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Sample records for botulinum neurotoxin type

  1. Botulinum Neurotoxin Type A in Neurology: Update

    OpenAIRE

    Orsini, Marco; Leite, Marco Antonio Araujo; Chung, Tae Mo; Bocca, Wladimir; de Souza, Jano Alves; de Souza, Olivia Gameiro; Moreira, Rayele Priscila; Bastos, Victor Hugo; Teixeira, Silmar; Oliveira, Acary Bulle; Moraes, Bruno da Silva; Matta, André Palma; Jacinto, Luis Jorge

    2015-01-01

    This paper reviews the current and most neurological (central nervous system, CNS) uses of the botulinum neurotoxin type A. The effect of these toxins at neuromuscular junction lends themselves to neurological diseases of muscle overactivity, particularly abnormalities of muscle control. There are seven serotypes of the toxin, each with a specific activity at the molecular level. Currently, serotypes A (in two preparations) and B are available for clinical purpose, and they have proved to be ...

  2. Neutralization of Botulinum Neurotoxin Type E by a Humanized Antibody

    Science.gov (United States)

    Derman, Yağmur; Selby, Katja; Miethe, Sebastian; Frenzel, André; Liu, Yvonne; Rasetti-Escargueil, Christine; Avril, Arnaud; Pelat, Thibaut; Urbain, Remi; Fontayne, Alexandre; Thullier, Philippe; Sesardic, Dorothea; Lindström, Miia; Hust, Michael; Korkeala, Hannu

    2016-01-01

    Botulinum neurotoxins (BoNTs) cause botulism and are the deadliest naturally-occurring substances known to humans. BoNTs have been classified as one of the category A agents by the Centers for Disease Control and Prevention, indicating their potential use as bioweapons. To counter bio-threat and naturally-occurring botulism cases, well-tolerated antibodies by humans that neutralize BoNTs are relevant. In our previous work, we showed the neutralizing potential of macaque (Macaca fascicularis)-derived scFv-Fc (scFv-Fc ELC18) by in vitro endopeptidase immunoassay and ex vivo mouse phrenic nerve-hemidiaphragm assay by targeting the light chain of the botulinum neurotoxin type E (BoNT/E). In the present study, we germline-humanized scFv-Fc ELC18 into a full IgG hu8ELC18 to increase its immunotolerance by humans. We demonstrated the protection and prophylaxis capacity of hu8ELC18 against BoNT/E in a mouse model. A concentration of 2.5 ng/mouse of hu8ELC18 protected against 5 mouse lethal dose (MLD) in a mouse protection assay and complete neutralization of 1 LD50 of pure BoNT/E toxin was achieved with 8 ng of hu8ELC18 in mouse paralysis assay. Furthermore, hu8ELC18 protected mice from 5 MLD if injected up to 14 days prior to intraperitoneal BoNT/E administration. This newly-developed humanized IgG is expected to have high tolerance in humans. PMID:27626446

  3. Neutralization of Botulinum Neurotoxin Type E by a Humanized Antibody

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    Yağmur Derman

    2016-09-01

    Full Text Available Botulinum neurotoxins (BoNTs cause botulism and are the deadliest naturally-occurring substances known to humans. BoNTs have been classified as one of the category A agents by the Centers for Disease Control and Prevention, indicating their potential use as bioweapons. To counter bio-threat and naturally-occurring botulism cases, well-tolerated antibodies by humans that neutralize BoNTs are relevant. In our previous work, we showed the neutralizing potential of macaque (Macaca fascicularis-derived scFv-Fc (scFv-Fc ELC18 by in vitro endopeptidase immunoassay and ex vivo mouse phrenic nerve-hemidiaphragm assay by targeting the light chain of the botulinum neurotoxin type E (BoNT/E. In the present study, we germline-humanized scFv-Fc ELC18 into a full IgG hu8ELC18 to increase its immunotolerance by humans. We demonstrated the protection and prophylaxis capacity of hu8ELC18 against BoNT/E in a mouse model. A concentration of 2.5 ng/mouse of hu8ELC18 protected against 5 mouse lethal dose (MLD in a mouse protection assay and complete neutralization of 1 LD50 of pure BoNT/E toxin was achieved with 8 ng of hu8ELC18 in mouse paralysis assay. Furthermore, hu8ELC18 protected mice from 5 MLD if injected up to 14 days prior to intraperitoneal BoNT/E administration. This newly-developed humanized IgG is expected to have high tolerance in humans.

  4. Sensitive Detection of Botulinum Neurotoxin Types C and D with an Immunoaffinity Chromatographic Column Test

    OpenAIRE

    Gessler, Frank; Hampe, Katrin; Böhnel, Helge

    2005-01-01

    A sensitive and specific immunoassay for the simultaneous detection of Clostridium botulinum type C (BoNT/C) and type D neurotoxin was developed. Goat anti-mouse immunoglobulin G was bound to polyethylene disks in a small disposable column used for this assay. The sample was preincubated together with monoclonal antibodies specific for the heavy chain of BoNT/C and D and affinity-purified, biotinylated polyclonal antibodies against these neurotoxins. This complex was captured on the assay dis...

  5. Analysis of a unique Clostridium botulinum strain from the Southern hemisphere producing a novel type E botulinum neurotoxin subtype

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    Raphael Brian H

    2012-10-01

    Full Text Available Abstract Background Clostridium botulinum strains that produce botulinum neurotoxin type E (BoNT/E are most commonly isolated from botulism cases, marine environments, and animals in regions of high latitude in the Northern hemisphere. A strain of C. botulinum type E (CDC66177 was isolated from soil in Chubut, Argentina. Previous studies showed that the amino acid sequences of BoNT/E produced by various strains differ by rarA operon. Results Genetic and mass spectral analysis demonstrated that the BoNT/E produced by CDC66177 is a novel toxin subtype (E9. Toxin gene sequencing indicated that BoNT/E9 differed by nearly 11% at the amino acid level compared to BoNT/E1. Mass spectrometric analysis of BoNT/E9 revealed that its endopeptidase substrate cleavage site was identical to other BoNT/E subtypes. Further analysis of this strain demonstrated that its 16S rRNA sequence clustered with other Group II C. botulinum (producing BoNT types B, E, and F strains. Genomic DNA isolated from strain CDC66177 hybridized with fewer probes using a Group II C. botulinum subtyping microarray compared to other type E strains examined. Whole genome shotgun sequencing of strain CDC66177 revealed that while the toxin gene cluster inserted into the rarA operon similar to other type E strains, its overall genome content shared greater similarity with a Group II C. botulinum type B strain (17B. Conclusions These results expand our understanding of the global distribution of C. botulinum type E strains and suggest that the type E toxin gene cluster may be able to insert into C. botulinum strains with a more diverse genetic background than previously recognized.

