Publications

TitleAbstractYear(sorted ascending)
Filter
PMID
Filter
synergistic effects of probiotic leuconostoc mesenteroides and bacillus subtilis in malted ragi (eleucine corocana) food for antagonistic activity against v. cholerae and other beneficial properties.finger millet (elucine corocana), locally known as ragi, and probiotics have been recognized for their health benefits. in the present work we describe novel probiotic ragi malt (functional food) that has been prepared using ragi and probiotic leuconostoc mesenteroides (lm) and bacillus subtilis natto (bs), alone and in combination, for antagonistic activity against vibrio cholerae (vc). in vitro studies using pure cultures showed that each probiotic strain (lm or bs) was able to inhibit the pla ...201226396299
insight into the bacterial gut microbiome of the north american moose (alces alces).the work presented here provides the first intensive insight into the bacterial populations in the digestive tract of the north american moose (alces alces). eight free-range moose on natural pasture were sampled, producing eight rumen samples and six colon samples. second generation (g2) phylochips were used to determine the presence of hundreds of operational taxonomic units (otus), representing multiple closely related species/strains (>97% identity), found in the rumen and colon of the moose ...201222992344
hydrogen-bonded networks along and bifurcation of the e-pathway in quinol:fumarate reductase.the e-pathway of transmembrane proton transfer has been demonstrated previously to be essential for catalysis by the diheme-containing quinol:fumarate reductase (qfr) of wolinella succinogenes. two constituents of this pathway, glu-c180 and heme b(d) ring c (b(d)-c-) propionate, have been validated experimentally. here, we identify further constituents of the e-pathway by analysis of molecular dynamics simulations. the redox state of heme groups has a crucial effect on the connectivity patterns ...201222995503
domain organization of the polymerizing mannosyltransferases involved in synthesis of the escherichia coli o8 and o9a lipopolysaccharide o-antigens.the escherichia coli o9a and o8 polymannose o-polysaccharides (o-pss) serve as model systems for the biosynthesis of bacterial polysaccharides by atp-binding cassette transporter-dependent pathways. both o-pss contain a conserved primer-adaptor domain at the reducing terminus and a serotype-specific repeat unit domain. the repeat unit domain is polymerized by the serotype-specific wbda mannosyltransferase. in serotype o9a, wbda is a bifunctional α-(1→2)-, α-(1→3)-mannosyltransferase, and its cou ...201222989876
phase variable o antigen biosynthetic genes control expression of the major protective antigen and bacteriophage receptor in vibrio cholerae o1.the vibrio cholerae lipopolysaccharide o1 antigen is a major target of bacteriophages and the human immune system and is of critical importance for vaccine design. we used an o1-specific lytic bacteriophage as a tool to probe the capacity of v. cholerae to alter its o1 antigen and identified a novel mechanism by which this organism can modulate o antigen expression and exhibit intra-strain heterogeneity. we identified two phase variable genes required for o1 antigen biosynthesis, mana and wbel. ...201223028317
iron and ph-responsive ftrabcd ferrous iron utilization system of bordetella species.a putative operon encoding an uncharacterized ferrous iron transport (ftrabcd) system was previously identified in cdna microarray studies. in growth studies using buffered medium at ph values ranging from ph 6.0 to 7.6, bordetella pertussis and bordetella bronchiseptica ftrabcd system mutants showed dramatic reductions in growth yields under iron-restricted conditions at ph 6.0, but had no growth defects at ph 7.6. supplementation of culture medium with 2 mm ascorbate reductant was inhibitory t ...201222924881
roles for wbtc, wbti, and kdta genes in lipopolysaccharide biosynthesis, protein glycosylation, virulence, and immunogenicity in francisella tularensis2 strain schu s4.using a strategy of gene deletion mutagenesis, we have examined the roles of genes putatively involved in lipopolysaccharide biosynthesis in the virulent facultative intracellular bacterial pathogen, francisella tularensis subspecies tularensis, strain schu s4 in lps biosynthesis, protein glycosylation, virulence and immunogenicity. one mutant, δwbti, did not elaborate a long chain o-polysaccharide (ops), was completely avirulent for mice, and failed to induce a protective immune response agains ...201225152813
campylobacter jejuni induces extra-intestinal immune responses via toll-like-receptor-4 signaling in conventional il-10 deficient mice with chronic colitis.campylobacter jejuni is one of the predominant causes for foodborne bacterial infections worldwide. we investigated whether signaling of c. jejuni-lipoproteins and -lipooligosaccharide via toll-like-receptor (tlr) -2 and -4, respectively, is inducing intestinal and extra-intestinal immune responses following infection of conventional il-10(-/-) mice with chronic colitis. at day 3 following oral infection, il-10(-/-) mice lacking tlr-2 or tlr-4 harbored comparable c. jejuni strain atcc 43431 load ...201224688768
the main aeromonas pathogenic factors.the members of the aeromonas genus are ubiquitous, water-borne bacteria. they have been isolated from marine waters, rivers, lakes, swamps, sediments, chlorine water, water distribution systems, drinking water and residual waters; different types of food, such as meat, fish, seafood, vegetables, and processed foods. aeromonas strains are predominantly pathogenic to poikilothermic animals, and the mesophilic strains are emerging as important pathogens in humans, causing a variety of extraintestin ...201223724321
production of organic acids by probiotic lactobacilli can be used to reduce pathogen load in poultry.probiotic lactobacillus can be used to reduce the colonization of pathogenic bacteria in food animals, and therefore reduce the risk of foodborne illness to consumers. as a model system, we examined the mechanism of protection conferred by lactobacillus species to inhibit c. jejuni growth in vitro and reduce colonization in broiler chickens. possible mechanisms for the reduction of pathogens by lactobacilli include: 1) stimulation of adaptive immunity; 2) alteration of the cecal microbiome; and, ...201222962594
atypical findings of guillain-barré syndrome in children.guillain-barre syndrome (gbs) is an immune-mediated polyneuropathy that occurs mostly after prior infection. the diagnosis of this syndrome is dependent heavily on the history and examination, although cerebrospinal fluid analysis and electrodiagnostic testing usually confirm the diagnosis. this is a retrospective study which was performed to investigate the atypical features of gbs.201224665275
milk-borne infections. an analysis of their potential effect on the milk industry.in developed countries such as the united states of america, foodborne illnesses account for 48 million infections per year. developing countries such as india face greater simultaneous challenges particularly since incorrect processing or storage of dairy products can represent a transmission hazard for a large number of pathogens and can be responsible for outbreaks of brucellosis, listeriosis, tuberculosis, etc. it is important to recognize the types of germs which can be transmitted through ...201224432270
