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chromobacterium violaceum necrotizing fasciitis: a case report and review of the literature.necrotizing fasciitis is a severe, rapidly progressive infection of the subcutaneous tissue that causes significant destruction. it is rarely encountered in the pediatric population. we describe the case of a 14-year-old boy who was diagnosed with chromobacterium violaceum necrotizing fasciitis and subsequently found to have autosomal recessive chronic granulomatous disease.201223288793
doxycycline interferes with quorum sensing-mediated virulence factors and biofilm formation in gram-negative bacteria.inhibition of quorum sensing (qs)-regulated virulence factors including biofilm is a recognized anti-pathogenic drug target. the search for safe and effective anti-qs agents is expected to be useful to combat diseases caused by multidrug-resistant bacteria. in this study, effect of a commonly used antibiotic, doxycycline on qs was evaluated using sensor strains of chromobacterium violaceum (atcc 12472 and cvo26) and pseudomonas aeruginosa pao1. sub-mics of doxycycline reduced qs-controlled viola ...201323299903
dynamic regulation of phenylalanine hydroxylase by simulated redox manipulation.recent clinical studies revealed increased phenylalanine levels and phenylalanine to tyrosine ratios in patients suffering from infection, inflammation and general immune activity. these data implicated down-regulation of activity of phenylalanine hydroxylase by oxidative stress upon in vivo immune activation. though the structural damage of oxidative stress is expected to be comparably small, a structural rationale for this experimental finding was lacking. hence, we investigated the impact of ...201223300845
inflammasomes and host defenses against bacterial infections.the inflammasome has emerged as an important molecular protein complex which initiates proteolytic processing of pro-il-1β and pro-il-18 into mature inflammatory cytokines. in addition, inflammasomes initiate pyroptotic cell death that may be independent of those cytokines. inflammasomes are central to elicit innate immune responses against many pathogens, and are key components in the induction of host defenses following bacterial infection. here, we review recent discoveries related to nlrp1, ...201323318142
ellagic acid derivatives from terminalia chebula retz. downregulate the expression of quorum sensing genes to attenuate pseudomonas aeruginosa pao1 virulence.burgeoning antibiotic resistance in pseudomonas aeruginosa has necessitated the development of anti pathogenic agents that can quench acylhomoserine lactone (ahl) mediated qs with least risk of resistance. this study explores the anti quorum sensing potential of t. chebula retz. and identification of probable compounds(s) showing anti qs activity and the mechanism of attenuation of p. aeruginosa pao1 virulence factors.201323320085
antibiotics at subinhibitory concentrations improve the quorum sensing behavior of chromobacterium violaceum.increasing evidence has shown that antibiotics function as intermicrobial signaling molecules instead of killing weapons. however, mechanisms and key factors that are involved in such functions remain poorly understood. earlier findings have associated antibiotic signaling with quorum sensing (qs); however, results varied among experiments, antibiotics, and bacterial strains. in this study, we found that antibiotics at subinhibitory concentrations improved the violacein-producing ability of chro ...201323330731
crystalline xylitol production by a novel yeast, pichia caribbica (hq222812), and its application for quorum sensing inhibition in gram-negative marker strain chromobacterium violaceum cv026.xylitol, a sugar alcohol, is fast gaining ground over other artificial sugar substitutes owing to its advantageous properties. xylitol is a safer alternative for diabetics because of insulin-independent metabolism. it has beneficial properties suitable to form an important part of odontological formulations. conventional commercial production of xylitol involves harsh chemical method operating at high temperature and pressure. thus, microbial production of xylitol is preferred over chemical meth ...201323338824
quorum sensing inhibition by asparagopsis taxiformis, a marine macro alga: separation of the compound that interrupts bacterial communication.the majority of the marine algal species, though completing their life cycle in seawater, are rarely susceptible to fouling, making them an important source of quorum sensing (qs) inhibitory substances. the separation and characterization of qs inhibitors are crucial for any potential application. thirty marine macroalgae were tested for qs inhibition activity by using chromobacterium violaceum cv026 as the reporter strain, and among them, asparagopsis taxiformis showed antibacterial, as well as ...201323344114
chronic granulomatous disease.chronic granulomatous disease (cgd) is a paradigm for nonlymphoid primary immune defects, and has guided elucidation of oxygen metabolism in the phagocyte, vasculature, and brain. it has been in the forefront of the development of antimicrobial prophylaxis before the advent of advanced hiv and before its routine use in neutropenia. it has been an attractive target for gene therapy and bone marrow transplantation for nonmalignant diseases. therefore, cgd is worthy of attention for its historical ...201223351990
chronic granulomatous disease.chronic granulomatous disease (cgd) is a paradigm for nonlymphoid primary immune defects, and has guided elucidation of oxygen metabolism in the phagocyte, vasculature, and brain. it has been in the forefront of the development of antimicrobial prophylaxis before the advent of advanced hiv and before its routine use in neutropenia. it has been an attractive target for gene therapy and bone marrow transplantation for nonmalignant diseases. therefore, cgd is worthy of attention for its historical ...201223351990
the rise of pathogens: predation as a factor driving the evolution of human pathogens in the environment.bacteria in the environment must survive predation from bacteriophage, heterotrophic protists, and predatory bacteria. this selective pressure has resulted in the evolution of a variety of defense mechanisms, which can also function as virulence factors. here we discuss the potential dual function of some of the mechanisms, which protect against heterotrophic protists, and how predation pressure leads to the evolution of pathogenicity. this is in accordance with the coincidental evolution hypoth ...201323354181
a rare nonfatal presentation of disseminated chromobacterium violaceum sepsis.we present a case of disseminated chromobacterium violaceum sepsis with multiple liver and splenic abscesses presenting with skin lesions and cardiogenic shock, and later diagnosed to have chronic granulomatous disease. the patient was treated with prolonged antimicrobial therapy, after which she recovered and remained asymptomatic on follow-up.201523380618
diabetes, renal and cardiovascular disease in p47 phox-/- chronic granulomatous disease.chronic granulomatous disease is a rare immunodeficiency due to defects in the phagocyte nadph oxidase. the x-linked form (gp91 (phox) deficiency) accounts for about 70 % of cases; autosomal recessive p47 (phox) deficiency accounts for about 25 % of cases. we identified a 10 % incidence of diabetes in p47 (phox) deficient cgd, but none in x-linked cgd. renal and cardiovascular diseases were also higher in p47 (phox) deficiency. p47 (phox) deficient cgd has non-infectious morbidities distinct fro ...201323386289
