Publications

surveillance for foodborne disease outbreaks --- united states, 2007.	foodborne agents cause an estimated 76 million illnesses annually in the united states. outbreak surveillance provides insights into the causes of foodborne illness, types of implicated foods, and settings of foodborne infections that can be used in food safety strategies to prevent and control foodborne disease. cdc collects data on foodborne disease outbreaks submitted from all states and territories. this report summarizes epidemiologic data for the 1,097 reported outbreaks occurring during 2 ...	2010	20703204
development and application of a new method for specific and sensitive enumeration of spores of nonproteolytic clostridium botulinum types b, e, and f in foods and food materials.	the highly potent botulinum neurotoxins are responsible for botulism, a severe neuroparalytic disease. strains of nonproteolytic clostridium botulinum form neurotoxins of types b, e, and f and are the main hazard associated with minimally heated refrigerated foods. recent developments in quantitative microbiological risk assessment (qmra) and food safety objectives (fso) have made food safety more quantitative and include, as inputs, probability distributions for the contamination of food materi ...	2010	20709854
prevalence of clostridium perfringens in faeces and ileal contents from grass sickness affected horses: comparisons with 3 control populations.	while previous studies have demonstrated an association between equine grass sickness (egs) and the presence of clostridium botulinum within ileal contents and faeces, no such associations with other intestinal-derived anaerobic bacteria have been extensively investigated.	2010	20716188
the osmolyte trimethylamine n-oxide (tmao) increases the proteolytic activity of botulinum neurotoxin light chains a, b, and e: implications for enhancing analytical assay sensitivity.	botulism, the disease caused by botulinum neurotoxins (bonts), secreted by the spore-forming, anaerobic bacteria clostridium botulinum, has been associated with food poisoning for centuries. in addition, the potency of bonts coupled with the current political climate has produced a threat of intentional, malicious poisoning by these toxins. the ability to detect and measure bonts in complex matrixes is among the highest research priorities. however, the extreme potency of these toxins necessitat ...	2010	20720092
[infant botulism in france, 1991-2009].	infant botulism is caused by the ingestion of spores of clostridium botulinum and affects newborns and infants under 12 months of age. ingested spores multiply and produce botulinum toxin in the digestive tract, which then induces clinical symptoms. a single french case was described in the literature prior to 1991. we describe the cases of infant botulism identified in france between 1991 and 2009. all clinical suspicions of botulism must be declared in france. biological confirmation of the di ...	2010	20724121
coherent pipeline for biomarker discovery using mass spectrometry and bioinformatics.	robust biomarkers are needed to improve microbial identification and diagnostics. proteomics methods based on mass spectrometry can be used for the discovery of novel biomarkers through their high sensitivity and specificity. however, there has been a lack of a coherent pipeline connecting biomarker discovery with established approaches for evaluation and validation. we propose such a pipeline that uses in silico methods for refined biomarker discovery and confirmation.	2010	20796299
rare case of dysphagia.	wound botulism is exceedingly rare and occurs almost exclusively among injection drug users. in 2008 there was a case of wound botulism in a noninjecting drug user reported to the centers for disease control and prevention (cdc). we report a case of a caucasian male developing dysphagia due to wound botulism after having a motorcycle accident that left him with open fractures. the cdc was contacted and the patient was transferred to medical intensive care unit to be emergently started on hepatav ...	2010	20809173
a bacteriophage-based platform for rapid trace detection of proteases.	sensitive, inexpensive, and rapid protease activity assays are of great merit for clinical diagnostics. detection of protease-based toxins produced by clostridium botulinum and bacillus anthracis represents a particularly challenging task, as exceptional sensitivity is a prerequisite because of the extreme potency of the toxins. here we present an inexpensive and sensitive assay platform for activity-based protease quantification utilizing filamentous bacteriophage as an exponentially amplifiabl ...	2010	20812737
targeting botulinum neurotoxin persistence by the ubiquitin-proteasome system.	botulinum neurotoxins (bonts) are the most potent natural toxins known. the effects of bont serotype a (bont/a) can last several months, whereas the effects of bont serotype e (bont/e), which shares the same synaptic target, synaptosomal-associated protein 25 (snap25), last only several weeks. the long-lasting effects or persistence of bont/a, although desirable for therapeutic applications, presents a challenge for medical treatment of bont intoxication. although the mechanisms for bont toxicit ...	2010	20823219
relationships between local carcass density and risk of mortality in molting mallards during avian botulism outbreaks.	removal of bird carcasses has been advocated for management of clostridium botulinum outbreaks on lakes in north america because a reduction in density of toxin-laden maggots produced within bird carcasses is assumed to enhance survival of healthy birds. this inverse relationship between carcass density and survival has been reported in controlled studies with mallards (anas platyrhynchos) but has not been verified in wild ducks during naturally occurring botulism outbreaks. therefore, we radio- ...	2010	20688643
survival of radio-marked mallards in relation to management of avian botulism.	avian botulism outbreaks are frequently perpetuated by type c toxin produced by clostridium botulinum proliferating in decomposing bird carcasses and consumption of toxic maggots from these carcasses by healthy birds. therefore, removing bird carcasses has been advocated for disease management because availability of toxic maggots should be reduced, increasing duck survival. however, this management is expensive, and its effect on waterfowl mortality under field conditions is unknown. we radio-m ...	2010	20688692
an outbreak of type c botulism in waterbirds: incheon, korea.	five outbreaks of botulism in waterbirds were encountered over a 5-yr period from 2004 to 2008 in korea. in october 2008, an outbreak of avian type c botulism affected approximately 2,000 wild waterbirds in the namdong flood control basin, incheon, south korea. ecologic conditions, clinical signs exhibited by moribund birds, and lack of gross pathology and microbial evidence of infectious disease, suggested botulinum intoxication. type c botulinum toxin was demonstrated in duck sera, liquid cult ...	2010	20688696