beyond growth: novel functions for bacterial cell wall hydrolases.the peptidoglycan cell wall maintains turgor pressure and cell shape of most bacteria. cell wall hydrolases are essential, together with synthases, for growth and daughter cell separation. recent work in diverse organisms has uncovered new cell wall hydrolases that act autonomously or on neighboring cells to modulate invasion of prey cells, cell shape, innate immune detection, intercellular communication, and competitor lysis. the hydrolases involved in these processes catalyze the cleavage of b ...201222944244
infectious disease outbreaks in competitive sports, 2005-2010.old, evolving, and new infectious agents continually threaten the participation of competitors in sports.201223068588
inhibitors of b-cell receptor signaling for patients with b-cell malignancies.the b-cell receptor (bcr) complex and its associated protein tyrosine kinases play a critical role in the development, proliferation, and survival of normal or malignant b cells. regulated activity of the bcr complex promotes the expansion of selected b cells and the deletion of unwanted or self-reactive ones. compounds that inhibit various components of this pathway, including spleen tyrosine kinase, bruton's tyrosine kinase, and phosphoinositol-3 kinase, have been developed. we summarize the r ...201223006944
hypomorphic glycosyltransferase alleles and recoding at contingency loci influence glycan microheterogeneity in the protein glycosylation system of neisseria species.as more bacterial protein glycosylation systems are identified and characterized, a central question that arises is, what governs the prevalence of particular glycans associated with them? in addition, accumulating evidence shows that bacterial protein glycans can be subject to the phenomenon of microheterogeneity, in which variant glycan structures are found at specific attachment sites of a given glycoprotein. although factors underlying microheterogeneity in reconstituted expression systems h ...201222797763
the role of bacterial enhancer binding proteins as specialized activators of σ54-dependent transcription.bacterial enhancer binding proteins (bebps) are transcriptional activators that assemble as hexameric rings in their active forms and utilize atp hydrolysis to remodel the conformation of rna polymerase containing the alternative sigma factor σ(54). we present a comprehensive and detailed summary of recent advances in our understanding of how these specialized molecular machines function. the review is structured by introducing each of the three domains in turn: the central catalytic domain, the ...201222933558
the campylobacter jejuni dps homologue is important for in vitro biofilm formation and cecal colonization of poultry and may serve as a protective antigen for vaccination.in this work, we investigated the campylobacter jejuni dps (dna binding protein from starved cells) gene for a role in biofilm formation and cecal colonization in poultry. in vitro biofilm formation assays were conducted with stationary-phase cells in cell culture plates under microaerophilic conditions. these studies demonstrated a significant (>50%) reduction in biofilm formation by the c. jejuni dps mutant compared to that by the wild-type strain. studies in poultry also demonstrated the impo ...201222787197
development of anti-infectives using phage display: biological agents against bacteria, viruses, and parasites.the vast majority of anti-infective therapeutics on the market or in development are small molecules; however, there is now a nascent pipeline of biological agents in development. until recently, phage display technologies were used mainly to produce monoclonal antibodies (mabs) targeted against cancer or inflammatory disease targets. patent disputes impeded broad use of these methods and contributed to the dearth of candidates in the clinic during the 1990s. today, however, phage display is rec ...201222664969
comprehensive detection and discrimination of campylobacter species by use of confocal micro-raman spectroscopy and multilocus sequence typing.a novel strategy for the rapid detection and identification of traditional and emerging campylobacter strains based upon raman spectroscopy (532 nm) is presented here. a total of 200 reference strains and clinical isolates of 11 different campylobacter species recovered from infected animals and humans from china and north america were used to establish a global raman spectroscopy-based dendrogram model for campylobacter identification to the species level and cross validated for its feasibility ...201222740711
mutations that impact the enteropathogenic escherichia coli cpx envelope stress response attenuate virulence in galleria mellonella.in this paper, we show that the larvae of the greater wax moth, galleria mellonella, can be used as a model to study enteropathogenic escherichia coli (epec) virulence. g. mellonella larvae are killed after infection with epec type strain e2348/69 but not by an attenuated derivative that expresses diminished levels of the major virulence determinants or by a mutant specifically defective in type iii secretion (t3s). infecting epec inhabit the larval hemocoel only briefly and then become localize ...201222710873
role of rela and spot in burkholderia pseudomallei virulence and immunity.burkholderia pseudomallei is a gram-negative soil bacterium and the causative agent of melioidosis, a disease of humans and animals. it is also listed as a category b bioterrorism threat agent by the u.s. centers for disease control and prevention, and there is currently no melioidosis vaccine available. small modified nucleotides such as the hyperphosphorylated guanosine molecules ppgpp and pppgpp play an important role as signaling molecules in prokaryotes. they mediate a global stress respons ...201222778096
campylobacter jejuni translocation across intestinal epithelial cells is facilitated by ganglioside-like lipooligosaccharide structures.translocation across intestinal epithelial cells is an established pathogenic feature of the zoonotic bacterial species campylobacter jejuni. the number of c. jejuni virulence factors known to be involved in translocation is limited. in the present study, we investigated whether sialylation of c. jejuni lipooligosaccharide (los) structures, generating human nerve ganglioside mimics, is important for intestinal epithelial translocation. we here show that c. jejuni isolates expressing ganglioside- ...201222778098
gastrointestinal lymphomas: morphology, immunophenotype and molecular features.primary gastrointestinal lymphoma comprises 10-15% of all non-hodgkin lymphomas and encompasses 30-40% of the total extranodal lymphomas. approximately 60-75% of cases occur in the stomach, and then the small bowel, ileum, cecum, colon and rectum. lymphoid neoplasms may consist of mature b, t and less commonly extranodal nk/t cells. of these, the two most frequently encountered histologic subtypes are extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (malt lymphoma), where h ...201222943012
dnak dependence of the mycobacterial stress-responsive regulator hspr is mediated through its hydrophobic c-terminal tail.hspr is a repressor known to control expression of heat shock operons in a number of eubacteria. in mycobacteria and in several other actinobacteria, this protein is synthesized from the dnakje-hspr operon. previous investigations revealed that hspr binds to the operon promoter, thereby controlling its expression in an autoregulatory manner. dnak, which is a product of the same operon, further aids this autoregulatory process by stimulating the operator binding activity of hspr. the molecular me ...201222753065