  6. Clinical differences between botulinum neurotoxin type A and B.

    Science.gov (United States)

    Bentivoglio, Anna Rita; Del Grande, Alessandra; Petracca, Martina; Ialongo, Tamara; Ricciardi, Lucia

    2015-12-01

    In humans, the therapeutic use of botulinum neurotoxin A (BoNT/A) is well recognized and continuously expanding. Four BoNTs are widely available for clinical practice: three are serotype A and one is serotype B: onabotulinumtoxinA (A/Ona), abobotulinumtoxinA (A/Abo) and incobotulinumtoxinA (A/Inco), rimabotulinumtoxinB (B/Rima). A/Abo, A/Inco, A/Ona and B/Rima are all licensed worldwide for cervical dystonia. In addition, the three BoNT/A products are approved for blepharospasm and focal dystonias, spasticity, hemifacial spasm, hyperhidrosis and facial lines, with remarkable regional differences. These toxin brands differ for specific activity, packaging, constituents, excipient, and storage. Comparative literature assessing the relative safety and efficacy of different BoNT products is limited, most data come from reports on small samples, and only a few studies meet criteria of evidence-based medicine. One study compared the effects of BoNT/A and BoNT/B on muscle activity of healthy volunteers, showing similar neurophysiological effects with a dose ratio of 1:100. In cervical dystonia, when comparing the effects of BoNT/A and BoNT/B, results are more variable, some studies reporting roughly similar peak effect and overall duration (at a ratio of 1:66, others reporting substantially shorter duration of BoNT/B than BoNT/A (at a ratio 1/24). Although the results of clinical studies are difficult to compare for methodological differences (dose ratio, study design, outcome measures), it is widely accepted that: BoNT/B is clinically effective using appropriate doses as BoNT/A (1:40-50), injections are generally more painful, in most of the studies on muscular conditions, efficacy is shorter, and immunogenicity higher. Since the earliest clinical trials, it has been reported that autonomic side effects are more frequent after BoNT/B injections, and this observation encouraged the use of BoNT/B for sialorrhea, hyperhidrosis and other non-motor symptoms. In these

  7. Purification, Potency, and Efficacy of the Botulinum Neurotoxin Type A Binding Domain from Pichia pastoris as a Recombinant Vaccine Candidate

    OpenAIRE

    Byrne, Michael P.; Smith, Theresa J.; Montgomery, Vicki A.; Smith, Leonard A

    1998-01-01

    Recombinant botulinum neurotoxin serotype A binding domain [BoNT/A(Hc)], expressed in Pichia pastoris, was developed as a vaccine candidate for preventing botulinum neurotoxin type A (BoNT/A) intoxication. After fermentation and cell disruption, BoNT/A(Hc) was purified by using a three-step chromatographic process consisting of expanded-bed chromatography, Mono S cation-exchange chromatography, and hydrophobic interaction chromatography. Two pools of immunogenic product were separated on the ...

  8. Neutralizing antibodies to botulinum neurotoxin type A in aesthetic medicine: five case reports

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    Torres S

    2013-12-01

    Full Text Available Sebastian Torres,1 Mark Hamilton,2 Elena Sanches,4 Polina Starovatova,3 Elena Gubanova,3 Tatiana Reshetnikova51Di Stefano Velona Clinic, Catania, Italy; 2Hamilton Face Clinic, Dublin, Ireland; 3Preventive Medicine Clinic "Vallex M", Moscow, Russia; 4EKLAN Co Ltd Medical Center for Aesthetic Correction, Moscow, Russia; 5Department of Dermatovenereology and Cosmetology, State Medical University, Novosibirsk, RussiaAbstract: Botulinum neurotoxin injections are a valuable treatment modality for many therapeutic indications as well as in the aesthetic field for facial rejuvenation. As successful treatment requires repeated injections over a long period of time, secondary resistance to botulinum toxin preparations after repeated injections is an ongoing concern. We report five case studies in which neutralizing antibodies to botulinum toxin type A developed after injection for aesthetic use and resulted in secondary treatment failure. These results add to the growing number of reports in the literature for secondary treatment failure associated with high titers of neutralizing antibodies in the aesthetic field. Clinicians should be aware of this risk and implement injection protocols that minimize resistance development.Keywords: aesthetic medicine, botulinum neurotoxin type A, neutralizing antibody, secondary treatment failure

  9. Sensitive detection of botulinum neurotoxin types C and D with an immunoaffinity chromatographic column test.

    Science.gov (United States)

    Gessler, Frank; Hampe, Katrin; Böhnel, Helge

    2005-12-01

    A sensitive and specific immunoassay for the simultaneous detection of Clostridium botulinum type C (BoNT/C) and type D neurotoxin was developed. Goat anti-mouse immunoglobulin G was bound to polyethylene disks in a small disposable column used for this assay. The sample was preincubated together with monoclonal antibodies specific for the heavy chain of BoNT/C and D and affinity-purified, biotinylated polyclonal antibodies against these neurotoxins. This complex was captured on the assay disk. Streptavidin-poly-horseradish peroxidase was used as a conjugate, and a precipitating substrate allowed the direct semiquantitative readout of the assay, if necessary. For a more accurate quantitative detection, the substrate can be eluted and measured in a photometer. Depending on the preincubation time, a sensitivity of 1 mouse lethal dose ml(-1) was achieved in culture supernatants. PMID:16332765

  10. Botulinum neurotoxin type A modulates vesicular release of glutamate from satellite glial cells

    OpenAIRE

    da Silva, Larissa Bittencourt; Poulsen, Jeppe Nørgaard; Arendt-Nielsen, Lars; Gazerani, Parisa

    2015-01-01

    This study investigated the presence of cell membrane docking proteins synaptosomal-associated protein, 25 and 23 kD (SNAP-25 and SNAP-23) in satellite glial cells (SGCs) of rat trigeminal ganglion; whether cultured SGCs would release glutamate in a time- and calcium-dependent manner following calcium-ionophore ionomycin stimulation; and if botulinum neurotoxin type A (BoNTA), in a dose-dependent manner, could block or decrease vesicular release of glutamate. SGCs were isolated from the trige...