outbreak of botulism (clostridium botulinum type c) in wild waterfowl: seoul, korea.	over a 6-day period beginning on 15 october 2008, 93 dead or sick wild waterfowl, including mallards, spotbills, and teal species, were found along the shore of a branch stream of the hangang river, which flows through seoul, korea, and were submitted to the national veterinary research and quarantine service (nvrqs) for diagnosis. clinically, the affected birds showed flaccid paralysis of the legs and wings and paralysis of the neck. grossly, no bird showed any lesions, but all had almost empty ...	2010	20688704
clostridium botulinum c2 toxin is internalized by clathrin- and rho-dependent mechanisms.	clostridium botulinum c2 toxin is an adp-ribosyltransferase, causing depolymerization of the actin cytoskeleton in eukaryotic cells. the c2 toxin is a binary toxin consisting of the enzymatic subunit c2i and the binding subunit c2ii. proteolytical activation of the binding subunit triggers the formation of heptameric structures (c2iia), which bind to cellular receptors. c2i is able to bind to c2iia oligomers, and it has been suggested that the whole complex is internalized by a raft-dependent me ...	2010	20690924
inhibition of rho-rock signaling induces apoptotic and non-apoptotic ps exposure in cardiomyocytes via inhibition of flippase.	subsequent to myocardial infarction, cardiomyocytes within the infarcted areas and border zones expose phosphatidylserine (ps) in the outer plasma membrane leaflet (flip-flop). we showed earlier that in addition to apoptosis, this flip-flop can be reversible in cardiomyocytes. we now investigated a possible role for rho and downstream effector rho-associated kinase (rock) in the process of (reversible) ps exposure and apoptosis in cardiomyocytes. in rat cardiomyoblasts (h9c2 cells) and isolated ...	2010	20691698
endopeptidase activities of botulinum neurotoxin type b complex, holotoxin, and light chain.	botulinum neurotoxin (bont) serotype b (bont/b) is one of the serotypes of bont that causes deadly human botulism, though it is used clinically for treatment of many neuromuscular diseases. bont/b is produced by clostridium botulinum, and it is secreted along with a group of neurotoxin-associated proteins (naps) in the form of a bont/b complex. the complex dissociates into a 150-kda holotoxin and naps at alkaline phs. the 150-kda bont/b holotoxin can be nicked to produce a 50-kda domain referred ...	2010	20693440
content of botulinum neurotoxin in botox(r)/vistabel(r), dysport(r)/azzalure(r), and xeomin(r)/bocouture(r).	background: botulinum neurotoxin type a (bont/a) is the active substance in preparations used for the highly effective treatment of neurologic disorders such as cervical dystonia, blepharospasm, or spasticity, as well as other indications such as axillary and palmar hyperhidrosis, and urologic disorders. objective: to determine the amount of bont/a protein present in pharmaceutical preparations of botox(r), dysport(r), and xeomin(r), which are identical with vistabel(r), azzalure(r), and bocoutu ...	2010	20698714
molecular mechanisms of substrate recognition and specificity of botulinum neurotoxin serotype f.	bonts (botulinum neurotoxins) are both deadly neurotoxins and natural toxins that are widely used in protein therapies to treat numerous neurological disorders of dystonia and spinal spasticity. understanding the mechanism of action and substrate specificity of bonts is a prerequisite to develop antitoxin and novel bont-derived protein therapy. to date, there is a lack of detailed information with regard to how bonts recognize and hydrolyse the substrate vamp-2 (vesicle-associated membrane prote ...	2010	21029044
clostridium botulinum neurotoxin type b is heat-stable in milk and not inactivated by pasteurization.	foodborne botulism is caused by the ingestion of foods containing botulinum neurotoxins (bonts). to study the heat stability of clostridium botulinum neurotoxins, we needed to measure and compare the activity of botulinum neurotoxins, serotypes a and b, under various pasteurization conditions. currently, the only accepted assay to detect active c. botulinum neurotoxin is an in vivo mouse bioassay, which raises ethical concerns with regard to the use of experimental animals. in this study, noninv ...	2010	21053906
the c2-streptavidin delivery system promotes the uptake of biotinylated molecules in macrophages and t-leukemia cells.	macrophages are tightly associated with inflammatory diseases as well as carcinogenesis, and therefore represent promising targets for drug delivery and gene transfer. we have recently established a novel protein delivery system based on the binary c2 toxin of clostridium botulinum and streptavidin, allowing the uptake of exogenous biotinylated molecules into mammalian cells. here, we applied this c2-streptavidin delivery system to macrophages and other leukocytes. first, the effect of wild-type ...	2010	20868225
light chain separated from the rest of the type a botulinum neurotoxin molecule is the most catalytically active form.	botulinum neurotoxins (bont) are the most potent of all toxins. the 50 kda n-terminal endopeptidase catalytic light chain (lc) of bont is located next to its central, putative translocation domain. after binding to the peripheral neurons, the central domain of bont helps the lc translocate into cytosol where its proteolytic action on snare (soluble nsf attachment protein receptor) proteins blocks exocytosis of acetyl choline leading to muscle paralysis and eventual death. the translocation domai ...	2010	20877571
in vivo molecular imaging analysis of a nasal vaccine that induces protective immunity against botulism in nonhuman primates.	nasal administration is an effective route for a needle-free vaccine. however, nasally administered ags have the potential to reach the cns directly from the nasal cavity, thus raising safety concerns. in this study, we performed real-time quantitative tracking of a nasal vaccine candidate for botulism, which is a nontoxic subunit fragment of clostridium botulinum type a neurotoxin (bohc/a) effective in the induction of the toxin-neutralizing immune response, by using (18)f-labeled bohc/a-positr ...	2010	20881188
detection and confirmation of clostridium botulinum in water used for cooling at a plant producing low-acid canned foods.	our laboratory tested water samples used for cooling low-acid canned foods at a canning facility under investigation by the u.s. food and drug administration. we used an enzyme-linked immunosorbent assay with digoxigenin-labeled antibodies (dig-elisa) and real-time pcr as screening methods and confirmed the presence of neurotoxin-producing clostridium botulinum in the samples by mouse bioassay.	2010	20889791
purification of a recombinant heavy chain fragment c vaccine candidate against botulinum serotype c neurotoxin [rbontc(h(c))] expressed in pichia pastoris.	a purification process for the manufacture of a recombinant c-terminus heavy chain fragment from botulinum neurotoxin serotype c [rbontc(h(c))], a potential vaccine candidate, has been defined and successfully scaled-up. the rbontc(h(c)) was produced intracellularly in pichia pastoris x-33 using a three step fermentation process, i.e., glycerol batch phase, a glycerol fed-batch phase to achieve high cell densities, followed by a methanol induction phase. the rbontc(h(c)) was captured from the so ...	2010	20933089