double-stranded endonuclease activity in bacillus halodurans clustered regularly interspaced short palindromic repeats (crispr)-associated cas2 protein.the crispr (clustered regularly interspaced short palindromic repeats) system is a prokaryotic rna-based adaptive immune system against extrachromosomal genetic elements. cas2 is a universally conserved core crispr-associated protein required for the acquisition of new spacers for crispr adaptation. it was previously characterized as an endoribonuclease with preference for single-stranded (ss)rna. here, we show using crystallography, mutagenesis, and isothermal titration calorimetry that the bac ...201222942283
interdisciplinary approaches to understanding disease emergence: the past, present, and future drivers of nipah virus emergence.emerging infectious diseases (eids) pose a significant threat to human health, economic stability, and biodiversity. despite this, the mechanisms underlying disease emergence are still not fully understood, and control measures rely heavily on mitigating the impact of eids after they have emerged. here, we highlight the emergence of a zoonotic henipavirus, nipah virus, to demonstrate the interdisciplinary and macroecological approaches necessary to understand eid emergence. previous work suggest ...201222936052
interdisciplinary approaches to understanding disease emergence: the past, present, and future drivers of nipah virus emergence.emerging infectious diseases (eids) pose a significant threat to human health, economic stability, and biodiversity. despite this, the mechanisms underlying disease emergence are still not fully understood, and control measures rely heavily on mitigating the impact of eids after they have emerged. here, we highlight the emergence of a zoonotic henipavirus, nipah virus, to demonstrate the interdisciplinary and macroecological approaches necessary to understand eid emergence. previous work suggest ...201222936052
porphyromonas gingivalis entry into gingival epithelial cells modulated by fusobacterium nucleatum is dependent on lipid rafts.host cell invasion by a major periodontal pathogen, porphyromonas gingivalis, has been proposed as an important mechanism involved in host-pathogen interactions in periodontal and cardiovascular diseases. the present study sought to gain insight into the underlying mechanism(s) involved in previously demonstrated fusobacterial modulation of host cell invasion by p. gingivalis. an immortalized human gingival cell line ca9-22 was dually infected with p. gingivalis atcc 33277 and fusobacterium nucl ...201223034475
structure of anabolic ornithine carbamoyltransferase from campylobacter jejuni at 2.7 å resolution.anabolic ornithine transcarbamoylase (aotc) catalyzes the reaction between carbamoyl phosphate (cp) and l-ornithine (orn) to form l-citrulline and phosphate in the urea cycle and l-arginine biosynthesis. the crystal structure of unliganded aotc from campylobacter jejuni (cje aotc) was determined at 2.7 å resolution and refined to an r(work) of 20.3% and an r(free) of 24.0%. cje aotc is a trimer that forms a head-to-head pseudohexamer in the asymmetric unit. each monomer is composed of an n-termi ...201222949186
lacto-n-tetraose, fucosylation, and secretor status are highly variable in human milk oligosaccharides from women delivering preterm.breast milk is the ideal nutrition for term infants but must be supplemented to provide adequate growth for most premature infants. human milk oligosaccharides (hmos) are remarkably abundant and diverse in breast milk and yet provide no nutritive value to the infant. hmos appear to have at least two major functions: prebiotic activity (stimulation of the growth of commensal bacteria in the gut) and protection against pathogens. investigations of hmos in milk from women delivering preterm have be ...201222900748
campylobacter jejuni type vi secretion system: roles in adaptation to deoxycholic acid, host cell adherence, invasion, and in vivo colonization.the recently identified type vi secretion system (t6ss) of proteobacteria has been shown to promote pathogenicity, competitive advantage over competing microorganisms, and adaptation to environmental perturbation. by detailed phenotypic characterization of loss-of-function mutants, in silico, in vitro and in vivo analyses, we provide evidence that the enteric pathogen, campylobacter jejuni, possesses a functional t6ss and that the secretion system exerts pleiotropic effects on two crucial proces ...201222952616
guillain-barré syndrome-related campylobacter jejuni in bangladesh: ganglioside mimicry and cross-reactive antibodies.campylobacter jejuni is the predominant antecedent infection in guillain-barré syndrome (gbs). molecular mimicry and cross-reactive immune responses to c. jejuni lipo-oligosaccharides (los) precipitate the development of gbs, although this mechanism has not been established in patients from developing countries. we determined the carbohydrate mimicry between c. jejuni los and gangliosides, and the cross-reactive antibody response in patients with gbs in bangladesh.201222952833
serologic host response to helicobacter pylori and campylobacter jejuni in socially housed rhesus macaques (macaca mulatta). 201222920270
relevance of anxiety in clinical practice of guillain-barre syndrome: a cohort study.illness is often associated with anxiety, but few data exist about the prognostic significance of this phenomenon. to address this issue, we assessed whether patient anxiety is associated with subsequent need for intubation in guillain-barré syndrome (gbs).201222923622
a picomolar transition state analogue inhibitor of mtan as a specific antibiotic for helicobacter pylori.campylobacter and helicobacter species express a 6-amino-6-deoxyfutalosine n-ribosylhydrolase (hpmtan) proposed to function in menaquinone synthesis. but-dadme-imma is a 36 pm transition state analogue of hpmtan, and the crystal structure of the enzyme-inhibitor complex reveals the mechanism of inhibition. but-dadme-imma has a mic(90) value of <8 ng/ml for helicobacter pylori growth but does not cause growth arrest in other common clinical pathogens, thus demonstrating potential as an h. pylori- ...201222891633
beta-adrenergic receptor 1 selective antagonism inhibits norepinephrine-mediated tnf-alpha downregulation in experimental liver cirrhosis.bacterial translocation is a frequent event in cirrhosis leading to an increased inflammatory response. splanchnic adrenergic system hyperactivation has been related with increased bacterial translocation. we aim at evaluating the interacting mechanism between hepatic norepinephrine and inflammation during liver damage in the presence of bacterial-dna.201222916250
yersinia pestis infection and laboratory conditions alter flea-associated bacterial communities.we collected oropsylla montana from rock squirrels, spermophilus varigatus, and infected a subset of collected fleas with yersinia pestis, the etiological agent of plague. we used bar-tagged dna pyrosequencing to characterize bacterial communities of wild, uninfected controls and infected fleas. bacterial communities within y. pestis-infected fleas were substantially more similar to one another than communities within wild or control fleas, suggesting that infection alters the bacterial communit ...201222895162