  11. PEG precipitation coupled with chromatography is a new and sufficient method for the purification of botulinum neurotoxin type B [corrected].

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    Yao Zhao

    Full Text Available Clostridium botulinum neurotoxins are used to treat a variety of neuro-muscular disorders, as well as in cosmetology. The increased demand requires efficient methods for the production and purification of these toxins. In this study, a new purification process was developed for purifying type B neurotoxin. The kinetics of C.botulinum strain growth and neurotoxin production were determined for maximum yield of toxin. The neurotoxin was purified by polyethylene glycol (PEG precipitation and chromatography. Based on design of full factorial experiment, 20% (w/v PEG-6000, 4 °C, pH 5.0 and 0.3 M NaCl were optimal conditions to obtain a high recovery rate of 87% for the type B neurotoxin complex, as indicated by a purification factor of 61.5 fold. Furthermore, residual bacterial cells, impurity proteins and some nucleic acids were removed by PEG precipitation. The following purification of neurotoxin was accomplished by two chromatography techniques using Sephacryl™ S-100 and phenyl HP columns. The neurotoxin was recovered with an overall yield of 21.5% and the purification factor increased to 216.7 fold. In addition, a mouse bioassay determined the purified neurotoxin complex possessed a specific toxicity (LD(50 of 4.095 ng/kg.

  12. Molecular Evolution of Antibody Cross-Reactivity for Two Subtypes of Type a Botulinum Neurotoxin

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Rodriguez, C.; Levy, R.; Arndt, J.W.; Forsyth, C.M.; Razai, A.; Lou, J.; Geren, I.; Stevens, R.C.; Marks, J.D.; /UC, San Francisco /Scripps Res. Inst.

    2007-07-09

    Broadening antibody specificity without compromising affinity should facilitate detection and neutralization of toxin and viral subtypes. We used yeast display and a co-selection strategy to increase cross-reactivity of a single chain (sc) Fv antibody to botulinum neurotoxin type A (BoNT/A). Starting with a scFv that binds the BoNT/A1 subtype with high affinity (136 pM) and the BoNT/A2 subtype with low affinity (109 nM), we increased its affinity for BoNT/A2 1,250-fold, to 87 pM, while maintaining high-affinity binding to BoNT/A1 (115 pM). To find the molecular basis for improved cross-reactivity, we determined the X-ray co-crystal structures of wild-type and cross-reactive antibodies complexed to BoNT/A1 at resolutions up to 2.6 A, and measured the thermodynamic contribution of BoNT/A1 and A2 amino acids to wild-type and cross-reactive antibody binding. The results show how an antibody can be engineered to bind two different antigens despite structural differences in the antigen-antibody interface and may provide a general strategy for tuning antibody specificity and cross-reactivity.

  13. Botulinum neurotoxin type A versus punctal plug insertion in the management of dry eye disease

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    Amal A Bukhari

    2014-01-01

    Full Text Available Purpose: To compare the efficacies of punctal plug insertion and Botulinum toxin injection in dry eye disease not responding to topical medications. Materials and Methods: A non-controlled randomized clinical trial of two parallel groups of 60 dry eye patients seen in the clinic not responding to topical medications were divided into two groups. One group received punctal plugs and the other group received Botulinum toxin injections to prevent lacrimal tear drainage. Results: Of a total of 36 patients with a mean age of 44.5 years who received punctal plugs, 50% of them experienced improvements in the clinical manifestations of their disease. 12/36 (33.3% developed plug extrusion, and 6/36 (16.7% patients developed conjunctival erosions with irritation that necessitated plug removal within one week of insertion. A total of 24 patients with a mean age of 47.5 years received injections of Botulinum toxin. Of these, 83.3% had improvement in all of the clinical manifestations of dry eye. 4/24 (16.7% had no improvement in the degrees to which they experienced foreign body sensations, 33.3% reported shampoo entering the eye while showering. All of the patients who received Botulinum toxin injections were satisfied with the results of their treatment, whereas only 72.3% of the patients who received punctal plugs were satisfied with their results. Conclusion: Botulinum neurotoxin A injections can be a very good alternative to punctal plugs in improving the clinical manifestations of dry eye disease They are associated with the development of fewer and milder complications and with higher levels of patient satisfaction.

  14. Zebrafish Sensitivity to Botulinum Neurotoxins

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    Kamalakar Chatla

    2016-05-01

    Full Text Available Botulinum neurotoxins (BoNT are the most potent known toxins. The mouse LD50 assay is the gold standard for testing BoNT potency, but is not sensitive enough to detect the extremely low levels of neurotoxin that may be present in the serum of sensitive animal species that are showing the effects of BoNT toxicity, such as channel catfish affected by visceral toxicosis of catfish. Since zebrafish are an important animal model for diverse biomedical and basic research, they are readily available and have defined genetic lines that facilitate reproducibility. This makes them attractive for use as an alternative bioassay organism. The utility of zebrafish as a bioassay model organism for BoNT was investigated. The 96 h median immobilizing doses of BoNT/A, BoNT/C, BoNT/E, and BoNT/F for adult male Tübingen strain zebrafish (0.32 g mean weight at 25 °C were 16.31, 124.6, 4.7, and 0.61 picograms (pg/fish, respectively. These findings support the use of the zebrafish-based bioassays for evaluating the presence of BoNT/A, BoNT/E, and BoNT/F. Evaluating the basis of the relatively high resistance of zebrafish to BoNT/C and the extreme sensitivity to BoNT/F may reveal unique functional patterns to the action of these neurotoxins.