universal and specific quantitative detection of botulinum neurotoxin genes.	clostridium botulinum, an obligate anaerobic spore-forming bacterium, produces seven antigenic variants of botulinum toxin that are distinguished serologically and termed "serotypes". botulinum toxin blocks the release of acetylcholine at neuromuscular junctions resulting in flaccid paralysis. the potential lethality of the disease warrants a fast and accurate means of diagnosing suspected instances of food contamination or human intoxication. currently, the food and drug administration (fda)-ac ...	2010	20961439
production and characterisation of a neutralising chimeric antibody against botulinum neurotoxin a.	botulinum neurotoxins, produced by clostridium botulinum bacteria, are the causative agent of botulism. this disease only affects a few hundred people each year, thus ranking it among the orphan diseases. however, botulinum toxin type a (bont/a) is the most potent toxin known to man. due to their potency and ease of production, these toxins were classified by the centers for disease control and prevention (cdc) as category a biothreat agents. for several biothreat agents, like bont/a, passive im ...	2010	20967241
endocytosis of microperoxidase in marginal cells is mainly regulated by rhoa signaling cascade, but not by rho-associated protein kinase, myosin light-chain kinase and myosin phosphatase.	endocytosis plays an important role in cell function and the activation and propagation of signaling pathways. signaling occurs on endocytic pathways and signaling endosomes, and endocytosis is subjected to high-order regulation by cellular signaling mechanisms. marginal cells showed active endocytosis of microperoxidase (mpo) via the clathrin-independent pathway. we examined the signaling pathway that regulates mpo endocytosis in marginal cells using specific inhibitors and activators of signal ...	2010	20975313
rhoa/rho kinase pathway contributes to the pathogenesis of thermal hyperalgesia in diabetic mice.	diabetic neuropathy is one of the most common complications of diabetes and causes various problems in daily life. several investigations have noted that many factors in the spinal cord are involved in the symptoms of painful diabetic neuropathy, and there are very few effective therapeutic regimens. in the present study, we sought to elucidate the role of the rhoa/rho kinase (rock) pathway in thermal hyperalgesia in diabetic mice. the intracellular localization of rhoa and the expression of eno ...	2010	20980102
science, practice, and human errors in controlling clostridium botulinum in heat-preserved food in hermetic containers.	the incidence of botulism in canned food in the last century is reviewed along with the background science; a few conclusions are reached based on analysis of published data. there are two primary aspects to botulism control: the design of an adequate process and the delivery of the adequate process to containers of food. the probability that the designed process will not be adequate to control clostridium botulinum is very small, probably less than 1.0 x 10(-6), based on containers of food, whe ...	2010	20501056
effect of media, additives, and incubation conditions on the recovery of high pressure and heat-injured clostridium botulinum spores.	the effect of additives and post-treatment incubation conditions on the recovery of high pressure and heat-injured (i.e., processed at 620 mpa and 95 and 100 degrees c for 5 min) spores of clostridium botulinum strains, 62-a (proteolytic type a) and 17-b (nonproteolytic type b) was studied. high pressure and heat-injured spores were inoculated into tpgy (trypticase-peptone-glucose-yeast extract) anaerobic broth media containing additives (lysozyme, l-alanine, l-aspartic acid, dipicolonic acid, s ...	2010	20510779
sequence diversity of genes encoding botulinum neurotoxin type f.	botulism due to type f botulinum neurotoxin (bont/f) is rare (<1% of cases), and only a limited number of clostridial strains producing this toxin type have been isolated. as a result, analysis of the diversity of genes encoding bont/f has been challenging. in this study, the entire bont/f nucleotide sequences were determined from 33 type f botulinum toxin-producing clostridial strains isolated from environmental sources and botulism outbreak investigations. we examined proteolytic and nonproteo ...	2010	20511432
thioamides in nature: in search of secondary metabolites in anaerobic microorganisms.		2010	20512793
expression of hc fragment from clostridium botulinum neurotoxin serotype b in escherichia coli and its use as a good immunogen.	a good immunogen was developed as an effective recombinant vaccine candidate that protected mice against botulinum neurotoxin serotype b (bont/b) intoxication. the hc fragment of clostridium botulinum neurotoxin type b (rbont/b-hc) was cloned into the bacterial expression vector pqe-30, and the resulting vector was successfully expressed in the escherichia coli m15 strain. the purified rbont/b-hc protein was used to vaccinate mice and evaluate their survival against challenge with native bont/b. ...	2010	20519939
a novel multiplex pcr method for clostridium botulinum neurotoxin type a gene cluster typing.	a rapid, simple and sensitive multiplex pcr method for bont/a gene cluster typing was developed by combining the results of bont/a subtype (bont/a1 or /a2) gene detection with ha33 and/or p47 gene detection. ten isolates associated with infant botulism in japan were examined and divided into bont/a gene cluster types 2 and 3 by origin (honey feeding or not) and period (1986-1987 or 1999-2007). it is suggested that this multiplex pcr method will be be useful for epidemiological studies of botulis ...	2010	20536728
real-time pcr for clostridium botulinum type c neurotoxin (bontc) gene, also covering a chimeric c/d sequence--application on outbreaks of botulism in poultry.	in recent years, botulism type c has become a serious problem in poultry flocks in sweden. a real-time pcr assay for clostridium botulinum (c. botulinum) type c neurotoxin (bontc) gene was developed as an alternative to the mouse bioassay for detection and identification of c. botulinum type c. the complete method consists of an optimized enrichment protocol followed by automated dna extraction prior to real-time pcr. the sensitivity of the pcr assay was determined with purified dna to approxima ...	2010	20537470
myasthenia and related disorders of the neuromuscular junction.	our understanding of transmission at the neuromuscular junction has increased greatly in recent years. we now recognise a wide variety of autoimmune and genetic diseases that affect this specialised synapse, causing muscle weakness and fatigue. these disorders greatly affect quality of life and rarely can be fatal. myasthenia gravis is the most common disorder and is most commonly caused by autoantibodies targeting postsynaptic acetylcholine receptors. antibodies to muscle-specific kinase (musk) ...	2010	20547629