yersinia pestis infection and laboratory conditions alter flea-associated bacterial communities.we collected oropsylla montana from rock squirrels, spermophilus varigatus, and infected a subset of collected fleas with yersinia pestis, the etiological agent of plague. we used bar-tagged dna pyrosequencing to characterize bacterial communities of wild, uninfected controls and infected fleas. bacterial communities within y. pestis-infected fleas were substantially more similar to one another than communities within wild or control fleas, suggesting that infection alters the bacterial communit ...201222895162
application of orange essential oil as an antistaphylococcal agent in a dressing model.staphylococcus aureus is the pathogen most often and prevalently involved in skin and soft tissue infections. in recent decades outbreaks of methicillin-resistant s. aureus (mrsa) have created major problems for skin therapy, and burn and wound care units. topical antimicrobials are most important component of wound infection therapy. alternative therapies are being sought for treatment of mrsa and one area of interest is the use of essential oils. with the increasing interest in the use and app ...201222894560
guillain-barre syndrome after coronary artery bypass surgery.a case of the guillain-barré syndrome occurring after otherwise uneventful cardiac surgery using cardiopulmonary bypass is presented. though the guillain-barré syndrome has been reported after surgical procedures, there are very few case reports after cardiopulmonary bypass surgery in the literature. the exact pathophysiological cause of the syndrome is still unknown. however, the most widely accepted hypothesis is that the syndrome is the result of an immune-mediated process. cardiac surgery ma ...201222899666
inhibitory effects of 405 nm irradiation on chlamydia trachomatis growth and characterization of the ensuing inflammatory response in hela cells.chlamydia trachomatis is an intracellular bacterium that resides in the conjunctival and reproductive tract mucosae and is responsible for an array of acute and chronic diseases. a percentage of these infections persist even after use of antibiotics, suggesting the need for alternative treatments. previous studies have demonstrated anti-bacterial effects using different wavelengths of visible light at varying energy densities, though only against extracellular bacteria. we investigated the effec ...201222894815
salmonella "sops" up a preferred electron receptor in the inflamed intestine.the microbiota of the mammalian intestinal tract represents a formidable barrier to colonization by pathogens. to overcome this resistance to colonization, bacterial pathogens use virulence factors to induce intestinal inflammation, which liberates nutrients for selective use by the infecting microbe. studies of salmonella enterica serovar typhimurium (s. typhimurium) infection in a streptomycin-treated mouse colitis model show how virulence factor-induced inflammation can produce nutrients used ...201222893385
a bioinformatic strategy for the detection, classification and analysis of bacterial autotransporters.autotransporters are secreted proteins that are assembled into the outer membrane of bacterial cells. the passenger domains of autotransporters are crucial for bacterial pathogenesis, with some remaining attached to the bacterial surface while others are released by proteolysis. an enigma remains as to whether autotransporters should be considered a class of secretion system, or simply a class of substrate with peculiar requirements for their secretion. we sought to establish a sensitive search ...201222905239
guillain-barré syndrome associated with pulmonary tuberculosis.a 47-year-old male smoker admitted complaining of progressive bilateral lower-limb and upper-limb weakness, without any numbness. he also complained of productive cough and fever for the last 2 weeks before admission. there were no nasal symptoms, sore throat, night sweats, loss of appetite and weight, nor did he have any gastrointestinal symptoms.201222891003
the apo-structure of the low molecular weight protein-tyrosine phosphatase a (mptpa) from mycobacterium tuberculosis allows for better target-specific drug development.protein-tyrosine phosphatases (ptps) and protein-tyrosine kinases co-regulate cellular processes. in pathogenic bacteria, they are frequently exploited to act as key virulence factors for human diseases. mycobacterium tuberculosis, the causative organism of tuberculosis, secretes a low molecular weight ptp (lmw-ptp), mptpa, which is required for its survival upon infection of host macrophages. although there is otherwise no sequence similarity of lmw-ptps to other classes of ptps, the phosphate ...201222888002
atlast4ss: a curated database for type iv secretion systems.the type iv secretion system (t4ss) can be classified as a large family of macromolecule transporter systems, divided into three recognized sub-families, according to the well-known functions. the major sub-family is the conjugation system, which allows transfer of genetic material, such as a nucleoprotein, via cell contact among bacteria. also, the conjugation system can transfer genetic material from bacteria to eukaryotic cells; such is the case with the t-dna transfer of agrobacterium tumefa ...201222876890
epidemiological association of campylobacter jejuni groups with pathogenicity-associated genetic markers.campylobacter jejuni, the most leading cause for bacterial gastroenteritis worldwide, shows a high genetic diversity among its isolates. recently, we demonstrated the existence of six c. jejuni-groups by combining mlst with six genetic markers. these groups were further characterized by the detection of cj1321-cj1326, fucp, cj0178, cj0755/cfra, ceue, plda, cstii, and cstiii in order (i.) to show further associations between these different genetic markers and mlst ccs. moreover, different studie ...201222873291
biosynthesis of the polymannose lipopolysaccharide o-antigens from escherichia coli serotypes o8 and o9a requires a unique combination of single- and multiple-active site mannosyltransferases.the escherichia coli o9a and o8 o-antigen serotypes represent model systems for the abc transporter-dependent synthesis of bacterial polysaccharides. the o9a and o8 antigens are linear mannose homopolymers containing conserved reducing termini (the primer-adaptor), a serotype-specific repeat unit domain, and a terminator. synthesis of these glycans occurs on the polyisoprenoid lipid-linked primer, undecaprenol pyrophosphoryl-glcpnac, by two conserved mannosyltransferases, wbdc and wbdb, and a se ...201222875852
eukaryotic n-glycosylation occurs via the membrane-anchored c-terminal domain of the stt3p subunit of oligosaccharyltransferase.n-glycosylation is an essential and highly conserved protein modification. in eukaryotes, it is catalyzed by a multisubunit membrane-associated enzyme, oligosaccharyltransferase (ot). we report the high resolution structure of the c-terminal domain of eukaryotic stt3p. unlike its soluble β-sheet-rich prokaryotic counterparts, our model reveals that the c-terminal domain of yeast stt3p is highly helical and has an overall oblate spheroid-shaped structure containing a membrane-embedded region. anc ...201222865878
