salmonella typhimurium, escherichia coli, shigella flexneri, proteus vulgaris, and serratia marcescens) and 4 fungal pathogens (cand ...	2014	24883301
cold-active antibacterial and antifungal activities and antibiotic resistance of bacteria isolated from an alpine hydrocarbon-contaminated industrial site.	selection pressure in hydrocarbon-contaminated soils may lead not only to increased microbial resistance to antibiotics, but also to increased capacity of the soil indigenous population to produce antimicrobial compounds. therefore, we studied the antibiotic resistance pattern and antibacterial and/or antifungal activities of 47 bacterial strains isolated from an industrial alpine site heavily polluted with petroleum hydrocarbons. resistance to penicillin was more widespread (49%) than resistanc ...	2014	24880083
biosynthesis of silver nanoparticles using acacia leucophloea extract and their antibacterial activity.	the immense potential of nanobiotechnology makes it an intensely researched field in modern medicine. green nanomaterial synthesis techniques for medicinal applications are desired because of their biocompatibility and lack of toxic byproducts. we report the toxic byproducts free phytosynthesis of stable silver nanoparticles (agnps) using the bark extract of the traditional medicinal plant acacia leucophloea (fabaceae). visual observation, ultraviolet-visible spectroscopy, and transmission elect ...	2014	24876776
shigella flexneri inhibits intestinal inflammation by modulation of host sphingosine-1-phosphate in mice.	infection with invasive shigella species results in intestinal inflammation in humans but no symptoms in adult mice. to investigate why adult mice are resistant to invasive shigellae, 6~8-week-old mice were infected orally with s. flexneri 5a. shigellae successfully colonized the small and large intestines. mild cell death was seen but no inflammation. the infected bacteria were cleared 24 hours later. microarray analysis of infected intestinal tissue showed that several genes that are involved ...	2014	24851099
non-stoichiometric o-acetylation of shigella flexneri 2a o-specific polysaccharide: synthesis and antigenicity.	synthetic functional mimics of the o-antigen from shigella flexneri 2a are seen as promising vaccine components against endemic shigellosis. herein, the influence of the polysaccharide non-stoichiometric di-o-acetylation on antigenicity is addressed for the first time. three decasaccharides, representing relevant internal mono- and di-o-acetylation profiles of the o-antigen, were synthesized from a pivotal protected decasaccharide designed to tailor late stage site-selective o-acetylation. the l ...	2014	24836582
antibacterial and cytotoxic activity of ethanol extract of mikania cordata (burm.f.) b.l. robinson leaves.	the purpose of the present study to examine the antibacterial and cytotoxic properties of ethanol extract of leaves of mikania cordata (burm.f.) b.l. robinson. to determine antibacterial activities, the extract was tested against four gram positive and six gram negative bacteria at three concentrations (500, 800, 1000 μg disc(-1)) through disc diffusion method. the extract showed moderate antibacterial actions and that was increased by increasing the concentration of the sample. the maximum anti ...	2011	24826008
ntrbc and nac contribute to efficient shigella flexneri intracellular replication.	shigella flexneri two-component regulatory systems (tcrs) are responsible for sensing changes in environmental conditions and regulating gene expression accordingly. we examined 12 tcrs that were previously uncharacterized for potential roles in s. flexneri growth within the eukaryotic intracellular environment. we demonstrate that the tcrs evgsa, ntrbc, and rstba systems are required for wild-type plaque formation in cultured epithelial cells. the phenotype of the ntrbc mutant depended in part ...	2014	24794563
decay of genetic markers for fecal bacterial indicators and pathogens in sand from lake superior.	beach sands impact water quality and pathogen loads, however, the comparative decay of the fecal indicator bacteria (fib) enterococcus spp. and escherichia coli, and pathogens in freshwater sand have not been examined. in this study, freshwater sand microcosms were inoculated with sewage and pure cultures of bacterial pathogens to compare relative decay rates. the abundance of culturable enterococcus spp. and e. coli, genetic markers for enterococcus spp. (entero1), total bacteroides (allbac), a ...	2014	24793108
genetic characterization of shigella spp. isolated from diarrhoeal and asymptomatic children.	phenotypic and genetic characteristics of shigella spp. isolated from diarrhoeal and asymptomatic children aged up to 5 years were analysed in this study. in total, 91 and 17 isolates were identified from diarrhoeal (case) and asymptomatic (control) children, respectively. all the isolates were tested for antimicrobial resistance, the presence of integrons, plasmid-mediated quinolone resistance (pmqr), virulence-associated genes and shigella pathogenicity island (sh-pai). the majority of the shi ...	2014	24790083