the janthinobacterium sp. hh01 genome encodes a homologue of the v. cholerae cqsa and l. pneumophila lqsa autoinducer synthases.janthinobacteria commonly form biofilms on eukaryotic hosts and are known to synthesize antibacterial and antifungal compounds. janthinobacterium sp. hh01 was recently isolated from an aquatic environment and its genome sequence was established. the genome consists of a single chromosome and reveals a size of 7.10 mb, being the largest janthinobacterial genome so far known. approximately 80% of the 5,980 coding sequences (cdss) present in the hh01 genome could be assigned putative functions. the ...201323405110
relative contributions of vibrio polysaccharide and quorum sensing to the resistance of vibrio cholerae to predation by heterotrophic protists.protozoan grazing is a major mortality factor faced by bacteria in the environment. vibrio cholerae, the causative agent of the disease cholera, is a natural inhabitant of aquatic ecosystems, and its survival depends on its ability to respond to stresses, such as predation by heterotrophic protists. previous results show that grazing pressure induces biofilm formation and enhances a smooth to rugose morphotypic shift, due to increased expression of vibrio polysaccharide (vps). in addition to neg ...201323441178
anti-quorum sensing potential of crude kigelia africana fruit extracts.the increasing incidence of multidrug-resistant pathogens has stimulated the search for novel anti-virulence compounds. although many phytochemicals show promising antimicrobial activity, their power lies in their anti-virulence properties. thus the quorum sensing (qs) inhibitory activity of four crude kigelia africana fruit extracts was assessed qualitatively and quantitatively using the chromobacterium violaceum and agrobacterium tumefaciens biosensor systems. inhibition of qs-controlled viola ...201323447012
a genomic survey of reb homologs suggests widespread occurrence of r-bodies in proteobacteria.bacteria and eukaryotes are involved in many types of interaction in nature, with important ecological consequences. however, the diversity, occurrence, and mechanisms of these interactions often are not fully known. the obligate bacterial endosymbionts of paramecium provide their hosts with the ability to kill sensitive paramecium strains through the production of r-bodies, highly insoluble coiled protein ribbons. r-bodies have been observed in a number of free-living bacteria, where their func ...201323450193
chromobacterium violaceum: important insights for virulence and biotechnological potential by exoproteomic studies.chromobacterium violaceum is a beta-proteobacterium with high biotechnological potential, found in tropical environments. this bacterium causes opportunistic infections in both humans and animals, that can spread throughout several tissues, quickly leading to the death of the host. genomic studies identified potential mechanisms of pathogenicity but no further studies were done to confirm the expression of these systems. in this study 36 unique protein entries were identified in databank from a ...201323455494
inhibition of quorum sensing in gram-negative bacteria by alkylamine-modified cyclodextrins.n-acylhomoserine lactones (ahls) are used as quorum-sensing (qs) signals by gram-negative bacteria. we have reported that the cyclic oligosaccharides known as cyclodextrins (cds) form inclusion complexes with ahls and disrupt qs signaling. in this study, a series of cd derivatives were designed and synthesized to improve the qs inhibitory activity over that of native cds. the production of the red pigment prodigiosin by serratia marcescens as-1, which is regulated by ahl-mediated qs, was drastic ...201323466297
exploiting quorum sensing to confuse bacterial pathogens.cell-cell communication, or quorum sensing, is a widespread phenomenon in bacteria that is used to coordinate gene expression among local populations. its use by bacterial pathogens to regulate genes that promote invasion, defense, and spread has been particularly well documented. with the ongoing emergence of antibiotic-resistant pathogens, there is a current need for development of alternative therapeutic strategies. an antivirulence approach by which quorum sensing is impeded has caught on as ...201323471618
histone deacetylase inhibitors and pancreatic cancer: are there any promising clinical trials?pancreatic cancer, although not very frequent, has an exceptionally high mortality rate, making it one of the most common causes of cancer mortality in developed countries. pancreatic cancer is difficult to diagnose, allowing few patients to have the necessary treatment at a relatively early stage. despite a marginal benefit in survival, the overall response of pancreatic cancer to current systemic therapy continues to be poor, and new therapies are desperately needed. histone deacetylase (hdac) ...201323482354
endophytic actinomycetes: a novel source of potential acyl homoserine lactone degrading enzymes.several gram-negative pathogenic bacteria employ n-acyl-l-homoserine lactone (hsl) quorum sensing (qs) system to control their virulence traits. degradation of acyl-hsl signal molecules by quorum quenching enzyme (qqe) results in a loss of pathogenicity in qs-dependent organisms. the qqe activity of actinomycetes in rhizospheric soil and inside plant tissue was explored in order to obtain novel strains with high hsl-degrading activity. among 344 rhizospheric and 132 endophytic isolates, 127 (36. ...201323484156
design, synthesis and biological evaluation of n-sulfonyl homoserine lactone derivatives as inhibitors of quorum sensing in chromobacterium violaceum.a novel series of n-sulfonyl homoserine lactone derivatives 5a-l has been designed, synthesized and evaluated for quorum sensing inhibitory activities towards violacein production. of the compounds synthesized, compound 5h was found to possess an excellent level of enantiopurity (99.2% e.e.). the results indicated that compounds bearing an ortho substituent on their phenyl ring exhibited excellent levels of inhibitory activity against violacein production. compounds 5h and 5k in particular, with ...201323486105
production of acylated homoserine lactone by gram-positive bacteria isolated from marine water.acylated homoserine lactone (ahl)-based quorum sensing (qs) has been reported to be present only in gram-negative microorganisms. isolation of a novel gram-positive microorganism from sea water, capable of producing ahl, is reported here. the isolate (genbank: jf915892, designated as mpo) belonging to the exiguobacterium genera is capable of inducing the ahl bioreporters, namely chromobacterium violaceum cv026, agrobacterium tumefaceins a136, and e. coli jm 109(psb1075). this inducer is characte ...201323489290
pharmacological effects of rosa damascena.rosa damascena mill l., known as gole mohammadi in is one of the most important species of rosaceae family flowers. r. damascena is an ornamental plant and beside perfuming effect, several pharmacological properties including anti-hiv, antibacterial, antioxidant, antitussive, hypnotic, antidiabetic, and relaxant effect on tracheal chains have been reported for this plant. this article is a comprehensive review on pharmacological effects of r. damascena. online literature searches were performed ...201123493250