a comparison of computational methods for identifying virulence factors.bacterial pathogens continue to threaten public health worldwide today. identification of bacterial virulence factors can help to find novel drug/vaccine targets against pathogenicity. it can also help to reveal the mechanisms of the related diseases at the molecular level. with the explosive growth in protein sequences generated in the postgenomic age, it is highly desired to develop computational methods for rapidly and effectively identifying virulence factors according to their sequence info ...201222880014
diversity in the protein n-glycosylation pathways within the campylobacter genus.the foodborne bacterial pathogen, campylobacter jejuni, possesses an n-linked protein glycosylation (pgl) pathway involved in adding conserved heptasaccharides to asparagine-containing motifs of >60 proteins, and releasing the same glycan into its periplasm as free oligosaccharides. in this study, comparative genomics of all 30 fully sequenced campylobacter taxa revealed conserved pgl gene clusters in all but one species. structural, phylogenetic and immunological studies showed that the n-glyco ...201222859570
multifunctional role of dextran sulfate sodium for in vivo modeling of intestinal diseases.inflammatory bowel diseases (ibds) are chronic, relapsing disorders that affect the gastrointestinal tract of millions of people and continue to increase in incidence each year. while several factors have been associated with development of ibds, the exact etiology is unknown. research using animal models of ibds is beginning to provide insights into how the different factors contribute to disease development. oral administration of dextran sulfate sodium (dss) to mice induces a reproducible exp ...201222853702
clinical approach to the diagnostic evaluation of hereditary and acquired neuromuscular diseases.for diagnostic evaluation of a neuromuscular disease, the clinician must be able to obtain a relevant patient and family history and perform focused general, musculoskeletal, neurologic, and functional physical examinations to direct further diagnostic evaluations. laboratory studies for hereditary neuromuscular diseases include the relevant molecular genetic studies. the electromyogram and nerve-conduction studies remain an extension of the physical examination, and help to guide further diagno ...201222938875
luminal hypertonicity and acidity modulate colorectal afferents and induce persistent visceral hypersensitivity.carbohydrate malabsorption such as in lactose intolerance or enteric infection causes symptoms that include abdominal pain. because this digestive disorder increases intracolonic osmolarity and acidity by accumulation of undigested carbohydrates and fermented products, we tested whether these two factors (hypertonicity and acidity) would modulate colorectal afferents in association with colorectal nociception and hypersensitivity. in mouse colorectum-pelvic nerve preparations in vitro, afferent ...201222859365
role of lipid rafts and flagellin in invasion of colonic epithelial cells by shiga-toxigenic escherichia coli o113:h21.shiga-toxigenic escherichia coli (stec) o113:h21 strains that lack the locus of enterocyte effacement (lee) efficiently invade eukaryotic cells in vitro, unlike lee-positive o157:h7 strains. we used a flic deletion mutant of the o113:h21 stec strain 98nk2 (98nk2δflic) to show that invasion of colonic epithelial (hct-8) cells is heavily dependent on production of flagellin, even though adherence to the cells was actually enhanced in the mutant. flagellin binds and signals through toll-like recept ...201222689816
localization of aggregatibacter actinomycetemcomitans cytolethal distending toxin subunits during intoxication of live cells.the cytolethal distending toxin (cdt), produced by some clinically important gram-negative bacterial species, is related to the family of ab-type toxins. three heterologous proteins (cdta, cdtb, and cdtc) and a genotoxin mode of action distinguish the cdt from others in this toxin class. crystal structures of several species-specific cdts have provided a basis for predicting subunit interactions and functions. in addition, empirical studies have yielded significant insights into the in vivo inte ...201222645284
legionella pneumophila pathogenesis in the galleria mellonella infection model.legionella pneumophila is a facultative intracellular human pathogen and the etiological agent of severe pneumonia known as legionnaires' disease. its virulence depends on protein secretion systems, in particular, the dot/icm type iv secretion system (t4ss), which is essential to establish a replication-permissive vacuole in macrophages. the analysis of the role of these systems and their substrates for pathogenesis requires easy-to-use models which approximate human infection. we examined the e ...201222645286
campylobacter jejuni-mediated induction of cc and cxc chemokines and chemokine receptors in human dendritic cells.campylobacter jejuni is a leading worldwide bacterial cause of human diarrheal disease. although the specific molecular mechanisms of c. jejuni pathogenesis have not been characterized in detail, host inflammatory responses are thought to be major contributing factors to the resulting typical acute colitis. the intestinal mucosal chemokine response is particularly important in the initial stages of bacterium-induced gut inflammation. chemokines attract blood phagocytes and lymphocytes to the sit ...201222689814
pediatric renal inflammatory myofibroblastic tumours: a case report and review of the etiology and management options.inflammatory myofibroblastic tumours (imts) have been described in lung, bladder, spleen, breast, pancreas, liver, colon, spermatic cord, prostate, peripheral nerves, orbit and kidney. traditionally believed as having a reactive pathogenesis, imts are now viewed more as a neoplasm. this report describes a case of a renal imt in a 14-year-old girl with spina bifida associated neurogenic bladder and a history of recurrent urinary tract infections. this represents a unique case as pediatric renal i ...201223093568
amixicile, a novel inhibitor of pyruvate: ferredoxin oxidoreductase, shows efficacy against clostridium difficile in a mouse infection model.clostridium difficile infection (cdi) is a serious diarrheal disease that often develops following prior antibiotic usage. one of the major problems with current therapies (oral vancomycin and metronidazole) is the high rate of recurrence. nitazoxanide (ntz), an inhibitor of pyruvate:ferredoxin oxidoreductase (pfor) in anaerobic bacteria, parasites, helicobacter pylori, and campylobacter jejuni, also shows clinical efficacy against cdi. from a library of ∼250 analogues of ntz, we identified lead ...201222585229
automated detection of toxigenic clostridium difficile in clinical samples: isothermal tcdb amplification coupled to array-based detection.clostridium difficile can carry a genetically variable pathogenicity locus (paloc), which encodes clostridial toxins a and b. in hospitals and in the community at large, this organism is increasingly identified as a pathogen. to develop a diagnostic test that combines the strengths of immunoassays (cost) and dna amplification assays (sensitivity/specificity), we targeted a genetically stable paloc region, amplifying tcdb sequences and detecting them by hybridization capture. the assay employs a ...201222675134