  15. Botulinum neurotoxin type A induces TLR2-mediated inflammatory responses in macrophages.

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    Yun Jeong Kim

    Full Text Available Botulinum neurotoxin type A (BoNT/A is the most potent protein toxin and causes fatal flaccid muscle paralysis by blocking neurotransmission. Application of BoNT/A has been extended to the fields of therapeutics and biodefense. Nevertheless, the global response of host immune cells to authentic BoNT/A has not been reported. Employing microarray analysis, we performed global transcriptional profiling of RAW264.7 cells, a murine alveolar macrophage cell line. We identified 70 genes that were modulated following 1 nM BoNT/A treatment. The altered genes were mainly involved in signal transduction, immunity and defense, protein metabolism and modification, neuronal activities, intracellular protein trafficking, and muscle contraction. Microarray data were validated with real-time RT-PCR for seven selected genes including tlr2, tnf, inos, ccl4, slpi, stx11, and irg1. Proinflammatory mediators such as nitric oxide (NO and tumor necrosis factor alpha (TNFα were induced in a dose-dependent manner in BoNT/A-stimulated RAW264.7 cells. Increased expression of these factors was inhibited by monoclonal anti-Toll-like receptor 2 (TLR2 and inhibitors specific to intracellular proteins such as c-Jun N-terminal kinase (JNK, extracellular signal-regulated kinase (ERK, and p38 mitogen-activated protein kinase (MAPK. BoNT/A also suppressed lipopolysaccharide-induced NO and TNFα production from RAW264.7 macrophages at the transcription level by blocking activation of JNK, ERK, and p38 MAPK. As confirmed by TLR2-/- knock out experiments, these results suggest that BoNT/A induces global gene expression changes in host immune cells and that host responses to BoNT/A proceed through a TLR2-dependent pathway, which is modulated by JNK, ERK, and p38 MAPK.

  16. A historical and proteomic analysis of botulinum neurotoxin type/G

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    Rees Jon

    2011-10-01

    Full Text Available Abstract Background Clostridium botulinum is the taxonomic designation for at least six diverse species that produce botulinum neurotoxins (BoNTs. There are seven known serotypes of BoNTs (/A through/G, all of which are potent toxins classified as category A bioterrorism agents. BoNT/G is the least studied of the seven serotypes. In an effort to further characterize the holotoxin and neurotoxin-associated proteins (NAPs, we conducted an in silico and proteomic analysis of commercial BoNT/G complex. We describe the relative quantification of the proteins present in the/G complex and confirm our ability to detect the toxin activity in vitro. In addition, we review previous literature to provide a complete description of the BoNT/G complex. Results An in-depth comparison of protein sequences indicated that BoNT/G shares the most sequence similarity with the/B serotype. A temperature-modified Endopep-MS activity assay was successful in the detection of BoNT/G activity. Gel electrophoresis and in gel digestions, followed by MS/MS analysis of/G complex, revealed the presence of four proteins in the complexes: neurotoxin (BoNT and three NAPs--nontoxic-nonhemagglutinin (NTNH and two hemagglutinins (HA70 and HA17. Rapid high-temperature in-solution tryptic digestions, coupled with MS/MS analysis, generated higher than previously reported sequence coverages for all proteins associated with the complex: BoNT 66%, NTNH 57%, HA70 91%, and HA17 99%. Label-free relative quantification determined that the complex contains 30% BoNT, 38% NTNH, 28% HA70, and 4% HA17 by weight comparison and 17% BoNT, 23% NTNH, 42% HA70, and 17% HA17 by molecular comparison. Conclusions The in silico protein sequence comparisons established that the/G complex is phenetically related to the other six serotypes of C. botulinum. Proteomic analyses and Endopep-MS confirmed the presence of BoNT and NAPs, along with the activity of the commercial/G complex. The use of data

  17. CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN SEROTYPE B.

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    SWAMINATHAN,S.; ESWARAMOORTHY,S.

    2001-11-19

    The toxigenic strains of Clostridium botulinum produce seven serologically distinct types of neurotoxins labeled A - G (EC 3.4.24.69), while Clostridium tetani produces tetanus neurotoxin (EC 3.4.24.68). Botulinum and tetanus neurotoxins (BoNTs and TeNT) are produced as single inactive chains of molecular mass of approximately 150 kDa. Most of these neurotoxins are released after being cleaved into two chains, a heavy chain (HI) of 100 kDa and a light chain (L) of 50 kDa held together by an interchain disulfide bond, by tissue proteinases. BoNT/E is released as a single chain but cleaved by host proteinases [1]. Clostvidium botulinum neurotoxins are extremely poisonous proteins with their LD{sub 50} for humans in the range of 0.1 - 1 ng kg{sup -1} [2]. Botulinum neurotoxins are responsible for neuroparalytic syndromes of botulism characterized by serious neurological disorders and flaccid paralysis. BoNTs block the release of acetylcholine at the neuromuscular junction causing flaccid paralysis while TeNT blocks the release of neurotransmitters like glycine and {gamma}-aminobutyric acid (GABA) in the inhibitory interneurons of the spinal cord resulting in spastic paralysis. In spite of different clinical symptoms, their aetiological agents intoxicate neuronal cells in the same way and these toxins have similar structural organization [3].

  18. Basis Tetrapeptides as Potent Intracellular Inhibitors of type A Botulinum Neurotoxin Protease Activity

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    Hale, M.; Swaminathan, S.; Oyler, G.; Ahmed, S. A.

    2011-01-21

    Botulinum neurotoxins (BoNT) are the most potent of all toxins that cause flaccid muscle paralysis leading to death. They are also potential biothreat agents. A systematic investigation of various short peptide inhibitors of the BoNT protease domain with a 17-residue peptide substrate led to arginine-arginine-glycine-cysteine having a basic tetrapeptide structure as the most potent inhibitor. When assayed in the presence of dithiothreitol (DTT), the inhibitory effect was drastically reduced. Replacing the terminal cysteine with one hydrophobic residue eliminated the DTT effect but with two hydrophobic residues made the pentapeptide a poor inhibitor. Replacing the first arginine with cysteine or adding an additional cysteine at the N terminus did not improve inhibition. When assessed using mouse brain lysates, the tetrapeptides also inhibited BoNT/A cleavage of the endogenous SNAP-25. The peptides penetrated the neuronal cell lines, N2A and BE(2)-M17, without adversely affecting metabolic functions as measured by ATP production and P-38 phosphorylation. Biological activity of the peptides persisted within cultured chick motor neurons and rat and mouse cerebellar neurons for more than 40 h and inhibited BoNT/A protease action inside the neurons in a dose- and time-dependent fashion. Our results define a tetrapeptide as the smallest peptide inhibitor in the backdrop of a large substrate protein of 200+ amino acids having multiple interaction regions with its cognate enzyme. The inhibitors should also be valuable candidates for drug development.