detection of botulinum neurotoxin serotype b at sub mouse ld(50) levels by a sandwich immunoassay and its application to toxin detection in milk.	botulinum neurotoxin (bont), the causative agent of botulism, a serious neuroparylatic disease, is produced by the anaerobic bacterium clostridium botulinum and consists of a family of seven serotypes (a-h). we previously reported production of high-affinity monoclonal antibodies to bont serotype a.	2010	20548779
conjugative botulinum neurotoxin-encoding plasmids in clostridium botulinum.	clostridium botulinum produces seven distinct serotypes of botulinum neurotoxins (bonts). the genes encoding different subtype neurotoxins of serotypes a, b, f and several dual neurotoxin-producing strains have been shown to reside on plasmids, suggesting that intra- and interspecies transfer of bont-encoding plasmids may occur. the objective of the present study was to determine whether these c. botulinum bont-encoding plasmids are conjugative.	2010	20552020
wound botulism in drug users: a still underestimated diagnosis.	wound botulism is a rare infectious disease that is becoming a frequent complication of parental drug use. diagnosis is often difficult and based on clinical suspicion. we report the first italian case of wound botulism due to intramuscular heroin injection in a 48-year-old man with an acute onset of slurred speech and dysphagia. the most considerable finding of electrophysiological study was the reduction in amplitude of compound muscle action potential which should be considered a useful initi ...	2010	20552239
nanogel antigenic protein-delivery system for adjuvant-free intranasal vaccines.	nanotechnology is an innovative method of freely controlling nanometre-sized materials. recent outbreaks of mucosal infectious diseases have increased the demands for development of mucosal vaccines because they induce both systemic and mucosal antigen-specific immune responses. here we developed an intranasal vaccine-delivery system with a nanometre-sized hydrogel ('nanogel') consisting of a cationic type of cholesteryl-group-bearing pullulan (cchp). a non-toxic subunit fragment of clostridium ...	2010	20562880
multilocus outbreak of foodborne botulism linked to contaminated sausage in hebei province, china.	in 2007, an outbreak of foodborne botulism occurred in hebei province, china. an epidemiological investigation and laboratory detection studies showed that sausage contaminated by type a clostridium botulinum caused this outbreak of food poisoning. its clinical and epidemiological features were different from previous reports of food poisoning.	2010	20569065
[infant botulism].	infant botulism is a rare disease that affects infants below the age of 12 months following absorption of neurotoxins produced by ingested clostridium botulinum spores. the clinical manifestations are caused by symmetrical cranial nerve palsies followed by descending, symmetric flaccid paralysis of voluntary muscles. presenting symptoms include constipation, lethargy, mydriasis and ptosis. the diagnosis is made on the basis of clinical examination and confirmed by isolating the toxin in serum or ...	2010	20569659
clostridiolysin s, a post-translationally modified biotoxin from clostridium botulinum.	through elaboration of its botulinum toxins, clostridium botulinum produces clinical syndromes of infant botulism, wound botulism, and other invasive infections. using comparative genomic analysis, an orphan nine-gene cluster was identified in c. botulinum and the related foodborne pathogen clostridium sporogenes that resembled the biosynthetic machinery for streptolysin s, a key virulence factor from group a streptococcus responsible for its hallmark beta-hemolytic phenotype. genetic complement ...	2010	20581111
single-breath-count test: an important adjunct in the triaging of patients in a mass-casualty incident due to botulism.	clostridium botulinum toxins, the most poisonous substance known to humankind, are considered to be a [us] centers for disease control and prevention category a bioterrorist agent. despite this concern, little has been published with regard to the tactical aspects of triaging a mass-casualty event involving botulism victims arriving at an emergency department. because neuromuscular-ventilatory failure is a principal reason for botulism's early morbidity and mortality, using a quick and sensitive ...	2010	20586014
clostron-targeted mutagenesis.	members of the genus clostridium have long been recognised as important to humankind and its animals, both in terms of the diseases they cause and the useful biological processes they undertake. this has led to increasing efforts directed at deriving greater information on their basic biology, most notably through genome sequence. accordingly, annotated sequences of all of the most important species are now available. however, full exploitation of the data generated has been hindered by the lack ...	2010	20597009
diversity of spore-forming bacteria in cattle manure, slaughterhouse waste and samples from biogas plants.	as biowaste intended for biogas production can contain pathogenic micro-organisms, the recommended treatment is pasteurization at 70°c for 60min. this reduces pathogens such as salmonella spp., whereas spore-forming bacteria (bacillus spp. and clostridium spp.) survive. most spore-forming bacteria are harmless, but some can cause diseases such as blackleg, botulism and anthrax. in this study, the effect of the biogas process on bacillus spp. and clostridium spp. was investigated.	2010	20629803
occurrence and role of lactic acid bacteria in seafood products.	lactic acid bacteria (lab) in fish flesh has long been disregarded because the high post-mortem ph, the low percentage of sugars, the high content of low molecular weight nitrogenous molecules and the low temperature of temperate waters favor the rapid growth of ph-sensitive psychrotolerant marine gram-negative bacteria like pseudomonas, shewanella and photobacterium. in seafood packed in both vacuum (vp) and modified atmosphere (map) packaging commonly co(2) enriched, the growth of the gram-neg ...	2010	20630312
camelid single domain antibodies (vhhs) as neuronal cell intrabody binding agents and inhibitors of clostridium botulinum neurotoxin (bont) proteases.	botulinum neurotoxins (bonts) function by delivering a protease to neuronal cells that cleave snare proteins and inactivate neurotransmitter exocytosis. small (14 kda) binding domains specific for the protease of bont serotypes a or b were selected from libraries of heavy chain only antibody domains (vhhs or nanobodies) cloned from immunized alpacas. several vhhs bind the bont proteases with high affinity (k(d) near 1 nm) and include potent inhibitors of bont/a protease activity (k(i) near 1 nm) ...	2010	20637220