fdhtu-modulated formate dehydrogenase expression and electron donor availability enhance recovery of campylobacter jejuni following host cell infection.campylobacter jejuni is a food-borne bacterial pathogen that colonizes the intestinal tract and causes severe gastroenteritis. interaction with host epithelial cells is thought to enhance severity of disease, and the ability of c. jejuni to modulate its metabolism in different in vivo and environmental niches contributes to its success as a pathogen. a c. jejuni operon comprising two genes that we designated fdht (cjj81176_1492) and fdhu (cjj81176_1493) is conserved in many bacterial species. de ...201222636777
selenium-dependent biogenesis of formate dehydrogenase in campylobacter jejuni is controlled by the fdhtu accessory genes.the food-borne bacterial pathogen campylobacter jejuni efficiently utilizes organic acids such as lactate and formate for energy production. formate is rapidly metabolized via the activity of the multisubunit formate dehydrogenase (fdh) enzyme, of which the fdha subunit is predicted to contain a selenocysteine (sec) amino acid. in this study we investigated the function of the cj1500 and cj1501 genes of c. jejuni, demonstrate that they are involved in selenium-controlled production of fdh, and p ...201222609917
avian resistance to campylobacter jejuni colonization is associated with an intestinal immunogene expression signature identified by mrna sequencing.campylobacter jejuni is the most common cause of human bacterial gastroenteritis and is associated with several post-infectious manifestations, including onset of the autoimmune neuropathy guillain-barré syndrome, causing significant morbidity and mortality. poorly-cooked chicken meat is the most frequent source of infection as c. jejuni colonizes the avian intestine in a commensal relationship. however, not all chickens are equally colonized and resistance seems to be genetically determined. we ...201222870198
association of campylobacter jejuni metabolic traits with multilocus sequence types.in this study, we describe the association of three campylobacter jejuni metabolism-related traits, γ-glutamyl-transpeptidase (ggt), fucose permease (fucp), and secreted l-asparaginase [ansb(s)], with multilocus sequence types (sts). a total of 710 c. jejuni isolates with known sts were selected and originated from humans, poultry, bovines, and the environment. among these isolates, we found 31.1% to produce ggt and 49.3% and 30.3% to be positive for ansb(s) and fucp, respectively. the combinati ...201222660710
seasonal diversity of planktonic protists in southwestern alberta rivers over a 1-year period as revealed by terminal restriction fragment length polymorphism and 18s rrna gene library analyses.the temporal dynamics of planktonic protists in river water have received limited attention despite their ecological significance and recent studies linking phagotrophic protists to the persistence of human-pathogenic bacteria. using molecular-based techniques targeting the 18s rrna gene, we studied the seasonal diversity of planktonic protists in southwestern alberta rivers (oldman river basin) over a 1-year period. nonmetric multidimensional scaling analysis of terminal restriction fragment le ...201222685143
arcobacter in lake erie beach waters: an emerging gastrointestinal pathogen linked with human-associated fecal contamination.the genus arcobacter has been associated with human illness and fecal contamination by humans and animals. to better characterize the health risk posed by this emerging waterborne pathogen, we investigated the occurrence of arcobacter spp. in lake erie beach waters. during the summer of 2010, water samples were collected 35 times from the euclid, villa angela, and headlands (east and west) beaches, located along ohio's lake erie coast. after sample concentration, arcobacter was quantified by rea ...201222660704
antimicrobial activities of fidaxomicin.fidaxomicin is bactericidal against clostridium difficile. the combined results of 8 in vitro studies of 1323 c. difficile isolates showed the minimum inhibitory concentration (mic) range of fidaxomicin to be ≤ 0.001-1 μg/ml, with a maximum mic for inhibition of 90% of organisms (mic(90)) of 0.5 μg/ml. isolates from 2 phase iii clinical trials demonstrated that fidaxomicin mics of baseline isolates did not predict clinical cure, failure, or recurrence of c. difficile infections. no resistance to ...201222752863
zinc competition among the intestinal microbiota.bioavailable levels of trace metals, such as iron and zinc, for bacterial growth in nature are sufficiently low that most microbes have evolved high-affinity binding and transport systems. the microbe campylobacter jejuni lives in the gastrointestinal tract of chickens, the principal source of human infection. a high-affinity abc transporter for zinc uptake is required for campylobacter survival in chicken intestines in the presence of a normal microbiota but not when chickens are raised with a ...201222851657
sialoadhesin promotes rapid proinflammatory and type i ifn responses to a sialylated pathogen, campylobacter jejuni.sialoadhesin (sn) is a macrophage (mφ)-restricted receptor that recognizes sialylated ligands on host cells and pathogens. although sn is thought to be important in cellular interactions of mφs with cells of the immune system, the functional consequences of pathogen engagement by sn are unclear. as a model system, we have investigated the role of sn in mφ interactions with heat-killed campylobacter jejuni expressing a gd1a-like, sialylated glycan. compared to sn-expressing bone marrow-derived ma ...201222851711
inhibition of listeria monocytogenes atcc 19115 on ham steak by tea bioactive compounds incorporated into chitosan-coated plastic films.the consumer demands for better quality and safety of food products have given rise to the development and implementation of edible films. the use of antimicrobial films can be a promising tool for controlling l. monocytogenes on ready to eat products. the aim of this study was to develop effective antimicrobial films incorporating bioactive compounds from green and black teas into chitosan, for controlling l. monocytogenes atcc 19115 on vacuum-packaged ham steak. the effectiveness of these anti ...201222839307
guillain-barre syndrome associated with norovirus infection.the authors describe a case of guillain-barre syndrome after the diarrhoeal infection due to norovirus. many infections have been described most notably campylobacter jejuni. to our knowledge a connection to norovirus has not been described in the world literature. the authors speculate on why this may be so.201222736780
interaction effects between sender and receiver processes in indirect transmission of campylobacter jejuni between broilers.infectious diseases in plants, animals and humans are often transmitted indirectly between hosts (or between groups of hosts), i.e. via some route through the environment instead of via direct contacts between these hosts. here we study indirect transmission experimentally, using transmission of campylobacter jejuni (c. jejuni) between spatially separated broilers as a model system. we distinguish three stages in the process of indirect transmission; (1) an infectious "sender" excretes the agent ...201222831274
examination of nanoparticle inactivation of campylobacter jejuni biofilms using infrared and raman spectroscopies.to investigate inactivation effect and mechanism of zinc oxide nanoparticles (zno nps) activity against campylobacter jejuni biofilms.201222734855