the periplasmic enzyme, ansb, of shigella flexneri modulates bacterial adherence to host epithelial cells.	s. flexneri strains, most frequently linked with endemic outbreaks of shigellosis, invade the colonic and rectal epithelium of their host and cause severe tissue damage. here we have attempted to elucidate the contribution of the periplasmic enzyme, l-asparaginase (ansb) to the pathogenesis of s. flexneri. using a reverse genetic approach we found that ansb mutants showed reduced adherence to epithelial cells in vitro and attenuation in two in vivo models of shigellosis, the caenorhabditis elega ...	2014	24762742
dynamin-related protein drp1 and mitochondria are important for shigella flexneri infection.	shigella infection in epithelial cells induces cell death which is accompanied by mitochondrial dysfunction. in this study the role of the mitochondrial fission protein, drp1 during shigella infection in hela cells was examined. significant lactate dehydrogenase (ldh) release was detected in the culture supernatant when hela cells were infected with shigella at a high multiplicity of infection. drp1 inhibition with mdivi-1 and sirna knockdown significantly reduced ldh release. hela cell death wa ...	2014	24755420
role of intracellular carbon metabolism pathways in shigella flexneri virulence.	shigella flexneri, which replicates in the cytoplasm of intestinal epithelial cells, can use the embden-meyerhof-parnas, entner-doudoroff, or pentose phosphate pathway for glycolytic carbon metabolism. to determine which of these pathways is used by intracellular s. flexneri, mutants were constructed and tested in a plaque assay for the ability to invade, replicate intracellularly, and spread to adjacent epithelial cells. mutants blocked in the embden-meyerhof-parnas pathway (pfkab and pykaf mut ...	2014	24733092
murine solid tumours as a novel model to study bacterial biofilm formation in vivo.	bacteria of many species are able to invade and colonize solid tumours in mice. we have focused on salmonella enterica serovar typhimurium. detailed analysis revealed that such tumour-invading salmonella form biofilms, thus providing a versatile in vivo test system for studying bacterial phenotypes and host-pathogen interactions. it appears that biofilm formation by s. typhimurium is induced as a defence against the immune system of the host, and in particular against neutrophils. further, we ex ...	2014	24724621
icsa is a shigella flexneri adhesin regulated by the type iii secretion system and required for pathogenesis.	following contact with the epithelium, the enteric intracellular bacterial pathogen shigella flexneri invades epithelial cells and escapes intracellular phagosomal destruction using its type iii secretion system (t3ss). the bacterium replicates within the host cell cytosol and spreads between cells using actin-based motility, which is mediated by the virulence factor icsa (virg). whereas s. flexneri invasion is well characterized, adhesion mechanisms of the bacterium remain elusive. we found tha ...	2014	24721572
[preparation and identification of the polyclonal antibody against gapdh protein from shigella flexneri m90t].	to prepare mouse polyclonal antibodies against shigella flexneri m90t glyceraldehyde-3-phosphate dehydrogenase (gapdh), and verify its titer and specificity.	2014	24721410
draft genome sequences of the type strains of shigella flexneri held at public health england: comparison of classical phenotypic and novel molecular assays with whole genome sequence.	public health england (phe) holds a collection of shigella flexneri type strains isolated between 1949 and 1972 representing 15 established serotypes and one provisional type, e1037. in this study, the genomes of all 16 phe type strains were sequenced using the illumina hiseq platform. the relationship between core genome phylogeny and serotype was examined.	2014	24684748
probiotic potential of lactobacillus plantarum ld1 isolated from batter of dosa, a south indian fermented food.	lactobacillus plantarum ld1 was isolated from dosa batter and identified by biochemical, physiological and genetic methods. species level identification was done by 16s rdna amplification and sequencing. the probiotic potential of strain ld1 was assessed by different standard parameters. cell surface hydrophobicity was recorded to be 62% with sat value <0.007 m. seventy-eight percent of viable count was found after treatment with simulated gastric juice containing pepsin (ph 2.0). bile salt tole ...	2014	24676724
ompa and ompc are critical host factors for bacteriophage sf6 entry in shigella.	despite being essential for successful infection, the molecular cues involved in host recognition and genome transfer of viruses are not completely understood. bacterial outer membrane proteins a and c co-purify in lipid vesicles with bacteriophage sf6, implicating both outer membrane proteins as potential host receptors. we determined that outer membrane proteins a and c mediate sf6 infection by dramatically increasing its rate and efficiency. we performed a combination of in vivo studies with ...	2014	24673644