  19. In vitro detection and quantification of botulinum neurotoxin type E activity in avian blood

    Science.gov (United States)

    Piazza, Timothy M.; Blehert, David S.; Dunning, F. Mark; Berlowski-Zier, Brenda M.; Zeytin, Fusun N.; Samuel, Michael D.; Tucker, Ward C.

    2011-01-01

    Botulinum neurotoxin serotype E (BoNT/E) outbreaks in the Great Lakes region cause large annual avian mortality events, with an estimated 17,000 bird deaths reported in 2007 alone. During an outbreak investigation, blood collected from bird carcasses is tested for the presence of BoNT/E using the mouse lethality assay. While sensitive, this method is labor-intensive and low throughput and can take up to 7 days to complete. We developed a rapid and sensitive in vitro assay, the BoTest Matrix E assay, that combines immunoprecipitation with high-affinity endopeptidase activity detection by Förster resonance energy transfer (FRET) to rapidly quantify BoNT/E activity in avian blood with detection limits comparable to those of the mouse lethality assay. On the basis of the analysis of archived blood samples (n = 87) collected from bird carcasses during avian mortality investigations, BoTest Matrix E detected picomolar quantities of BoNT/E following a 2-h incubation and femtomolar quantities of BoNT/E following extended incubation (24 h) with 100% diagnostic specificity and 91% diagnostic sensitivity.

  20. Synergistic capture of Clostridium botulinum Type A neurotoxin by scFv antibodies to novel epitopes

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    Gray, Sean A.; Barr, John R.; Kalb, Suzanne R.; Marks, James D.; Baird, Cheryl L.; Cangelosi, Gerard A.; Miller, Keith D.; Feldhaus, Michael J.

    2011-10-01

    A non-immune library of human single chain fragment variable (scFv) antibodies displayed on Saccharomyces cerevisiae was screened for binding to the Clostridium botulinum neurotoxin serotype A binding domain [BoNT/A (Hc)] with the goal of identifying scFv to novel epitopes. To do this, an antibody-mediated labeling strategy was used in which antigen-binding yeast clones were selected after labeling with previously characterized monoclonal antibodies (MAbs) specific to the Hc. Twenty unique scFv clones were isolated that bound Hc. Of these, three also bound to full-length BoNT/A toxin complex with affinities ranging from 5 nM to 170 nM. Epitope binning showed that the three unique clones recognized at least two epitopes that were distinct from one another and from the detection MAbs. After production in E. coli, the scFv were coupled to magnetic particles and tested for their ability to capture BoNT/A holotoxin using an Endopep-MS assay. In this assay, toxin captured by scFv coated magnetic particles was detected by incubation of the complex with a peptide containing a BoNT/A-specific cleavage sequence. Mass spectrometry was used to detect the ratio of intact peptide to cleavage products as evidence for toxin capture. When tested individually, each of the scFv showed a weak positive Endopep-MS result. However, when the particles were coated with all three scFv simultaneously, they exhibited significantly higher Endopep-MS activity, consistent with synergistic binding. These results demonstrate novel approaches toward the isolation and characterization of scFv antibodies specific to unlabeled antigen. They also provide evidence that distinct scFv antibodies can work synergistically to increase the efficiency of antigen capture onto a solid support.

  1. CRISPR/Cas9-Mediated Genomic Deletion of the Beta-1, 4 N-acetylgalactosaminyltransferase 1 Gene in Murine P19 Embryonal Carcinoma Cells Results in Low Sensitivity to Botulinum Neurotoxin Type C.

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    Kentaro Tsukamoto

    Full Text Available Botulinum neurotoxins produced by Clostridium botulinum cause flaccid paralysis by inhibiting neurotransmitter release at peripheral nerve terminals. Previously, we found that neurons derived from the murine P19 embryonal carcinoma cell line exhibited high sensitivity to botulinum neurotoxin type C. In order to prove the utility of P19 cells for the study of the intracellular mechanism of botulinum neurotoxins, ganglioside-knockout neurons were generated by deletion of the gene encoding beta-1,4 N-acetylgalactosaminyltransferase 1 in P19 cells using the clustered regularly interspaced short palindromic repeats combined with Cas9 (CRISPR/Cas9 system. By using this system, knockout cells could be generated more easily than with previous methods. The sensitivity of the generated beta-1,4 N-acetylgalactosaminyltransferase 1-depleted P19 neurons to botulinum neurotoxin type C was decreased considerably, and the exogenous addition of the gangliosides GD1a, GD1b, and GT1b restored the susceptibility of P19 cells to botulinum neurotoxin type C. In particular, addition of a mixture of these three ganglioside more effectively recovered the sensitivity of knockout cells compared to independent addition of GD1a, GD1b, or GT1b. Consequently, the genome-edited P19 cells generated by the CRISPR/Cas9 system were useful for identifying and defining the intracellular molecules involved in the toxic action of botulinum neurotoxins.