efficacy of a potential trivalent vaccine based on hc fragments of botulinum toxins a, b, and e produced in a cell-free expression system.	botulinum toxins produced by the anaerobic bacterium clostridium botulinum are the most potent biological toxins in nature. traditionally, people at risk are immunized with a formaldehyde-inactivated toxin complex. second generation vaccines are based on the recombinant carboxy-terminal heavy-chain (hc) fragment of the neurotoxin. however, the materialization of this approach is challenging, mainly due to the high at content of clostridial genes. herein, we present an alternative strategy in whi ...	2010	20357058
practical applications of a new botulinum toxin.	the injection of clostridium botulinum type a neurotoxins is among the most commonly performed cosmetic procedures, both in the u.s. and worldwide. the u.s. food and drug administration (fda) approval of a new botulinum neurotoxin type a in april 2009 (bont-a, dysport, medicis, scottsdale, az-hereafter referred to as "dysport") has broadened the neurotoxin market and provides new therapeutic alternatives to practitioners. the introduction of this product raises questions about how to best use it ...	2010	20361474
the case of botulinum toxin in milk: experimental data.	botulinum neurotoxin (bont) is the most toxic substance known to man and the causative agent of botulism. due to its high toxicity and the availability of the producing organism clostridium botulinum, bont is regarded as a potential biological warfare agent. because of the mild pasteurization process, as well as rapid product distribution and consumption, the milk supply chain has long been considered a potential target of a bioterrorist attack. since, to our knowledge, no empirical data on the ...	2010	20363798
botulinum toxin: examining duration of effect in facial aesthetic applications.	patient satisfaction with botulinum toxin treatment is a key success factor in aesthetic procedures and is governed by the interaction of numerous variables. duration of effect is important because it influences retreatment intervals as well as affecting cost and convenience to the patient. in order to review the evidence on the duration of benefit associated with various commercial formulations of botulinum toxin, medline was searched using the following terms: 'botulinum' and 'duration'/'retre ...	2010	20369902
a quantitative real-time pcr method for monitoring clostridium botulinum type a in rice samples.	a quantitative real-time pcr using sybr green dye was developed to target the neurotoxin type a (bont/a) gene of clostridium botulinum type a. primer specificity was confirmed by analyzing 63 strains including 5 strains of c. botulinum type a and 11 of non-type a c. botulinum. the highly similar amplification efficiencies of the real-time pcr assay were observed for 5 strains of c. botulinum type a. the dna extraction with nuclisens minimag provided sufficient performance to obtain the purified ...	2010	20377957
c3 peptide enhances recovery from spinal cord injury by improved regenerative growth of descending fiber tracts.	functional recovery and regeneration of corticospinal tract (cst) fibers following spinal cord injury by compression or dorsal hemisection in mice was monitored after application of the enzyme-deficient clostridium botulinum c3-protein-derived 29-amino-acid fragment c3bot(154-182). this peptide significantly improved locomotor restoration in both injury models as assessed by the open-field basso mouse scale for locomotion test and rotarod treadmill experiments. these data were supported by traci ...	2010	20406886
optimization of the bont/a-hc expression in recombinant escherichia coli using the taguchi statistical method.	the use of the recombinant bont/a-hc (carboxylic domain of the clostridium botulinum neurotoxin heavy chain) has been proposed as a vaccine candidate for botulism. this fragment contains the principle protective antigenic determinant. in the present study, in order to maximize recombinant protein expression, after verification of recombinant bont/a-hc by western blotting, modified m9 medium was selected as a simple medium, and the operational and medium-composition variables together with their ...	2010	20412050
botulinum toxin.	botulinum toxin, one of the most poisonous biological substances known, is a neurotoxin produced by the bacterium clostridium botulinum. c. botulinum elaborates eight antigenically distinguishable exotoxins (a, b, c(1), c(2), d, e, f and g). all serotypes interfere with neural transmission by blocking the release of acetylcholine, the principal neurotransmitter at the neuromuscular junction, causing muscle paralysis. the weakness induced by injection with botulinum toxin a usually lasts about th ...	2010	20418969
pentaplexed quantitative real-time pcr assay for the simultaneous detection and quantification of botulinum neurotoxin-producing clostridia in food and clinical samples.	botulinum neurotoxins are produced by the anaerobic bacterium clostridium botulinum and are divided into seven distinct serotypes (a to g) known to cause botulism in animals and humans. in this study, a multiplexed quantitative real-time pcr assay for the simultaneous detection of the human pathogenic c. botulinum serotypes a, b, e, and f was developed. based on the taqman chemistry, we used five individual primer-probe sets within one pcr, combining both minor groove binder- and locked nucleic ...	2010	20435756
rapid affinity immunochromatography column-based tests for sensitive detection of clostridium botulinum neurotoxins and escherichia coli o157.	existing methods for detection of food-borne pathogens and their toxins are frequently time-consuming, require specialized equipment, and involve lengthy culture procedures and/or animal testing and are thus unsuitable for a rapid response to an emergency public health situation. a series of simple and rapid affinity immunochromatography column (aicc) assays were developed to detect clostridium botulinum neurotoxin types a, b, e, and f and escherichia coli o157 in food matrices. specifically, fo ...	2010	20435757
identical novel a5(b3') botulinum neurotoxin gene arrangements isolated from widely disparate geographical and patient sources suggest their independent origins.		2010	20444983
preliminary x-ray crystallographic study of the receptor-binding domain of the d/c mosaic neurotoxin from clostridium botulinum.	botulinum toxin (bont) from clostridium botulinum ofd05, isolated from bovine botulism, is a d/c mosaic-type bont. bonts possess binding, translocation and catalytic domains. the bont/ofd05 binding domain exhibits significant sequence identity to bont/c, which requires a single ganglioside as a binding receptor on neuronal cells, while bont/a and bont/b require two receptors for specific binding. to determine the binding mechanism of bont/ofd05 and its ganglioside receptors on neuronal cells, re ...	2010	20445271
regulation of neurotoxin production and sporulation by a putative agrbd signaling system in proteolytic clostridium botulinum.	a significant number of genome sequences of clostridium botulinum and related species have now been determined. in silico analysis of these data revealed the presence of two distinct agr loci (agr-1 and agr-2) in all group i strains, each encoding putative proteins with similarity to agrb and agrd of the well-studied staphylococcus aureus agr quorum sensing system. in s. aureus, a small diffusible autoinducing peptide is generated from agrd in a membrane-located processing event that requires ag ...	2010	20453132