genomic and proteomic characterization of sumu, a mu-like bacteriophage infecting haemophilus parasuis.haemophilus parasuis, the causative agent of glässer's disease, is prevalent in swine herds and clinical signs associated with this disease are meningitis, polyserositis, polyarthritis, and bacterial pneumonia. six to eight week old pigs in segregated early weaning herds are particularly susceptible to the disease. insufficient colostral antibody at weaning or the mixing of pigs with heterologous virulent h. parasuis strains from other farm sources in the nursery or grower-finisher stage are con ...201222823751
exploiting the nephrotoxic effects of venom from the sea anemone, phyllodiscus semoni, to create a hemolytic uremic syndrome model in the rat.in the natural world, there are many creatures with venoms that have interesting and varied activities. although the sea anemone, a member of the phylum coelenterata, has venom that it uses to capture and immobilise small fishes and shrimp and for protection from predators, most sea anemones are harmless to man. however, a few species are highly toxic; some have venoms containing neurotoxins, recently suggested as potential immune-modulators for therapeutic application in immune diseases. phyllo ...201222851928
mucosal reactive oxygen species decrease virulence by disrupting campylobacter jejuni phosphotyrosine signaling.reactive oxygen species (ros) play key roles in mucosal defense, yet how they are induced and the consequences for pathogens are unclear. we report that ros generated by epithelial nadph oxidases (nox1/duox2) during campylobacter jejuni infection impair bacterial capsule formation and virulence by altering bacterial signal transduction. upon c. jejuni invasion, ros released from the intestinal mucosa inhibit the bacterial phosphotyrosine network that is regulated by the outer-membrane tyrosine k ...201222817987
diarrhea outbreak during u.s. military training in el salvador.infectious diarrhea remains a major risk to deployed military units worldwide in addition to their impact on travelers and populations living in the developing world. this report describes an outbreak of diarrheal illness in the u.s. military's 130(th) maneuver enhancement brigade deployed in san vicente, el salvador during a training and humanitarian assistance mission. an outbreak investigation team from u.s. naval medical research unit-six conducted an epidemiologic survey and environmental a ...201222815747
planktonic aggregates of staphylococcus aureus protect against common antibiotics.bacterial cells are mostly studied during planktonic growth although in their natural habitats they are often found in communities such as biofilms with dramatically different physiological properties. we have examined another type of community namely cellular aggregates observed in strains of the human pathogen staphylococcus aureus. by laser-diffraction particle-size analysis (lda) we show, for strains forming visible aggregates, that the aggregation starts already in the early exponential gro ...201222815921
viscosity dictates metabolic activity of vibrio ruber.little is known about metabolic activity of bacteria, when viscosity of their environment changes. in this work, bacterial metabolic activity in media with viscosity ranging from 0.8 to 29.4 mpas was studied. viscosities up to 2.4 mpas did not affect metabolic activity of vibrio ruber. on the other hand, at 29.4 mpas respiration rate and total dehydrogenase activity increased 8 and 4-fold, respectively. the activity of glucose-6-phosphate dehydrogenase (gpd) increased up to 13-fold at higher vis ...201222826705
structural heterogeneity of terminal glycans in campylobacter jejuni lipooligosaccharides.lipooligosaccharides of the gastrointestinal pathogen campylobacter jejuni are regarded as a major virulence factor and are implicated in the production of cross-reactive antibodies against host gangliosides, which leads to the development of autoimmune neuropathies such as guillain-barré and fisher syndromes. c. jejuni strains are known to produce diverse los structures encoded by more than 19 types of los biosynthesis clusters. this study demonstrates that the final c. jejuni los structure can ...201222815868
recent advances in small bowel diseases: part i.as is the case in all parts of gastroenterology and hepatology, there have been many advances in our knowledge and understanding of small intestinal diseases. over 1000 publications were reviewed for 2008 and 2009, and the important advances in basic science as well as clinical applications were considered. in part i of this editorial review, seven topics are considered: intestinal development; proliferation and repair; intestinal permeability; microbiotica, infectious diarrhea and probiotics; d ...201222807604
phylogenetic position of aquificales based on the whole genome sequences of six aquificales species.species belonging to the order aquificales are believed to be an early branching lineage within the bacteria. however, the branching order of this group in single-gene phylogenetic trees is highly variable; for example, it has also been proposed that the aquificales should be grouped with ε-proteobacteria. to investigate the phylogenetic position of aquificales at the whole-genome level, here we reconstructed the phylogenetic trees of 18 bacteria including six aquificales species based on the co ...201222844640
glycerol monolaurate antibacterial activity in broth and biofilm cultures.glycerol monolaurate (gml) is an antimicrobial agent that has potent activity against gram-positive bacteria. this study examines gml antibacterial activity in comparison to lauric acid, in broth cultures compared to biofilm cultures, and against a wide range of gram-positive, gram-negative, and non-gram staining bacteria.201222808139
campylobacter jejuni induces acute enterocolitis in gnotobiotic il-10-/- mice via toll-like-receptor-2 and -4 signaling.campylobacter jejuni is a leading cause of foodborne bacterial enterocolitis worldwide. investigation of immunopathology is hampered by a lack of suitable vertebrate models. we have recently shown that gnotobiotic mice as well as conventional il-10(-/-) animals are susceptible to c. jejuni infection and develop intestinal immune responses. however, clinical symptoms of c. jejuni infection were rather subtle and did not reflect acute bloody diarrhea seen in human campylobacteriosis.201222808254
glycopp: a webserver for prediction of n- and o-glycosites in prokaryotic protein sequences.glycosylation is one of the most abundant post-translational modifications (ptms) required for various structure/function modulations of proteins in a living cell. although elucidated recently in prokaryotes, this type of ptm is present across all three domains of life. in prokaryotes, two types of protein glycan linkages are more widespread namely, n- linked, where a glycan moiety is attached to the amide group of asn, and o- linked, where a glycan moiety is attached to the hydroxyl group of se ...201222808107
gene expression analysis of toll-like receptor pathways in heterophils from genetic chicken lines that differ in their susceptibility to salmonella enteritidis.previously conducted studies using two chicken lines (a and b) show that line a birds have increased resistance to a number of bacterial and protozoan challenges and that heterophils isolated from line a birds are functionally more responsive. furthermore, when stimulated with toll-like receptor (tlr) agonists, heterophils from line a expressed a totally different cytokine and chemokine mrna expression pattern than heterophils from line b. a large-scale gene expression profile using an agilent 4 ...201222783275