  2. Crystal Structure of the Botulinum Neurotoxin Type G Binding Domain: Insight into Cell Surface Binding

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    Stenmark, Pål; Dong, Min; Dupuy, Jérôme; Chapman, Edwin R.; Stevens, Raymond C. (Scripps); (UW)

    2011-11-02

    Botulinum neurotoxins (BoNTs) typically bind the neuronal cell surface via dual interactions with both protein receptors and gangliosides. We present here the 1.9-{angstrom} X-ray structure of the BoNT serotype G (BoNT/G) receptor binding domain (residues 868-1297) and a detailed view of protein receptor and ganglioside binding regions. The ganglioside binding motif (SxWY) has a conserved structure compared to the corresponding regions in BoNT serotype A and BoNT serotype B (BoNT/B), but several features of interactions with the hydrophilic face of the ganglioside are absent at the opposite side of the motif in the BoNT/G ganglioside binding cleft. This may significantly reduce the affinity between BoNT/G and gangliosides. BoNT/G and BoNT/B share the protein receptor synaptotagmin (Syt) I/II. The Syt binding site has a conserved hydrophobic plateau located centrally in the proposed protein receptor binding interface (Tyr1189, Phe1202, Ala1204, Pro1205, and Phe1212). Interestingly, only 5 of 14 residues that are important for binding between Syt-II and BoNT/B are conserved in BoNT/G, suggesting that the means by which BoNT/G and BoNT/B bind Syt diverges more than previously appreciated. Indeed, substitution of Syt-II Phe47 and Phe55 with alanine residues had little effect on the binding of BoNT/G, but strongly reduced the binding of BoNT/B. Furthermore, an extended solvent-exposed hydrophobic loop, located between the Syt binding site and the ganglioside binding cleft, may serve as a third membrane association and binding element to contribute to high-affinity binding to the neuronal membrane. While BoNT/G and BoNT/B are homologous to each other and both utilize Syt-I/Syt-II as their protein receptor, the precise means by which these two toxin serotypes bind to Syt appears surprisingly divergent.

  3. Modulation of inflammatory mediators in the trigeminal ganglion by botulinum neurotoxin type A

    DEFF Research Database (Denmark)

    Edvinsson, Jacob; Warfvinge, Karin; Edvinsson, Lars

    2015-01-01

    (synaptic vesicle docking protein) or SV2-A (Botulinum toxin receptor element). RESULTS: We report that CGRP, iNOS, IL-1β, SNAP-25 and SV2-A were observed in fresh TG with a differential distribution. Interestingly, NaCl organ culture of the TG resulted in enhanced expression of CGRP and SNAP-25 in neurons...

  4. Therapeutic applications of botulinum neurotoxins in head and neck disorders

    Directory of Open Access Journals (Sweden)

    Ahmad Alshadwi

    2015-01-01

    Conclusion: Botulinum neurotoxin therapy provides viable alternatives to traditional treatment modalities for some conditions affecting the head and neck region that have neurological components. This therapy can overcome some of the morbidities associated with conventional therapy. More research is needed to determine the ideal doses of botulinum neurotoxin to treat different diseases affecting the head and neck regions.

  5. [Botulinum neurotoxin type A in neurogenic detrusor overactivity: consensus paper of the Working Group Neuro-Urology of the DMGP].

    Science.gov (United States)

    Böthig, R; Kaufmann, A; Bremer, J; Pannek, J; Domurath, B

    2014-04-01

    The use of botulinum neurotoxin (BoNT-A) for suppression of neurogenic detrusor overactivity was first reported in 2000. Since that time, this method has gained widespread use. A number of recommendations and consensus statements have already been published. The current practice-oriented consensus paper takes into account recent developments and the over 10-year experience of most members of the Working Group Neuro-Urology of the German-speaking Medical Society for Paraplegia (DMGP) with a focus on the use of BoNT-A in paraplegic patients and in patients with multiple sclerosis. PMID:24604016

  6. A Label Free Colorimetric Assay for the Detection of Active Botulinum Neurotoxin Type A by SNAP-25 Conjugated Colloidal Gold

    Directory of Open Access Journals (Sweden)

    Christopher Gwenin

    2013-08-01

    Full Text Available Botulinum neurotoxins are one of the most potent toxins known to man. Current methods of detection involve the quantification of the toxin but do not take into account the percentage of the toxin that is active. At present the assay used for monitoring the activity of the toxin is the mouse bioassay, which is lengthy and has ethical issues due to the use of live animals. This report demonstrates a novel assay that utilises the endopeptidase activity of the toxin to detect Botulinum neurotoxin in a pharmaceutical sample. The cleaving of SNAP-25 is monitored via UV-Visible spectroscopy with a limit of detection of 373 fg/mL and has been further developed into a high throughput method using a microplate reader detecting down to 600 fg/mL of active toxin. The results show clear differences between the toxin product and the placebo, which contains the pharmaceutical excipients human serum albumin and lactose, showing that the assay detects the active form of the toxin.

  7. Application of Purified Botulinum Type A Neurotoxin to Treat Experimental Trigeminal Neuropathy in Rats and Patients with Urinary Incontinence and Prostatic Hyperplasia

    OpenAIRE

    Yoshizo Matsuka; Teruhiko Yokoyama; Yumiko Yamamoto; Tomonori Suzuki; Ni Nengah Dwi Fatmawati; Atsushi Nishikawa; Tohru Ohyama; Toshihiro Watanabe; Takuo Kuboki; Atsushi Nagai; Keiji Oguma

    2012-01-01

    Type A neurotoxin (NTX) of Clostridium botulinum was purified by a simple procedure using a lactose gel column. The toxicity of this purified toxin preparation was retained for at least 1 year at −30°C by supplementation with either 0.1% albumin or 0.05% albumin plus 1% trehalose. When purified NTX was used to treat 49 patients with urinary incontinence caused by either refractory idiopathic or neurogenic detrusor overactivity, 36 patients showed significant improvement in symptoms. These ben...

  8. Universal and specific quantitative detection of botulinum neurotoxin genes

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    Arnon Stephen S

    2010-10-01

    Full Text Available Abstract Background 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-accepted assay to detect and type botulinum neurotoxins (BoNTs is the mouse protection bioassay. While specific and sensitive, this assay requires the use of laboratory animals, may take up to four days to achieve a diagnosis, and is unsuitable for high-throughput analysis. We report here a two-step PCR assay that identifies all toxin types, that achieves the specificity of the mouse bioassay while surpassing it in equivalent sensitivity, that has capability for high-throughput analysis, and that provides quantitative results within hours. The first step of our assay consists of a conventional PCR that detects the presence of C. botulinum regardless of the neurotoxin type. The second step uses quantitative PCR (qPCR technology to determine the specific serotype of the neurotoxin. Results We assayed purified C. botulinum DNA and crude toxin preparations, as well as food and stool from healthy individuals spiked with purified BoNT DNA, and one stool sample from a case of infant botulism for the presence of the NTNH gene, which is part of the BoNT gene cluster, and for the presence of serotype-specific BoNT genes. The PCR surpassed the mouse bioassay both in specificity and sensitivity, detecting positive signals in BoNT preparations containing well below the 1 LD50 required for detection via the mouse bioassay. These results were type-specific and we were reliably able to quantify as few as 10 genomic copies. Conclusions While other studies

  9. Development of a Highly Sensitive Cell-Based Assay for Detecting Botulinum Neurotoxin Type A through Neural Culture Media Optimization.