diagnosis and treatment of gastrointestinal disorders in patients with primary immunodeficiency.gastrointestinal disorders such as chronic or acute diarrhea, malabsorption, abdominal pain, and inflammatory bowel diseases can indicate immune deficiency. the gastrointestinal tract is the largest lymphoid organ in the body, so it is not surprising that intestinal diseases are common among immunodeficient patients. gastroenterologists therefore must be able to diagnose and treat patients with primary immunodeficiency. immune-related gastrointestinal diseases can be classified as those that dev ...201323501398
role of the phenylalanine-hydroxylating system in aromatic substance degradation and lipid metabolism in the oleaginous fungus mortierella alpina.mortierella alpina is a filamentous fungus commonly found in soil that is able to produce lipids in the form of triacylglycerols that account for up to 50% of its dry weight. analysis of the m. alpina genome suggests that there is a phenylalanine-hydroxylating system for the catabolism of phenylalanine, which has never been found in fungi before. we characterized the phenylalanine-hydroxylating system in m. alpina to explore its role in phenylalanine metabolism and its relationship to lipid bios ...201323503309
drimendiol, a drimane sesquiterpene with quorum sensing inhibition activity.quorum sensing (qs) is a regulatory mechanism that enables bacteria to make collective decisions such as an increase in virulence factors and biofilm production. inhibitors of qs are important research tools in the discovery of new potential anti-bacterial agents. polygodial, drimenol and drimendiol are drimane sesquiterpenoids isolated from drimys winteri, a chilean native tree. their qs activity, when tested on chromobacterium violaceum atcc 12472, showed that drimendiol is an inhibitor of qs, ...201323513712
reactivation of latent hiv by histone deacetylase inhibitors.latent hiv persists in cd4(+) t cells in infected patients under antiretroviral therapy (art). latency is associated with transcriptional silencing of the integrated provirus and driven, at least in part, by histone deacetylases (hdacs), a family of chromatin-associated proteins that regulate histone acetylation and the accessibility of dna to transcription factors. remarkably, inhibition of hdacs is sufficient to reactivate a fraction of latent hiv in a variety of experimental systems. this bas ...201323517573
piper nigrum, piper betle and gnetum gnemon--natural food sources with anti-quorum sensing properties.various parts of piper nigrum, piper betle and gnetum gnemon are used as food sources by malaysians. the purpose of this study is to examine the anti-quorum sensing (anti-qs) properties of p. nigrum, p. betle and g. gnemon extracts. the hexane, chloroform and methanol extracts of these plants were assessed in bioassays involving pseudomonas aeruginosa pa01, escherichia coli [psb401], e. coli [psb1075] and chromobacterium violaceum cv026. it was found that the extracts of these three plants have ...201323519352
structural studies of pseudomonas and chromobacterium ω-aminotransferases provide insights into their differing substrate specificity.the crystal structures and inhibitor complexes of two industrially important ω-aminotransferase enzymes from pseudomonas aeruginosa and chromobacterium violaceum have been determined in order to understand the differences in their substrate specificity. the two enzymes share 30% sequence identity and use the same amino acceptor, pyruvate; however, the pseudomonas enzyme shows activity towards the amino donor β-alanine, whilst the chromobacterium enzyme does not. both enzymes show activity toward ...201323519665
esce and escg are cochaperones for the type iii needle protein escf of enteropathogenic escherichia coli.type iii secretion systems (t3sss) are central virulence mechanisms used by a variety of gram-negative bacteria to inject effector proteins into host cells. the needle polymer is an essential part of the t3ss that provides the effector proteins a continuous channel into the host cytoplasm. it has been shown for a few t3sss that two chaperones stabilize the needle protein within the bacterial cytosol to prevent its premature polymerization. in this study, we characterized the chaperones of the en ...201323524615
mapk pathway activation leads to bim loss and histone deacetylase inhibitor resistance: rationale to combine romidepsin with an mek inhibitor.to identify molecular determinants of histone deacetylase inhibitor (hdi) resistance, we selected hut78 cutaneous t-cell lymphoma (ctcl) cells with romidepsin in the presence of p-glycoprotein inhibitors to prevent transporter upregulation. resistant sublines were 250- to 385-fold resistant to romidepsin and were resistant to apoptosis induced by apicidin, entinostat, panobinostat, belinostat, and vorinostat. a custom taqman array identified increased insulin receptor (insr) gene expression; imm ...201323532732
selective tryptophan determination using tryptophan oxidases involved in bis-indole antibiotic biosynthesis.a novel tryptophan assay was developed using tryptophan oxidases. although many l-amino acid oxidases (laaos) have been reported to catalyze tryptophan oxidation, most of them have broad substrate specificity and oxidize multiple amino acids besides tryptophan. to obtain a tryptophan-specific laao, we focused on bis-indole antibiotic biosynthesis, a bacterial secondary metabolic pathway. a putative laao from streptomyces sp. tp-a0274, stao involved in staurosporine biosynthesis, was heterologous ...201323545192
antimicrobial resistance and virulence: a successful or deleterious association in the bacterial world?hosts and bacteria have coevolved over millions of years, during which pathogenic bacteria have modified their virulence mechanisms to adapt to host defense systems. although the spread of pathogens has been hindered by the discovery and widespread use of antimicrobial agents, antimicrobial resistance has increased globally. the emergence of resistant bacteria has accelerated in recent years, mainly as a result of increased selective pressure. however, although antimicrobial resistance and bacte ...201323554414
tetra- and penta-acylated lipid a structures of porphyromonas gingivalis lps differentially activate tlr4-mediated nf-κb signal transduction cascade and immuno-inflammatory response in human gingival fibroblasts.porphyromonas gingivalis is a major pathogen of periodontal disease that affects a majority of adults worldwide. increasing evidence shows that periodontal disease is linked to various systemic diseases like diabetes and cardiovascular disease, by contributing to increased systemic levels of inflammation. lipopolysaccharides (lps), as a key virulent attribute of p. gingivalis, possesses significant amount of lipid a heterogeneity containing tetra- (lps1435/1449) and penta-acylated (lps1690) stru ...201323554896
re-annotation of protein-coding genes in 10 complete genomes of neisseriaceae family by combining similarity-based and composition-based methods.in this paper, we performed a comprehensive re-annotation of protein-coding genes by a systematic method combining composition- and similarity-based approaches in 10 complete bacterial genomes of the family neisseriaceae. first, 418 hypothetical genes were predicted as non-coding using the composition-based method and 413 were eliminated from the gene list. both the scatter plot and cluster of orthologous groups (cog) fraction analyses supported the result. second, from 20 to 400 hypothetical pr ...201323571676