involvement of sialic acid in transport of serotype c1 botulinum toxins through rat intestinal epithelial cells.	clostridium botulinum produces a large toxin complex (l-tc) composed of neurotoxin (bont) and non-toxic proteins. in animal botulism, bont or l-tc is absorbed via the intestinal epithelium. to establish the cellular mechanisms of botulinum toxin absorption, we used cultured rat intestinal epithelial cells to test the binding and transport of serotype c1 bont and l-tc through the cell layers. bont and l-tc bound to and passed through the cell layers, with l-tc exhibiting larger binding and transp ...	2010	20453451
botulinum hemagglutinin disrupts the intercellular epithelial barrier by directly binding e-cadherin.	botulinum neurotoxin is produced by clostridium botulinum and forms large protein complexes through associations with nontoxic components. we recently found that hemagglutinin (ha), one of the nontoxic components, disrupts the intercellular epithelial barrier; however, the mechanism underlying this phenomenon is not known. in this study, we identified epithelial cadherin (e-cadherin) as a target molecule for ha. ha directly binds e-cadherin and disrupts e-cadherin-mediated cell to cell adhesion. ...	2010	20457762
re-engineering clostridial neurotoxins for the treatment of chronic pain: current status and future prospects.	clostridial neurotoxins from the botulinum neurotoxin (bont) family are protein complexes, derived from the bacterium clostridium botulinum, which potently inhibit acetylcholine release and result in a reversible blockade of the neuromuscular junction. this feature led to the clinical development of bont-a for a number of neuromuscular disorders. bont-a toxins are commercially available as three different preparations: dysport/azzalure, botox/vistabel, and xeomin/bocouture. although bont-a prepa ...	2010	20462283
clostridium botulinum in the post-genomic era.	foodborne botulism is a severe neuroparalytic disease caused by consumption of botulinum neurotoxin formed by strains of proteolytic clostridium botulinum and non-proteolytic c. botulinum during their growth in food. the botulinum neurotoxin is the most potent substance known, with as little as 30-100 ng potentially fatal, and consumption of just a few milligrams of neurotoxin-containing food is likely to be sufficient to cause illness and potentially death. in order to minimise the foodborne bo ...	2010	21315972
challenges in risk assessment and predictive microbiology of foodborne spore-forming bacteria.	mathematical description of the behavior of bacterial foodborne pathogens and concepts of risk assessment were first applied to spore-forming bacteria and specially to clostridium botulinum with numerous works dealing with spores heat destruction to ensure the safety of canned foods or with their germination and growth probability in foods. this paper discusses two aspects which appear specific to pathogenic sporeformers in comparison to vegetative microorganisms, that is, firstly, the extreme i ...	2010	21315975
lag time variability in individual spores of clostridium botulinum.	quantifying lag times from individual spores and the associated variability is an important part of understanding the hazard associated with spore-forming pathogens such as clostridium botulinum. knowledge of the underlying distribution would allow greater refinement of risk assessments. to date most studies have either examined lag time indirectly by measuring time to growth or have only examined the first stage of lag, germination. recent studies have attempted to quantify the variability of s ...	2010	21315978
quantitative risk assessment for hazards that arise from non-proteolytic clostridium botulinum in minimally processed chilled dairy-based foods.	a modular process risk model has been constructed that describes the manufacture of dairy dessert products and hazards that arise from non-proteolytic clostridium botulinum. the model describes batch manufacture and consumer storage of a family size generic dairy dessert but includes a realistic quantification that could apply to a specific food product. the dairy dessert sector is an expanding part of the uk market. the model includes modules that describe spore loads in raw materials, spore in ...	2010	21315990
edaphic and phytochemical factors as predictors of equine grass sickness cases in the uk.	background: equine dysautonomia or equine grass sickness (egs), as it is more commonly known, is a usually fatal disease of equids of uncertain etiology, although associated with grazing, that affects the autonomic and enteric nervous system. lowered gastrointestinal motility, leading to paralysis of the gut, is one of the main symptoms of egs. previous studies have implicated anaerobic bacteria, notably clostridium botulinum, but what triggers the severe bacterial infestations remains enigmatic ...	2010	21833167
botulism in brazil, 2000-2008: epidemiology, clinical findings and laboratorial diagnosis.	botulism is a rare and potentially lethal illness caused by clostridium botulinum neurotoxin. we describe the findings of a laboratorial investigation of 117 suspected cases of botulism reported to the surveillance system in brazil from january 2000 to october 2008. data on the number and type of samples analyzed, type of toxins identified, reporting of the number of botulism cases and transmission sources are discussed. a total of 193 clinical samples and 81 food samples were analyzed for detec ...	2010	21748224
molecular characterization and comparison of clostridium botulinum type c avian strains.	type c botulinum neurotoxin (bont/c)-producing clostridium botulinum causes animal botulism worldwide and has become a serious problem in poultry flocks and waterfowl in sweden. the objectives of the present study were to isolate, characterize and subtype c. botulinum type c avian isolates in order to increase the knowledge of the genetic diversity. isolates from 13 birds were identified by 16s rrna sequencing and bont/c gene detection by real-time polymerase chain reaction (pcr). conventional p ...	2010	21154062
images in clinical medicine: wound botulism.		2010	21158660
different substrate recognition requirements for cleavage of synaptobrevin-2 by clostridium baratii and clostridium botulinum type f neurotoxins.	botulinum neurotoxins (bonts) cause botulism, which can be fatal if it is untreated. bonts cleave proteins necessary for nerve transmission, resulting in paralysis. the in vivo protein target has been reported for all seven serotypes of bont, i.e., serotypes a to g. knowledge of the cleavage sites has led to the development of several assays to detect bont based on its ability to cleave a peptide substrate derived from its in vivo protein target. most serotypes of bont can be subdivided into sub ...	2010	21169446