    Science.gov (United States)

    Hong, Won S; Pezzi, Hannah M; Schuster, Andrea R; Berry, Scott M; Sung, Kyung E; Beebe, David J

    2016-01-01

    Botulinum neurotoxin (BoNT) is the most lethal naturally produced neurotoxin. Due to the extreme toxicity, BoNTs are implicated in bioterrorism, while the specific mechanism of action and long-lasting effect was found to be medically applicable in treating various neurological disorders. Therefore, for both public and patient safety, a highly sensitive, physiologic, and specific assay is needed. In this paper, we show a method for achieving a highly sensitive cell-based assay for BoNT/A detection using the motor neuron-like continuous cell line NG108-15. To achieve high sensitivity, we performed a media optimization study evaluating three commercially available neural supplements in combination with retinoic acid, purmorphamine, transforming growth factor β1 (TGFβ1), and ganglioside GT1b. We found nonlinear combinatorial effects on BoNT/A detection sensitivity, achieving an EC50 of 7.4 U ± 1.5 SD (or ~7.9 pM). The achieved detection sensitivity is comparable to that of assays that used primary and stem cell-derived neurons as well as the mouse lethality assay.

  10. Development of recombinant vaccines for botulinum neurotoxin.

    Science.gov (United States)

    Smith, L A

    1998-11-01

    Synthetic genes encoding non-toxic, carboxyl-terminal regions (approximately 50 kDa) of botulinum neurotoxin (BoNT) serotypes A and B (referred to as fragment C or HC) were constructed and cloned into the methylotropic yeast, Pichia pastoris. Genes specifying BoNTA(HC) and BoNTB(HC) were expressed as both intracellular and secreted products. Recombinants, expressed intracellularly, yielded products with the expected molecular weight as judged by SDS PAGE and Western blot (immunoblot) analysis, while secreted products were larger due to glycosylation. Gene products were used to vaccinate mice and evaluated for their ability to elicit protective antibody titers in vivo. Mice given three intramuscular vaccinations with yeast supernatant containing glycosylated BoNTA(HC) were protected against an intraperitoneal challenge of 10(6) 50% mouse lethal doses (MLD50) of serotype A neurotoxin, a result not duplicated by its BoNTB(HC) counterpart. Vaccinating mice with cytoplasmically produced BoNTA(HC) and BoNTB(HC) protected animals from a challenge of 10(6) MLD50 of serotype A and B toxins, respectively. Because of the glycosylation encountered with secreted BoNT(HC), our efforts focused on the production and purification of products from intracellular expression. PMID:9792170

  11. Structure of botulinum neurotoxin type D light chain at 1.65 A resolution: repercussions for VAMP-2 substrate specificity.

    Science.gov (United States)

    Arndt, Joseph W; Chai, Qing; Christian, Todd; Stevens, Raymond C

    2006-03-14

    The seven serotypes (A-G) of botulinum neurotoxins (BoNTs) function through their proteolytic cleavage of one of three proteins (SNAP-25, Syntaxin, and VAMP) that form the SNARE complex required for synaptic vesicle fusion. The different BoNTs have very specific protease recognition requirements, between 15 and 50 amino acids in length depending on the serotype. However, the structural details involved in substrate recognition remain largely unknown. Here is reported the 1.65 A resolution crystal structure of the catalytic domain of BoNT serotype D (BoNT/D-LC), providing insight into the protein-protein binding interaction and final proteolysis of VAMP-2. Structural analysis has identified a hydrophobic pocket potentially involved in substrate recognition of the P1' VAMP residue (Leu 60) and a second remote site for recognition of the V1 SNARE motif that is critical for activity. A structural comparison of BoNT/D-LC with BoNT/F-LC that also recognizes VAMP-2 one residue away from the BoNT/D-LC site provides additional molecular details about the unique serotype specific activities. In particular, BoNT/D prefers a hydrophobic interaction for the V1 motif of VAMP-2, while BoNT/F adopts a more hydrophilic strategy for recognition of the same V1 motif.

  12. Functional influence of botulinum neurotoxin type A treatment (Xeomin® of multifocal upper and lower limb spasticity on chronic hemiparetic gait

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    Maurizio Falso

    2012-05-01

    Full Text Available This report describes the modification of hemiplegic shoulder pain and walking velocity through injections of Xeomin®, a new botulinum neurotoxin type A formulation, in a 67-year-old woman with chronic residual left hemiparesis and hemiparetic gait attributable to stroke. Clinical evaluation included upper and lower limb spasticity, upper and lower limb pain, trunk control, upper and lower limb motricity index, visual gait analysis, and gait velocity. Assessments were performed before, 1 week after, and 1 month after treatment. Improvement was observed in all clinical parameters assessed. Amelioration of spasticity of the upper and lower limbs and shoulder pain was observed after 1 month. Trunk postural attitude and paraxial muscle recruitment recovered. No adverse events were observed and the patient shows significant improvement of functional impairment derived from chronic spasticity after treatment with Xeomin®. We also provide a simple and useful protocol for clinical evaluation of the treatment.

  13. Crystal Structure of Botulinum Neurotoxin Type a in Complex With the Cell Surface Co-Receptor GT1b-Insight Into the Toxin-Neuron Interaction

    Energy Technology Data Exchange (ETDEWEB)

    Stenmark, P.; Dupuy, J.; Inamura, A.; Kiso, M.; Stevens, R.C.