hypoxia inducible factor pathway inhibitors as anticancer therapeutics.hypoxia is a significant feature of solid tumor cancers. hypoxia leads to a more malignant phenotype that is resistant to chemotherapy and radiation, is more invasive and has greater metastatic potential. hypoxia activates the hypoxia inducible factor (hif) pathway, which mediates the biological effects of hypoxia in tissues. the hif complex acts as a transcription factor for many genes that increase tumor survival and proliferation. to date, many hif pathway inhibitors indirectly affect hif but ...201323573973
anti-quorum sensing potential of the mangrove rhizophora annamalayana.the present study was carried out to assess the anti-quorum sensing (anti-qs) activity of bark extract obtained from the mangrove plant rhizophora annamalayana kathir. against gram-negative bacteria. in microtitre plate assay, the bark extract at a concentration of 1 mg/ml inhibited the qs-dependent violacein production in chromobacterium violaceum atcc 12472. further, the qs-dependent bioluminescence production in the aquatic bacterial pathogen vibrio harveyi mtcc 3438 was also reduced to the l ...201323591758
caffeine as a potential quorum sensing inhibitor.quorum sensing enables bacteria to control the gene expression in response to the cell density. it regulates a variety of bacterial physiological functions such as biofilm formation, bioluminescence, virulence factors and swarming which has been shown contribute to bacterial pathogenesis. the use of quorum sensing inhibitor would be of particular interest in treating bacterial pathogenicity and infections. in this work, we have tested caffeine as quorum sensing inhibitor by using chromobacterium ...201323598500
characterization of aeromonas hydrophila wound pathotypes by comparative genomic and functional analyses of virulence genes.aeromonas hydrophila has increasingly been implicated as a virulent and antibiotic-resistant etiologic agent in various human diseases. in a previously published case report, we described a subject with a polymicrobial wound infection that included a persistent and aggressive strain of a. hydrophila (e1), as well as a more antibiotic-resistant strain of a. hydrophila (e2). to better understand the differences between pathogenic and environmental strains of a. hydrophila, we conducted comparative ...201323611906
bioactive molecules in soil ecosystems: masters of the underground.complex biological and ecological processes occur in the rhizosphere through ecosystem-level interactions between roots, microorganisms and soil fauna. over the past decade, studies of the rhizosphere have revealed that when roots, microorganisms and soil fauna physically contact one another, bioactive molecular exchanges often mediate these interactions as intercellular signal, which prepare the partners for successful interactions. despite the importance of bioactive molecules in sustainable a ...201323615474
nlrp7 and related inflammasome activating pattern recognition receptors and their function in host defense and disease.host defense requires the maturation and release of the pro-inflammatory cytokines interleukin (il)-1β and il-18 and the induction of pyroptotic cell death, which depends on the activation of inflammatory caspases within inflammasomes by innate immune cells. several cytosolic pattern recognition receptors (prrs) have been implicated in this process in response to infectious and sterile agonists. here we summarize the current knowledge on inflammasome-organizing prrs, emphasizing the recently des ...201323618810
biochemical composition, antimicrobial activities,and anti-quorum-sensing activities of ethanol and ethyl acetate extracts from hypericum connatum lam. (guttiferae).we studied the polyphenol composition and antioxidant activity of the ethanolic and ethyl acetate extracts of hypericum connatum (guttiferae), in addition to their antimicrobial and antiquorum-sensing (aqs) properties. the extracts exhibited a similar polyphenol content and a notable antioxidant activity, which was stronger in the ethanolic extract (ec50=3.2 μg/ml). ultrahigh-performance liquid chromatography analyses revealed different polyphenol profiles, with rutin and apigen being the main c ...201323631492
dialects of the dna uptake sequence in neisseriaceae.in all sexual organisms, adaptations exist that secure the safe reassortment of homologous alleles and prevent the intrusion of potentially hazardous alien dna. some bacteria engage in a simple form of sex known as transformation. in the human pathogen neisseria meningitidis and in related bacterial species, transformation by exogenous dna is regulated by the presence of a specific dna uptake sequence (dus), which is present in thousands of copies in the respective genomes. dus affects transform ...201323637627
assessment of anti-quorum sensing activity for some ornamental and medicinal plants native to egypt.this study investigated the effects of some plant extracts on the bacterial communication system, expressed as quorum sensing (qs) activity. quorum sensing has a directly proportional effect on the amount of certain compounds, such as pigments, produced by the bacteria. alcohol extracts of 23 ornamental and medicinal plants were tested for anti-qs activity by the chromobacterium violaceum assay using the agar cup diffusion method. the screening revealed the anti-qs activity of six plants; namely ...201223641343
quorum-sensing inhibitory compounds from extremophilic microorganisms isolated from a hypersaline cyanobacterial mat.in this study, extremely halophilic and moderately thermophilic microorganisms from a hypersaline microbial mat were screened for their ability to produce antibacterial, antidiatom, antialgal, and quorum-sensing (qs) inhibitory compounds. five bacterial strains belonging to the genera marinobacter and halomonas and one archaeal strain belonging to the genus haloterrigena were isolated from a microbial mat. the strains were able to grow at a maximum salinity of 22-25 % and a maximum temperature o ...201323645384
bacterial calligraphy: a memento for undergraduate research students. 201223653806
identification of bacterial protein o-oligosaccharyltransferases and their glycoprotein substrates.o-glycosylation of proteins in neisseria meningitidis is catalyzed by pgll, which belongs to a protein family including waal o-antigen ligases. we developed two hidden markov models that identify 31 novel candidate pgll homologs in diverse bacterial species, and describe several conserved sequence and structural features. most of these genes are adjacent to possible novel target proteins for glycosylation. we show that in the general glycosylation system of n. meningitidis, efficient glycosylati ...201323658772
state of the art for the biosorption process--a review.in recent years, biosorption process has become an economic and eco-friendly alternative treatment technology in the water and wastewater industry. in this light, a number of biosorbents were developed and are successfully employed for treating various pollutants including metals, dyes, phenols, fluoride, and pharmaceuticals in solutions (aqueous/oil). however, still there are few technical barriers in the biosorption process that impede its commercialization and thus to overcome these problems ...201323666641