serological identification and prevalence of a novel o-antigen epitope linked to 3- and 4-o-acetylated rhamnose iii of lipopolysaccharide in shigella flexneri.	shigella flexneri is the major cause of shigellosis in developing countries. all serotypes except for serotype 6 share an o-antigen backbone composed of a → 2)-α-l-rhap(iii)-(1 → 2)-α-l-rhap(ii)-(1 → 3)-α-l-rhap(i)-(1 → 3)-β-d-glcpnac-(1 → tetrasaccharide repeat. it can be modified by the addition of a glucosyl group to one or more sugar residues and/or an o-acetyl group to rha(i) and/or a phosphoethanolamine to rha(ii) and/or rha(iii). these modifications give rise to type i-, ic-, ii-, iv-, an ...	2014	24671799
tyrosine kinases, drugs, and shigella flexneri dissemination.	shigella flexneri is an enteropathogenic bacterium responsible for approximately 100 million cases of severe dysentery each year. s. flexneri colonization of the human colonic epithelium is supported by direct spread from cell to cell, which relies on actin-based motility. we have recently uncovered that, in intestinal epithelial cells, s. flexneri actin-based motility is regulated by the bruton's tyrosine kinase (btk). consequently, treatment with ibrutinib, a specific btk inhibitor currently u ...	2017	24637806
rabbit monoclonal antibodies directed at the t3ss effector protein yopm identify human pathogenic yersinia isolates.	the yersinia outer protein m (yopm) is a type 3 secretion system (t3ss)-dependent effector protein of yersinia enterocolitica, yersinia pseudotuberculosis and yersinia pestis. although yopm is indispensable for full virulence, its molecular functions still remain largely elusive. recently, we could identify the recombinant yopm (ryopm) protein derived from the y. enterocolitica strain 8081 (jb580) as a cell-penetrating protein, which down-regulates the expression of various pro-inflammatory cyto ...	2014	24636859
new insights into the crosstalk between shigella and t lymphocytes.	subversion of host immune responses is the key infection strategy employed by most, if not all, human pathogens. modulation of the host innate response by pathogens has been vastly documented. yet, especially for bacterial infections, it was only recently that cells of the adaptive immune response were recognized as targets of bacterial weapons such as the type iii secretion system (t3ss) and its effector proteins. in this review, we focus on the recent advances made in the understanding of how ...	2014	24613405
development of an aotus nancymaae model for shigella vaccine immunogenicity and efficacy studies.	several animal models exist to evaluate the immunogenicity and protective efficacy of candidate shigella vaccines. the two most widely used nonprimate models for vaccine development include a murine pulmonary challenge model and a guinea pig keratoconjunctivitis model. nonhuman primate models exhibit clinical features and gross and microscopic colonic lesions that mimic those induced in human shigellosis. challenge models for enterotoxigenic escherichia coli (etec) and campylobacter spp. have be ...	2014	24595138
polar localization of phon2, a periplasmic virulence-associated factor of shigella flexneri, is required for proper icsa exposition at the old bacterial pole.	proper protein localization is critical for bacterial virulence. phon2 is a virulence-associated atp-diphosphohydrolase (apyrase) involved in icsa-mediated actin-based motility of s. flexneri. herein, by analyzing a δphon2 mutant of the s. flexneri strain m90t and by generating phon2::ha fusions, we show that phon2, is a periplasmic protein that strictly localizes at the bacterial poles, with a strong preference for the old pole, the pole where icsa is exposed, and that it is required for proper ...	2014	24587292
synthesis, structures, spectroscopy and antimicrobial properties of complexes of copper(ii) with salicylaldehyde n-substituted thiosemicarbazones and 2,2'-bipyridine or 1,10-phenanthroline.	among the biometals (cu, co, ni-cofactors in many enzymes), copper derivatives of o, n, s-donor salicylaldehyde thiosemicarbazones have received considerable attention owing to their potential biological applications. eight new complexes of salicylaldehyde-n-substituted thiosemicarbazones [5-meo-2-ho-c₆h₄-c(2)(h)n(3)-n(2)h-c(1)(s)-n(1)hr; r = me, h2l(1); et, h₂l(1), ph, h₂l(3), h, h₂l(4)] with copper(ii), namely, [cu(κ(3)-o,n,s-l)( κ(2)-n,n-l')] {(l)(2-) = (l(1))(2-), l' = bipy, 1, phen, 2; (l)( ...	2014	24583354
conformational diversity of o-antigen polysaccharides of the gram-negative bacterium shigella flexneri serotype y.	o-antigen polysaccharides constitute the outer protective layer of most gram-negative bacteria, important for the bacterium's survival and adaption within its host. although important for many functions, the three-dimensional structure of the dense polysaccharide coat remains to be elucidated. in this study, we present a systematic numerical investigation of o-antigen polysaccharide chains of shigella flexneri serotype y composed of one up to four tetrasaccharide repeat units. to bridge the gap ...	2014	24559142