botulism, where are we now?	botulism is a neuroparalytic illness caused by botulinum toxin, a product of the clostridium botulinum bacteria and characteristically presents as an acute, symmetrical, descending flaccid paralysis. albeit it is the most poisonous substance known, which even poses a major threat as biological weapons, purified and highly diluted botulinum toxin can be used to treat a wide variety of conditions associated with muscular hyperactivity, glandular hypersecretions and pain. there are six clinical pre ...	2010	21171845
type e botulism.	there are seven known serotypes of botulism, designated a through g; almost all human cases of botulism are caused by types a, b, and e. botulism type e is the predominant serotype causing disease associated with native arctic foods. in the circumpolar regions of the world, the coastal soils are rich in botulism type e, and consumption of fish and marine animals in these areas are the sources of clusters of botulism. unlike spores of type a and b, botulism type e can withstand freezing down to 3 ...	2010	21171846
analysis of genomic differences among clostridium botulinum type a1 strains.	type a1 clostridium botulinum strains are a group of gram-positive, spore-forming anaerobic bacteria that produce a genetically, biochemically, and biophysically indistinguishable 150 kd protein that causes botulism. the genomes of three type a1 c. botulinum strains have been sequenced and show a high degree of synteny. the purpose of this study was to characterize differences among these genomes and compare these differentiating features with two additional unsequenced strains used in previous ...	2010	21182778
novel structural elements within the nonproteolytic clostridium botulinum type f toxin gene cluster.	we sequenced for the first time the complete neurotoxin gene cluster of a nonproteolytic clostridium botulinum type f. the neurotoxin gene cluster contained a novel gene arrangement that, compared to other c. botulinum neurotoxin gene clusters, lacked the regulatory botr gene and contained an intergenic is element between its orfx2 and orfx3 genes.	2010	21183631
therapeutic effects of clostridium botulinum c3 exoenzyme.	c3 exoenzyme from clostridium botulinum, specifically adp-ribosylates small gtp-binding proteins rhoa, b, and c. adp-ribosylation causes functional inactivation of rho proteins resulting in cessation of the complete downstream signaling. rho proteins are general regulators of a lot of essential cellular functions, among others, the neuronal growth cone. rho activation, triggered by neuronal injury, inhibits neuronal repair mechanisms. to prevent the detrimental effect of active rho in the recove ...	2010	21193903
studies on the dissociation of botulinum neurotoxin type a complexes.	the neurotoxins produced by the various strains of the anaerobic bacterium clostridium botulinum naturally occur associated with complexing proteins which serve to protect the neurotoxins from the harsh environment of the mammalian gastrointestinal tract during bacterial invasion of the host. three different complex species with the discrete sizes 19s (900 kda, ll complex), 16s (500 kda, l complex) and 12s (300 kda, m complex) may be isolated from c. botulinum type a cultures. however, to affect ...	2010	21195107
structure and activity of a functional derivative of clostridium botulinum neurotoxin b.	botulinum neurotoxins (bonts) cause flaccid paralysis by inhibiting neurotransmission at cholinergic nerve terminals. bonts consist of three essential domains for toxicity: the cell binding domain (hc), the translocation domain (hn) and the catalytic domain (lc). a functional derivative (lhn) of the parent neurotoxin b composed of hn and lc domains was recombinantly produced and characterised. lhn/b crystallographic structure at 2.8å resolution is reported. the catalytic activity of lhn/b toward ...	2010	21078393
prophylactic botulinum type a toxin complex (dysport®) for migraine without aura.	to evaluate the efficacy, safety, and optimum dose of a highly purified clostridium botulinum type a toxin-hemagglutinin complex (dysport) for migraine prophylaxis.	2010	21083558
[quick recovery, after administration of a botulinic antitoxin serum in two patients presenting with botulism].		2010	21094008
on the role of rhoa/rock signaling in contact guidance of bone-forming cells on anisotropic ti6al4v surfaces.	patterned surfaces direct cell spatial dynamics, yielding cells oriented along the surface geometry, in a process known as contact guidance. the rho family of gtpases controls the assembly of focal adhesions and cytoskeleton dynamics, but its role in modulating bone-cell alignment on patterned surfaces remains unknown. this article describes the interactions of two human cell types involved in osseointegration, specifically mesenchymal stem cells and osteoblasts, with submicron- or nano-scale ti ...	2010	21115140
biodiversity of clostridium botulinum type e associated with a large outbreak of botulism in wildlife from lake erie and lake ontario.	the genetic relatedness of clostridium botulinum type e isolates associated with an outbreak of wildlife botulism was studied using random amplification of polymorphic dna (rapd). specimens were collected from november 2000 to december 2008 during a large outbreak of botulism affecting birds and fish living in and around lake erie and lake ontario. in our present study, a total of 355 wildlife samples were tested for the presence of botulinum toxin and/or organisms. type e botulinum toxin was de ...	2010	21115703
exploring the role of host cell chaperones/ppiases during cellular up-take of bacterial adp-ribosylating toxins as basis for novel pharmacological strategies to protect mammalian cells against these virulence factors.	bacterial exotoxins exploit protein transport pathways of their mammalian target cells to deliver their enzymatic active moieties into the cytosol. there, they modify their specific substrate molecules resulting in cell damage and the clinical symptoms characteristic for each individual toxin. we have investigated the cellular uptake of the binary actin adp-ribosylating c2 toxin from clostridium botulinum and the binary lethal toxin from bacillus anthracis, a metalloprotease. both toxins are com ...	2010	21120455
crystal structure of the receptor binding domain of the botulinum c-d mosaic neurotoxin reveals potential roles of lysines 1118 and 1136 in membrane interactions.	the botulinum neurotoxins (bonts) produced by different strains of the bacterium clostridium botulinum are responsible for the disease botulism and include a group of immunologically distinct serotypes (a, b, e, and f) that are considered to be the most lethal natural proteins known for humans. two bont serotypes, c and d, while rarely associated with human infection, are responsible for deadly botulism outbreaks afflicting animals. also associated with animal infections is the bont c-d mosaic p ...	2010	21130733
from cosubstrate similarity to inhibitor diversity--inhibitors of adp-ribosyltransferases from kinase inhibitor screening.	adp-ribosyltransferases (adp-rts) use nad(+) to transfer an adp-ribosyl group to target proteins. although some adp-rts are bacterial toxins only few inhibitors are known. here we present the development of fluorescence-based assays and a focussed library screening using kinase inhibitors as a new approach towards inhibitors of adp-rts. different screening setups were established using surrogate small molecule substrates or the quantitation of the cofactor nad(+). proof-of-principle screening ex ...	2010	21132207