antifeedant, larvicidal and growth inhibitory bioactivities of novel polyketide metabolite isolated from streptomyces sp. ap-123 against helicoverpa armigera and spodoptera litura.considerable attention has been paid to actinomycetes, especially the secondary metabolites obtained from streptomyces species, as the best alternatives to chemicals as biological control agents for polyphagous pests such as helicoverpa armigera and spodoptera litura. on the basis of their novel biocontrol attributes, novel polyketide metabolite isolated from marine streptomyces sp. ap-123 exhibited significant antifeedant, larvicidal and growth inhibitory activities against polyphagous pests.201323668716
plant-derived natural products as sources of anti-quorum sensing compounds.quorum sensing is a system of stimuli and responses in relation to bacterial cell population density that regulates gene expression, including virulence determinants. consequently, quorum sensing has been an attractive target for the development of novel anti-infective measures that do not rely on the use of antibiotics. anti-quorum sensing has been a promising strategy to combat bacterial infections as it is unlikely to develop multidrug resistant pathogens since it does not impose any selectio ...201323669710
clinical features and laboratory diagnosis of infection with the potential bioterrorism agents burkholderia mallei and burkholderia pseudomallei.burkholderia mallei and burkholderia pseudomallei are the causative organisms of glanders and melioidosis, respectively. although now rare in western countries, both organisms have recently gained much interest because of their unique potential as bioterrorism agents. these organisms are less familiar to medical and laboratory personnel than other select bioterrorism bacterial agents and thus heightened awareness of glanders and melioidosis is crucial in order to enable adequate emergency prepar ...200723675037
efficacy of aiim, an n-acylhomoserine lactonase, against pseudomonas aeruginosa in a mouse model of acute pneumonia.quorum sensing (qs) in pseudomonas aeruginosa regulates the production of many virulence factors and plays an important role in the pathogenesis of p. aeruginosa infection. n-acyl homoserine lactones (ahl) are major qs signal molecules. recently, a novel ahl-lactonase enzyme, aiim, has been identified. the aim of this study was to evaluate the effect of aiim on the virulence of p. aeruginosa in a mouse model of acute pneumonia. we developed a p. aeruginosa pao1 strain harboring an aiim-expressin ...201323689715
f18ab escherichia coli flagella expression is regulated by acyl-homoserine lactone and contributes to bacterial virulence.to investigate the effect of the quorum sensing (qs)-i system on the expression of virulence factors in shiga toxin producing and verotoxin-producing escherichia coli (stec and vtec), the yeni gene from yersinia enterocolitica was cloned into e. coli f18ab 107/86. recombinant e. coli transformed with yeni produced acyl-homoserine lactone synthase (ahl), as measured using cross-streaking assays with the reporter biosensor strain chromobacterium violaceum cv026. the ai-1 positive recombinant f18ab ...201323693029
macrocyclic drugs and synthetic methodologies toward macrocycles.macrocyclic scaffolds are commonly found in bioactive natural products and pharmaceutical molecules. so far, a large number of macrocyclic natural products have been isolated and synthesized. the construction of macrocycles is generally considered as a crucial and challenging step in the synthesis of macrocyclic natural products. over the last several decades, numerous efforts have been undertaken toward the synthesis of complex naturally occurring macrocycles and great progresses have been made ...201323708234
ecthyma gangrenosum in a neonate: not always pseudomonas.ecthyma gangrenosum (eg) is a cutaneous manifestation of invasive infection usually caused by pseudomonas, but can be caused by many bacteria, fungal and viral infections. we present the first reported case of eg caused by invasive escherichia coli in a neonate. a neonate presented with evidence of sepsis and a rapidly evolving 3×3.5 cm(2) well-circumscribed haemorrhagic and necrotic ulcer on the left groin. there was evidence of decreased perfusion of the lower limb owing to pressure effect of ...201323715836
monoclonal antibody-based immunofluorescence microscopy for the rapid identification of burkholderia pseudomallei in clinical specimens.the diagnosis of melioidosis depends on the culture of burkholderia pseudomallei, which takes at least 48 hours. we used a polyclonal-fitc-based immunofluorescence microscopic assay (pab-ifa) on clinical samples to provide a rapid presumptive diagnosis. this has limitations including photobleaching and batch-to-batch variability. this study evaluated an ifa based on a monoclonal antibody specific to b. pseudomallei (mab-ifa) and alexa fluor 488. a diagnostic evaluation was performed on a prospec ...201323716405
interference of bacterial cell-to-cell communication: a new concept of antimicrobial chemotherapy breaks antibiotic resistance.bacteria use a cell-to-cell communication activity termed "quorum sensing" to coordinate group behaviors in a cell density dependent manner. quorum sensing influences the expression profile of diverse genes, including antibiotic tolerance and virulence determinants, via specific chemical compounds called "autoinducers". during quorum sensing, gram-negative bacteria typically use an acylated homoserine lactone (ahl) called autoinducer 1. since the first discovery of quorum sensing in a marine bac ...201323720655
expression and efficient secretion of a functional chitinase from chromobacterium violaceum in escherichia coli.chromobacterium violaceum is a free-living β-proteobacterium found in tropical and subtropical regions. the genomic sequencing of c. violaceum atcc 12472 has revealed many genes that underpin its adaptability to diverse ecosystems. moreover, c. violaceum genes with potential applications in industry, medicine and agriculture have also been identified, such as those encoding chitinases. however, none of the chitinase genes of the atcc 12472 strain have been subjected to experimental validation. c ...201323725035
presence of acyl-homoserine lactones in 57 members of the vibrionaceae family.the aim of this study was to use a sensitive method to screen and quantify 57 vibrionaceae strains for the production of acyl-homoserine lactones (ahls) and map the resulting ahl profiles onto a host phylogeny.201323725044
inhibition of quorum sensing mediated virulence factors production in urinary pathogen serratia marcescens ps1 by marine sponges.the focal intent of this study was to find out an alternative strategy for the antibiotic usage against bacterial infections. the quorum sensing inhibitory (qsi) activity of marine sponges collected from palk bay, india was evaluated against acyl homoserine lactone (ahl) mediated violacein production in chromobacterium violaceum (atcc 12472), cv026 and virulence gene expressions in clinical isolate serratia marcescens ps1. out of 29 marine sponges tested, the methanol extracts of aphrocallistes ...201223729876