potential novel antibiotics from hts targeting the virulence-regulating transcription factor, virf, from shigella flexneri.	virf is an arac-type transcriptional regulator responsible for activating the transcription of virulence genes required for the intracellular invasion and cell-to-cell spread of shigella flexneri. gene disruption studies have validated virf as a potential target for an anti-virulence therapy to treat shigellosis by determining that virf is necessary for virulence, but not required for bacterial viability. using a bacteria-based, β-galactosidase reporter assay we completed a high-throughput scree ...	2014	24549153
[stability of changed properties of streptomycin-resistant strain of shigella flexneri after prolonged preservation].		2014	24544446
[result of serological studies of standard cultures of shigella flexneri obtained from various museum collections].		1953	24543446
[data on biological characteristics of shigella flexneri].		1953	24543445
a fluorescent reporter reveals on/off regulation of the shigella type iii secretion apparatus during entry and cell-to-cell spread.	numerous pathogenic gram-negative bacteria use a type three secretion apparatus (t3sa) to translocate effector proteins into host cells. detecting and monitoring the t3sa of intracellular bacteria within intact host cells has been challenging. taking advantage of the tight coupling between t3s effector-gene transcription and t3sa activity in shigella flexneri, together with a fast-maturing green fluorescent protein, we developed reporters to monitor t3sa activity in real time. these reporters re ...	2014	24528864
plantaricin ld1: a bacteriocin produced by food isolate of lactobacillus plantarum ld1.	plantaricin ld1, a bacteriocin produced by lactobacillus plantarum ld1, was characterized for biochemical and antimicrobial properties. bacteriocin showed stability at high temperatures (100 °c for 20 min and 121 °c for 15 min under 15 psi pressure), in a ph range of 2.0-8.0 and also in the presence of organic solvents, surfactants and detergents. the crude preparation was not affected by catalase, amylase and lipase but activity was reduced in the presence of pepsin, trypsin and proteinase k sh ...	2014	24522411
oral administration of live shigella vaccine candidates in rhesus monkeys show no evidence of competition for colonization and immunogenicity between different serotypes.	live oral monovalent shigella flexneri 2a vaccine candidates as well as bivalent formulations with shigella sonnei were evaluated in a rhesus monkey model for colonization and immunogenicity. freshly harvested suspensions of s. flexneri 2a vaccine candidates wrsf2g12 and wrsf2g15 as well as s. sonnei vaccine candidate wrss3 were nasogastrically administered to groups of rhesus monkeys, macaca mulatta, either in a monovalent form or when combined with each other. the animals were monitored daily ...	2014	24522159
shigellemia in a post renal transplant patient: a case report and literature review.	shigellemia is a complication of shigellosis that occurs generally in malnourished children. in adults, shigellemia is usually seen in immunocompromised individuals. here we report the first case of shigellemia in a renal transplant patient from india. the patient had history of diarrhea, which was treated symptomatically. subsequently, the patient developed high-grade fever and blood culture was positive for shigella flexneri. recovery was uneventful after the initiation of antimicrobial therap ...	2014	24518636
identification of an o-acyltransferase gene (oacb) that mediates 3- and 4-o-acetylation of rhamnose iii in shigella flexneri o antigens.	o antigen (o polysaccharide) is an important and highly variable cell component present on the surface of cells which defines the serospecificity of gram-negative bacteria. most o antigens of shigella flexneri, a cause of shigellosis, share a backbone composed of →2)-α-l-rhap(iii)-(1→2)-α-l-rhap(ii)-(1→3)-α-l-rhap(i)-(1→3)-β-d-glcpnac-(1→ repeats, which can be modified by adding various substituents, giving rise to 19 serotypes. the known modifications include glucosylation on various sugar resi ...	2014	24509314
multiplex pcr (polymerase chain reaction) assay for detection of e. coli o157:h7, salmonella sp., vibrio cholerae and vibrio parahaemolyticus in spiked shrimps (penaeus monodon).	the coastal aquaculture mainly shrimps constitute major export sector in bangladesh and is increasingly shaped by international trade conditions and by national responses to those stringent quality and safety standards. pcr based validated methods for detection of major bacterial pathogens in shrimp might be very useful tool for ensuring quality and safety standards of exportable shrimps. the objective of this study was to evaluate overall performance (sensitivity and specificity) of the multipl ...	2013	24498789