a label-free biosensor assay for botulinum neurotoxin b in food and human serum.	botulinum neurotoxins (bonts) are among the most toxic substances known. surveillance and diagnostics require methods for rapid detection of bonts in complex media such as foodstuffs and human serum. we have developed in vitro assays to specifically detect the protease activity of botulinum neurotoxin b (bont/b) on a time scale of minutes. cleavage of the bont/b substrate vamp2, a membrane snare protein associated with synaptic vesicles, was monitored using real-time surface plasmon resonance to ...	2010	21134347
internalization of biotinylated compounds into cancer cells is promoted by a molecular trojan horse based upon core streptavidin and clostridial c2 toxin.	the c2 toxin produced by clostridium botulinum is a binary ab-type exotoxin composed of the enzyme subunit c2i and the binding/translocation moiety c2ii. after proteolytic activation, c2iia mediates the subsequent internalization of c2i into the cytosol of mammalian target cells. the n-terminal domain of c2i (c2in) is necessary for c2iia-dependent uptake, but lacks the enzyme domain that is responsible for cytotoxicity. in the present study, we generated a delivery system building on c2in and a ...	2010	21136248
a dileucine in the protease of botulinum toxin a underlies its long-lived neuroparalysis: transfer of longevity to a novel potential therapeutic.	blockade of neurotransmitter release by botulinum neurotoxin type a (bont(a)) underlies the severe neuroparalytic symptoms of human botulism, which can last a few years. the structural basis for this remarkable persistence remains unclear. herein, recombinant bont(a) was found to match the neurotoxicity of that from clostridium botulinum, producing persistent cleavage of synaptosomal-associated protein of 25 kda (snap-25) and neuromuscular paralysis. when two leucines near the c terminus of the ...	2010	21138836
high-level expression, purification, crystallization and preliminary x-ray crystallographic studies of the receptor-binding domain of botulinum neurotoxin serotype d.	botulinum neurotoxins (bonts) are highly toxic proteins for humans and animals that are responsible for the deadly neuroparalytic disease botulism. here, details of the expression and purification of the receptor-binding domain (hcr) of bont/d in escherichia coli are presented. using a codon-optimized cdna, bont/d_hcr was expressed at a high level (150-200 mg per litre of culture) in the soluble fraction. following a three-step purification protocol, very pure (>98%) bont/d_hcr was obtained. the ...	2010	21139207
botulinum toxin: bioweapon & magic drug.	botulinum neurotoxins, causative agents of botulism in humans, are produced by clostridium botulinum, an anaerobic spore-former gram positive bacillus. botulinum neurotoxin poses a major bioweapon threat because of its extreme potency and lethality; its ease of production, transport, and misuse; and the need for prolonged intensive care among affected persons. a single gram of crystalline toxin, evenly dispersed and inhaled, can kill more than one million people. the basis of the phenomenal pote ...	2010	21149997
a tracing tool portfolio to detect bacillus anthracis, clostridium botulinum and noroviruses: bioterrorism is a food safety and security issue.		2010	21324542
protein domain analysis of c. botulinum type a neurotoxin and its relationship with other botulinum serotypes.	botulinum neurotoxins (bonts) are highly potent poisons produced by seven serotypes of clostridium botulinum. the mechanism of neurotoxin action is a multistep process which leads to the cleavage of one of three different snare proteins essential for synaptic vesicle fusion and transmission of the nerve signals to muscles: synaptobrevin, syntaxin, or snap-25. in order to understand the precise mechanism of neurotoxin in a host, the domain structure of the neurotoxin was analyzed among different ...	2010	22069543
the mechanism by which rhoa regulates vascular reactivity after hemorrhagic shock in rats.	rhoa, an important member of the rho family of gtpases, has been implicated in many cellular processes. our pilot study found that rhoa participated in the regulation of vascular reactivity after shock, but the mechanism was incompletely understood. whether rhoa regulates vascular reactivity through the rho kinase-myosin light-chain phosphatase (mlcp) and rac1-p21-activated kinase (pak)-myosin light-chain kinase (mlck) signaling pathway needs investigation. with isolated, superior mesenteric art ...	2010	20472763
infant botulism.	infant botulism is a rare condition caused by intestinal colonization with clostridium botulinum. the enteric toxin causes intestinal immobility and progressive descending paralysis due to the effect on acetylcholine release at the neuromuscular junction and other cholinergic nerve terminals, particularly in the gut. herein, we report an infant with infantile botulism because of rare clinically entity, with early diagnosis and appropriate treatment recover no squeal.	2010	25610131
the hazards of honey: infantile botulism.	infantile botulism is a rare cause of neuromuscular weakness resulting from ingestion of clostridium botulinum-an anaerobic gram-positive bacillus found universally in soil. the only definite food source known to cause infantile botulism is honey; previously, links to formula milk have been postulated but not definitely sourced. we present an interesting case report of a 2-month-old infant with this rare condition, including the diagnostic difficulties that ensued. a brief overview of the condit ...	2010	22778374
botulinum toxin: therapeutic agent to cosmetic enhancement to lethal biothreat.	the clinical effects of clostridium botulinum and its extremely potent neurotoxin have been known for two centuries. the disease threat and the clinical uses are now well established. what's changed is the potential for botulinum neurotoxin to be used as a biological threat agent. the recent upsurge of illegal trafficking of reagent-grade toxin could, if bought in large enough quantities, be as serious a threat as other biothreat agents such as anthrax and smallpox, which have received much more ...	2011	22178572
Two outbreaks of botulism associated with consumption of green olive paste, France, September 2011.		2011	22172329
role of rho gtpases in human trophoblast migration induced by igfbp1.	insulin like growth factor binding protein-1 (igfbp1), the main secretory product of the decidualized endometrium of a pregnant woman has previously been shown to interact with the alpha5beta1 integrin of evt cell surface to stimulate its migration in an igf-independent manner. this migration stimulation has also been shown to require activation of extracellular signal regulated kinases 1 and 2 (mapk3/1) and focal adhesion kinase (fak, official symbol ptk). the present study examined the roles o ...	2011	21940708