human melioidosis, malawi, 2011.a case of human melioidosis caused by a novel sequence type of burkholderia pseudomallei occurred in a child in malawi, southern africa. a literature review showed that human cases reported from the continent have been increasing.201323735189
n-acylhomoserine lactone-degrading bacteria isolated from hatchery bivalve larval cultures.quorum sensing (qs) systems, which depend on n-acylhomoserine lactone (ahl) signal molecules, mediate the production of virulence factors in many pathogenic microorganisms. one hundred and forty-six bacterial strains, isolated from a bivalve hatchery, were screened for their capacity to degrade five synthetic ahls [n-butyryl-dl-homoserine lactone (c4-hsl), n-hexanoyl-dl-homoserine lactone (c6-hsl), n-octanoyl-dl-homoserine lactone (c8-hsl), n-decanoyl-dl-homoserine lactone (c10-hsl) and n-dodeca ...201323743010
a fatal case of pulmonary chromobacterium violaceum infection in an adult.chromobacterium violaceum is a gram negative, facultative anaerobic coccobacillus. human infections are rare and usually occur after exposure to contaminated soil or water. infections can present with fulminant septicemia, multiple abscesses and rapidly spreading soft tissue infections. here we present a fatal case of pulmonary chromobacterium violaceum infection following aspiration of drain water. treatment with fluroquinolones in combination with either co-trimoxazole or amikacin has been des ...201023756802
structural and functional characterization of mppr, an enduracididine biosynthetic enzyme from streptomyces hygroscopicus: functional diversity in the acetoacetate decarboxylase-like superfamily.the nonproteinogenic amino acid enduracididine is a critical component of the mannopeptimycins, cyclic glycopeptide antibiotics with activity against drug-resistant pathogens, including methicillin-resistant staphylococcus aureus. enduracididine is produced in streptomyces hygroscopicus by three enzymes, mppp, mppq, and mppr. on the basis of primary sequence analysis, mppp and mppq are pyridoxal 5'-phosphate-dependent aminotransferases; mppr shares a low, but significant, level of sequence ident ...201323758195
the pomegranate: effects on bacteria and viruses that influence human health.pomegranates have been known for hundreds of years for their multiple health benefits, including antimicrobial activity. the recent surge in multidrug-resistant bacteria and the possibility of widespread global virus pandemics necessitate the need for additional preventative and therapeutic options to conventional drugs. research indicates that pomegranates and their extracts may serve as natural alternatives due to their potency against a wide range of bacterial and viral pathogens. nearly ever ...201323762148
a metagenomic study highlights phylogenetic proximity of quorum-quenching and xenobiotic-degrading amidases of the as-family.quorum-sensing (qs) signals of the n-acylhomoserine lactone (nahl) class are cleaved by quorum-quenching enzymes, collectively named nahlases. here, functional metagenomics allowed the discovery of a novel bacterial nahlase in a rhizosphere that was treated with γ-caprolactone. as revealed by rrs-dgge and rrs-pyrosequencing, this treatment increased the percentage of the nahl-degrading bacteria and strongly biased the structure of the bacterial community, among which azospirillum dominated. amon ...201323762380
comparison of an automated system with conventional identification and antimicrobial susceptibility testing.the present study was designed to compare a fully automated identification/antibiotic susceptibility testing (ast) system bd phoenix (bd) for its efficacy in rapid and accurate identification and ast with conventional manual methods and to determine if the errors reported in ast, such as the (very major errors) vme (false susceptibility), (major errors) me (false resistance), and (minor errors) mie (intermediate category interpretation) were within the range certified by fda. identification and ...201223762748
integrated nonlinear optical imaging microscope for on-axis crystal detection and centering at a synchrotron beamline.nonlinear optical (nlo) instrumentation has been integrated with synchrotron x-ray diffraction (xrd) for combined single-platform analysis, initially targeting applications for automated crystal centering. second-harmonic-generation microscopy and two-photon-excited ultraviolet fluorescence microscopy were evaluated for crystal detection and assessed by x-ray raster scanning. two optical designs were constructed and characterized; one positioned downstream of the sample and one integrated into t ...201323765294
malyngolide from the cyanobacterium lyngbya majuscula interferes with quorum sensing circuitry.extracts of several cyanobacterial species collected from different marine and estuarine locations predominately in florida (usa), with one sample each from belize and oman, were screened for their ability to disrupt quorum sensing (qs) in the reporter strain chromobacterium violaceum cv017. inhibitory activities were detected in the ethyl acetate : methanol (1:1) extracts of several lyngbya spp., and extracts of lyngbya majuscula contained the strongest qs inhibitory activities. extracts of l. ...201023766278
antibacterial activity of extracellular compounds produced by a pseudomonas strain against methicillin-resistant staphylococcus aureus (mrsa) strains.the emergence of multidrug-resistant bacteria is a world health problem. staphylococcus aureus, including methicillin-resistant s. aureus (mrsa) strains, is one of the most important human pathogens associated with hospital and community-acquired infections. the aim of this work was to evaluate the antibacterial activity of a pseudomonas aeruginosa-derived compound against mrsa strains.201323773484
quorum sensing and phytochemicals.most infectious diseases are caused by bacteria, which proliferate within quorum sensing (qs)-mediated biofilms. efforts to block qs in bacteria and disrupt biofilms have enabled the identification of bioactive molecules that are also produced by plants. this mini review primarily focuses on natural qs inhibitors, which display potential for treating bacterial infections and also enhance the safety of food supply.201323774835
chromobacterium violaceum: a rare bacterium isolated from a wound over the scalp.infection due to chromobacterium violaceum, a large motile gram-negative bacillus, is a rare entity that typically starts with a localized skin infection or localized lymphadenitis after contact with stagnant water or soil. it can progress to fulminating septicemia, with necrotizing metastatic lesions and multiple abscesses in the liver, lung, spleen, skin, lymph nodes, and brain, and result in fatal multiorgan failure. we report a case of a young male with a history of fall from a bike into sta ...201223776815
coral-associated bacteria, quorum sensing disrupters, and the regulation of biofouling.marine biofouling, the settlement of microorganisms and macroorganisms on structures submerged in seawater, although economically detrimental, is a successful strategy for survival in hostile environments, where coordinated bacterial communities establish biofilms via the regulation of quorum sensing (qs) communication systems. the inhibition of qs activity among bacteria isolated from different coral species was investigated to gain further insight into its potency in the attenuation, or even t ...201323777289