noncanonical notch signaling modulates cytokine responses of dendritic cells to inflammatory stimuli.dendritic cell (dc)-derived cytokines play a key role in specifying adaptive immune responses tailored to the type of pathogen encountered and the local tissue environment. however, little is known about how dcs perceive the local environment. we investigated whether endogenous notch signaling could affect dc responses to pathogenic stimuli. we demonstrate that concurrent notch and tlr stimulation results in a unique cytokine profile in mouse bone-marrow derived dcs characterized by enhanced il- ...201222753939
modification of the campylobacter jejuni n-linked glycan by eptc protein-mediated addition of phosphoethanolamine.campylobacter jejuni is the major worldwide cause of bacterial gastroenteritis. c. jejuni possesses an extensive repertoire of carbohydrate structures that decorate both protein and non-protein surface-exposed structures. an n-linked glycosylation system encoded by the pgl gene cluster mediates the synthesis of a rigidly conserved heptasaccharide that is attached to protein substrates or released as free oligosaccharide in the periplasm. removal of n-glycosylation results in reduced virulence an ...201222761430
infection as a cause of type 1 diabetes?it is the current opinion that pathogens, such as viruses, are contributing to the development of type 1 diabetes (t1d) in susceptible individuals. this opinion is based on epidemiological associations, direct isolation of pathogens from the islets of langerhans, as well as a large amount of data from various experimental animal models. human enteroviruses have dominated the literature associated with the etiology of t1d. however, virus infections have also been reported to protect from autoimmu ...201222504578
chemoenzymatic synthesis of an isoprenoid phosphate tool for the analysis of complex bacterial oligosaccharide biosynthesis.undecaprenyl pyrophosphate synthase (upps) is a key enzyme that catalyzes the production of bactoprenols, which act as membrane anchors for the assembly of complex bacterial oligosaccharides. one of the major hurdles in understanding the assembly of oligosaccharide assembly is a lack of chemical tools to study this process, since bactoprenols and the resulting isoprenoid-linked oligosaccharides lack handles or chromophores for use in pathway analysis. here we describe the isolation of a new upps ...201222925763
cj1136 is required for lipooligosaccharide biosynthesis, hyperinvasion, and chick colonization by campylobacter jejuni.campylobacter jejuni is a major cause of bacterial food-borne enteritis worldwide, and invasion into intestinal epithelial cells is an important virulence mechanism. recently we reported the identification of hyperinvasive c. jejuni strains and created a number of transposon mutants of one of these strains, some of which exhibited reduced invasion into int-407 and caco-2 cells. in one such mutant the transposon had inserted into a homologue of cj1136, which encodes a putative galactosyltransfera ...201222508861
comparison of medium, temperature, and length of incubation for detection of vancomycin-resistant enterococcus.campylobacter (campy; bd diagnostics, sparks, md), spectra vre (remel, lenexa, ks), and bile-esculin-azide-vancomycin (beav; remel) agars were compared for their ability to detect vancomycin-resistant enterococci (vre) in 750 stool specimens. the media were compared at 24 h and 48 h of incubation at 35°c and 42°c. when incubated for 24 h at 35°c, campy was the most sensitive (97.8%) and specific (99.9%) but was comparable to spectra, which has a sensitivity of 95.6% and a specificity of 99.1%, w ...201222535989
reactive oxygen species in the signaling and adaptation of multicellular microbial communities.one of the universal traits of microorganisms is their ability to form multicellular structures, the cells of which differentiate and communicate via various signaling molecules. reactive oxygen species (ros), and hydrogen peroxide in particular, have recently become well-established signaling molecules in higher eukaryotes, but still little is known about the regulatory functions of ros in microbial structures. here we summarize current knowledge on the possible roles of ros during the developm ...201222829965
complete genome sequence of helicobacter cinaedi strain pagu611, isolated in a case of human bacteremia.we report the complete genome sequence of helicobacter cinaedi strain pagu611, isolated in a case of human bacteremia. the pagu611 genome comprises a 2,078,348-bp chromosome and a 23,054-bp plasmid. the chromosome contains a unique genomic island, encoding a type vi secretion system and clustered regularly interspaced short palindromic repeat (crispr) loci.201222740670
assessment of public health events through international health regulations, united states, 2007-2011.under the current international health regulations, 194 states parties are obligated to report potential public health emergencies of international concern to the world health organization (who) within 72 hours of becoming aware of an event. during july 2007-december 2011, who assessed and posted on a secure web portal 222 events from 105 states parties, including 24 events from the united states. twelve us events involved human influenza caused by a new virus subtype, including the first report ...201222709566
variation of chemosensory receptor content of campylobacter jejuni strains and modulation of receptor gene expression under different in vivo and in vitro growth conditions.chemotaxis is crucial for the colonisation/infection of hosts with campylobacter jejuni. central to chemotaxis are the group a chemotaxis genes that are responsible for sensing the external environment. the distribution of group a chemoreceptor genes, as found in the c. jejuni sequenced strains, tlp1-4, 7, 10 and 11 were determined in 33 clinical human and avian isolates.201222747654
aglr is required for addition of the final mannose residue of the n-linked glycan decorating the haloferax volcanii s-layer glycoprotein.recent studies of haloferax volcanii have begun to elucidate the steps of n-glycosylation in archaea, where this universal post-translational modification remains poorly described. in hfx. volcanii, a series of agl proteins catalyzes the assembly and attachment of a n-linked pentasaccharide to the s-layer glycoprotein. although roles have been assigned to the majority of agl proteins, others await description. in the following, the contribution of aglr to n-glycosylation was addressed.201222750201
ataxia, ophthalmoplegia, and areflexia: what would you think?we present here a case of carcinomatous meningitis presenting as miller fisher syndrome (mfs). there are four further cases described in the literature with evidence of tumour invasion within the central nervous system (cns) shown either in cerebrospinal fluid examination or on histology. there are further five cases described in which an association between cancer and a miller fisher phenotype has been shown. some of these have identified antiganglioside antibodies in the serum and, in one case ...201222934206
Refresh
firstprev
...81828384858687888990...
nextlast
Displaying items 8701 - 8800 of 12694