anti-biofilm forming and anti-quorum sensing activity of selected essential oils and their main components on food-related micro-organisms.the aim of this study was to investigate the effect of clary sage, juniper, lemon and marjoram essential oils (eos) and their major components on the formation of bacterial and yeast biofilms and on the inhibition of ahl-mediated quorum sensing (qs).201323789847
asymmetric preparation of prim-, sec-, and tert-amines employing selected biocatalysts.this account focuses on the application of ω-transaminases, lyases, and oxidases for the preparation of amines considering mainly work from our own lab. examples are given to access α-chiral primary amines from the corresponding ketones as well as terminal amines from primary alcohols via a two-step biocascade. 2,6-disubstituted piperidines, as examples for secondary amines, are prepared by biocatalytical regioselective asymmetric monoamination of designated diketones followed by spontaneous rin ...201323794796
spatial disaggregation of tick occurrence and ecology at a local scale as a preliminary step for spatial surveillance of tick-borne diseases: general framework and health implications in belgium.the incidence of tick-borne diseases is increasing in europe. sub national information on tick distribution, ecology and vector status is often lacking. however, precise location of infection risk can lead to better targeted prevention measures, surveillance and control.201323800283
matrix-assisted laser desorption ionisation-time of flight mass spectrometry for rapid identification of laribacter hongkongensis.laribacter hongkongensis is a gram-negative, facultative anaerobic, motile, s-shaped, urease-positive bacillus associated with invasive infections in liver cirrhosis patients and community-acquired gastroenteritis. most cases of l hongkongensis infections occur in eastern countries. information is lacking on the usefulness of matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (maldi-tof ms) for the identification of bacteria important in eastern countries. using the bru ...201323814260
the epigenome as a potential mediator of cancer and disease prevention in prenatal development.epigenetic events establish a particular gene expression signature for each cell type during differentiation and fertilization. disruption of these epigenetic programs in response to environmental stimuli during prenatal exposure dysregulates the fetal epigenome, potentially impacting susceptibility to disease later in life (the fetal basis of adult disease). maternal dietary modifications during gestation and lactation play a pivotal role in the period of fetal (re)programming. recently, many s ...201323815143
cis-9-octadecenoic acid from the rhizospheric bacterium stenotrophomonas maltophilia bj01 shows quorum quenching and anti-biofilm activities.quorum quenching (qq) is an effective approach for the prevention of bacterial infections involving biofilms. this study reports the qq and anti-biofilm activities of a rhizospheric bacterium identified as stenotrophomonas maltophilia bj01. the qq activity was demonstrated using chromobacterium violaceum cv026 as a biosensor. a maximum of 95% reduction in violacein production, a quorum sensing-regulated behavior, was observed. gas chromatography-mass spectroscopy of the extract showed that the a ...201323844805
association of bacteria with marine invertebrates: implications for ballast water management.bacteria associated with plankton are of importance in marine bioinvasions and the implementation of ship's ballast water treatment technologies. in this study, epibiotic and endobiotic bacteria associated with zooplankton, including barnacle nauplii, veliger larvae, and adults of the copepod oithona sp., were characterized and quantified. barnacle nauplius and veliger larva harbored ~4.4 × 10(5)cells ind(-1) whereas oithona sp. had 8.8 × 10(5)cells ind(-1). computation of bacterial contribution ...201323846742
quorum quenching in culturable phyllosphere bacteria from tobacco.many gram-negative plant pathogenic bacteria employ a n-acylhomoserine lactone (ahl)-based quorum sensing (qs) system to regulate their virulence traits. a sustainable biocontrol strategy has been developed using quorum quenching (qq) bacteria to interfere with qs and protect plants from pathogens. here, the prevalence and the diversity of qq strains inhabiting tobacco leaf surfaces were explored. a total of 1177 leaf-associated isolates were screened for their ability to disrupt ahl-mediated qs ...201323857057
an additional substrate binding site in a bacterial phenylalanine hydroxylase.phenylalanine hydroxylase (pah) is a non-heme iron enzyme that catalyzes oxidation of phenylalanine to tyrosine, a reaction that must be kept under tight regulatory control. mammalian pah has a regulatory domain in which binding of the substrate leads to allosteric activation of the enzyme. however, the existence of pah regulation in evolutionarily distant organisms, for example some bacteria in which it occurs, has so far been underappreciated. in an attempt to crystallographically characterize ...201323860686
enhancing gold recovery from electronic waste via lixiviant metabolic engineering in chromobacterium violaceum.conventional leaching (extraction) methods for gold recovery from electronic waste involve the use of strong acids and pose considerable threat to the environment. the alternative use of bioleaching microbes for gold recovery is non-pollutive and relies on the secretion of a lixiviant or (bio)chemical such as cyanide for extraction of gold from electronic waste. however, widespread industrial use of bioleaching microbes has been constrained by the limited cyanogenic capabilities of lixiviant-pro ...201323868689
identification in marinomonas mediterranea of a novel quinoprotein with glycine oxidase activity.a novel enzyme with lysine-epsilon oxidase activity was previously described in the marine bacterium marinomonas mediterranea. this enzyme differs from other l-amino acid oxidases in not being a flavoprotein but containing a quinone cofactor. it is encoded by an operon with two genes loda and lodb. the first one codes for the oxidase, while the second one encodes a protein required for the expression of the former. genome sequencing of m. mediterranea has revealed that it contains two additional ...201323873697
identification and characterization of ribn, a novel family of riboflavin transporters from rhizobium leguminosarum and other proteobacteria.rhizobia are symbiotic bacteria able to invade and colonize the roots of legume plants, inducing the formation of nodules, where bacteria reduce atmospheric nitrogen (n2) to ammonia (nh3). riboflavin availability influences the capacity of rhizobia to survive in the rhizosphere and to colonize roots. in this study, we identified the rl1692 gene of rhizobium leguminosarum downstream of a flavin mononucleotide (fmn) riboswitch. rl1692 encodes a putative transmembrane permease with two eama domains ...201323935051
characterization and complete sequence of lactonase enzyme from bacillus weihenstephanensis isolate p65 with potential activity against acyl homoserine lactone signal molecules.acyl homoserine lactones (ahls) are the most common class of quorum sensing signal molecules (autoinducers) that have been reported to be essential for virulence of many relevant pathogenic bacteria such as pseudomonas aeruginosa. new approach for controlling infections of such bacteria is through quorum quenching. in this study, the acyl homoserine lactone inhibitory activity of the crude enzyme from a bacillus weihenstephanensis-isolate p65 was characterized. the crude enzyme was found to have ...201323936779
human naip and mouse naip1 recognize bacterial type iii secretion needle protein for inflammasome activation.inflammasome mediated by central nucleotide-binding and oligomerization domain (nod)-like receptor (nlr) protein is critical for defense against bacterial infection. here we show that type iii secretion system (t3ss) needle proteins from several bacterial pathogens, including salmonella typhimurium, enterohemorrhagic escherichia coli, shigella flexneri, and burkholderia spp., can induce robust inflammasome activation in both human monocyte-derived and mouse bone marrow macrophages. needle protei ...201323940371
screening for antibacterial, antifungal, and anti quorum sensing activity.the plate-hole diffusion assay is an invaluable screening tool to evaluate the antibacterial potential of natural products. it relies on the diffusion of test material from pre-cut wells through agar seeded with bacteria. samples that are capable of inhibiting bacterial growth will produce a clear zone surrounding their well. for the evaluation of antifungal activity of natural products, we describe the broth microdilution method. this assay is performed using a 96 well microtiter tray containin ...201323963914
isolation of extended spectrum β-lactamase (esbl) producing bacteria from urban surface waters in malaysia.this was a preliminary study to test for the presence of multiple antibiotic-resistant extended spectrum β-lactamase (esbl) producing bacteria in malaysian urban surface waters. although the literature review revealed several published papers on clinical esbl isolates in malaysia, none were found on esbl isolates obtained from local surface waters.201323966820
accurate and rapid identification of the burkholderia pseudomallei near-neighbour, burkholderia ubonensis, using real-time pcr.burkholderia ubonensis is an environmental bacterium belonging to the burkholderia cepacia complex (bcc), a group of genetically related organisms that are associated with opportunistic but generally nonfatal infections in healthy individuals. in contrast, the near-neighbour species burkholderia pseudomallei causes melioidosis, a disease that can be fatal in up to 95% of cases if left untreated. b. ubonensis is frequently misidentified as b. pseudomallei from soil samples using selective culturi ...201323967229
systems metabolic engineering of escherichia coli for production of the antitumor drugs violacein and deoxyviolacein.violacein and deoxyviolacein are interesting therapeutics against pathogenic bacteria and viruses as well as tumor cells. in the present work, systems-wide metabolic engineering was applied to target escherichia coli, a widely accepted recombinant host in pharmaceutical biotechnology, for production of these high-value products. the basic producer, e. coli dvio-1, that expressed the vioabce cluster from chromobacterium violaceum under control of the inducible arac system, accumulated 180 mg l(-1 ...201323994489
discovery and activity profiling of thailandepsins a through f, potent histone deacetylase inhibitors, from burkholderia thailandensis e264.three new bicyclic depsipeptides, which are related to the previously reported thailandepsins a (1), b (2) and c (3), were discovered from the culture broth of burkholderia thailandensis e264 when supplemented with amino acid precursors, and were subsequently named as thailandepsins d (4), e (5) and f (6), respectively. enzyme assays showed that 1-6 are potent histone deacetylase (hdac) inhibitors, particularly toward hdac1 which represents class i human hdacs.201223997923
chromobacterium haemolyticum-induced bacteremia in a healthy young man.the genus chromobacterium consists of 7 recognized species. among those, only c. violaceum, commonly found in the soil and water of tropical and subtropical regions, has been shown to cause human infection. although human infection is rare, c. violaceum can cause life-threatening sepsis, with metastatic abscesses, most frequently infecting those who are young and healthy.201324004824
resistance to quorum-quenching compounds.bacteria have the remarkable ability to communicate as a group in what has become known as quorum sensing (qs), and this trait has been associated with important bacterial phenotypes, such as virulence and biofilm formation. bacteria also have an incredible ability to evolve resistance to all known antimicrobials. hence, although inhibition of qs has been hailed as a means to reduce virulence in a manner that is impervious to bacterial resistance mechanisms, this approach is unlikely to be a pan ...201324014536
rapid detection of burkholderia pseudomallei in blood cultures using a monoclonal antibody-based immunofluorescent assay.melioidosis is a severe bacterial infection caused by burkholderia pseudomallei. rapid antimicrobial therapy is necessary to improve patient outcome, which is aided by direct detection of b. pseudomallei in clinical samples. a drawback for all antigen assays is that the number of b. pseudomallei in blood usually falls below the achievable level of detection. we performed a prospective cohort study of 461 patients with 541 blood cultures to evaluate the utility of a pre-incubation step prior to d ...201324019434
2(5h)-furanone: a prospective strategy for biofouling-control in membrane biofilm bacteria by quorum sensing inhibition.biofouling of membranes demands costly periodic cleaning and membrane replacement. a sustainable and environmentally friendly solution for maintenance is not available and would be of great interest for many purposes including economical. as complex biofilm formation by environmental strains is the major cause of biofouling and biofilm formation in most cases are controlled by n-acylhomoserine lactone (ahl)-mediated quorum sensing (qs). an effort was made to understand the appropriateness of 2(5 ...201024031485
rauvolfia grandiflora (apocynaceae) extract interferes with staphylococcal density, enterotoxin production and antimicrobial activity.staphylococci bacteria are involved in many human and animal infections and development of alternative antimicrobial drugs against pathogenic bacteria is of great interest to the pharmaceutical industry. this study investigated the in vitro effect of rauvolfia grandiflora methanol extract (root bark fraction) (rge) on the density of atcc strains of staphylococcus aureus and staphylococcus epidermidis, and a clinical enterotoxin-producer, s. aureus bovine strain. the alkaloid, isoreserpiline, obt ...201024031536
enterobacteriaceae in mouth and cloaca of podocnemis expansa and p. unifilis (testudines: chelonia) populations of national park of araguaia plains, brazil.shigella flexnerii and escherichia coli were the most frequent gram-negative bacteria found in the mouth cavity and cloacae of the turtles podocnemis expansa and p. unifilis on beaches in the national park of araguaia, brazil. reptiles are known as salmonella carriers, despite rarely isolated in these turtles.201124031664
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