    2009-05-26

    Botulinum neurotoxins have a very high affinity and specificity for their target cells requiring two different co-receptors located on the neuronal cell surface. Different toxin serotypes have different protein receptors; yet, most share a common ganglioside co-receptor, GT1b. We determined the crystal structure of the botulinum neurotoxin serotype A binding domain (residues 873-1297) alone and in complex with a GT1b analog at 1.7 A and 1.6 A, respectively. The ganglioside GT1b forms several key hydrogen bonds to conserved residues and binds in a shallow groove lined by Tryptophan 1266. GT1b binding does not induce any large structural changes in the toxin; therefore, it is unlikely that allosteric effects play a major role in the dual receptor recognition. Together with the previously published structures of botulinum neurotoxin serotype B in complex with its protein co-receptor, we can now generate a detailed model of botulinum neurotoxin's interaction with the neuronal cell surface. The two branches of the GT1b polysaccharide, together with the protein receptor site, impose strict geometric constraints on the mode of interaction with the membrane surface and strongly support a model where one end of the 100 A long translocation domain helix bundle swing into contact with the membrane, initiating the membrane anchoring event.

  14. In vitro peptide cleavage assay for detection of Botulinum Neurotoxin-A activity in food

    Science.gov (United States)

    The gold standard assay for measuring the activity and typing of Clostridium botulinum neurotoxins is the mouse bioassay. The mouse bioassay is sensitive, robust and does not require specialized equipment. However, the mouse bioassay is slow, not practical for many settings and results in the death ...

  15. Independent evolution of neurotoxin and flagellar genetic loci in proteolytic Clostridium botulinum

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    Twine Susan M

    2009-03-01

    Full Text Available Abstract Background Proteolytic Clostridium botulinum is the causative agent of botulism, a severe neuroparalytic illness. Given the severity of botulism, surprisingly little is known of the population structure, biology, phylogeny or evolution of C. botulinum. The recent determination of the genome sequence of C. botulinum has allowed comparative genomic indexing using a DNA microarray. Results Whole genome microarray analysis revealed that 63% of the coding sequences (CDSs present in reference strain ATCC 3502 were common to all 61 widely-representative strains of proteolytic C. botulinum and the closely related C. sporogenes tested. This indicates a relatively stable genome. There was, however, evidence for recombination and genetic exchange, in particular within the neurotoxin gene and cluster (including transfer of neurotoxin genes to C. sporogenes, and the flagellar glycosylation island (FGI. These two loci appear to have evolved independently from each other, and from the remainder of the genetic complement. A number of strains were atypical; for example, while 10 out of 14 strains that formed type A1 toxin gave almost identical profiles in whole genome, neurotoxin cluster and FGI analyses, the other four strains showed divergent properties. Furthermore, a new neurotoxin sub-type (A5 has been discovered in strains from heroin-associated wound botulism cases. For the first time, differences in glycosylation profiles of the flagella could be linked to differences in the gene content of the FGI. Conclusion Proteolytic C. botulinum has a stable genome backbone containing specific regions of genetic heterogeneity. These include the neurotoxin gene cluster and the FGI, each having evolved independently of each other and the remainder of the genetic complement. Analysis of these genetic components provides a high degree of discrimination of strains of proteolytic C. botulinum, and is suitable for clinical and forensic investigations of botulism

  16. Botulinum Neurotoxin Serotypes Detected by Electrochemical Impedance Spectroscopy

    Science.gov (United States)

    Savage, Alison C.; Buckley, Nicholas; Halliwell, Jennifer; Gwenin, Christopher

    2015-01-01

    Botulinum neurotoxin is one of the deadliest biological toxins known to mankind and is able to cause the debilitating disease botulism. The rapid detection of the different serotypes of botulinum neurotoxin is essential for both diagnosis of botulism and identifying the presence of toxin in potential cases of terrorism and food contamination. The modes of action of botulinum neurotoxins are well-established in literature and differ for each serotype. The toxins are known to specifically cleave portions of the SNARE proteins SNAP-25 or VAMP; an interaction that can be monitored by electrochemical impedance spectroscopy. This study presents a SNAP-25 and a VAMP biosensors for detecting the activity of five botulinum neurotoxin serotypes (A–E) using electrochemical impedance spectroscopy. The biosensors are able to detect concentrations of toxins as low as 25 fg/mL, in a short time-frame compared with the current standard methods of detection. Both biosensors show greater specificity for their compatible serotypes compared with incompatible serotypes and denatured toxins. PMID:25954998

  17. Botulinum Neurotoxin Serotypes Detected by Electrochemical Impedance Spectroscopy

    Directory of Open Access Journals (Sweden)

    Alison C. Savage

    2015-05-01

    Full Text Available Botulinum neurotoxin is one of the deadliest biological toxins known to mankind and is able to cause the debilitating disease botulism. The rapid detection of the different serotypes of botulinum neurotoxin is essential for both diagnosis of botulism and identifying the presence of toxin in potential cases of terrorism and food contamination. The modes of action of botulinum neurotoxins are well-established in literature and differ for each serotype. The toxins are known to specifically cleave portions of the SNARE proteins SNAP-25 or VAMP; an interaction that can be monitored by electrochemical impedance spectroscopy. This study presents a SNAP-25 and a VAMP biosensors for detecting the activity of five botulinum neurotoxin serotypes (A–E using electrochemical impedance spectroscopy. The biosensors are able to detect concentrations of toxins as low as 25 fg/mL, in a short time-frame compared with the current standard methods of detection. Both biosensors show greater specificity for their compatible serotypes compared with incompatible serotypes and denatured toxins.

  18. Identification of the amino acid residues rendering TI-VAMP insensitive toward botulinum neurotoxin B.

    Science.gov (United States)

    Sikorra, Stefan; Henke, Tina; Swaminathan, Subramanyam; Galli, Thierry; Binz, Thomas

    2006-03-24

    Botulinum neurotoxins types B, D, F, and G, and tetanus neurotoxin inhibit vesicular fusion via proteolytic cleavage of VAMP/Synaptobrevin, a core component of the membrane fusion machinery. Thus, these neurotoxins became widely used tools for investigating vesicular trafficking routes. Except for VAMP-1, VAMP-2, and Cellubrevin, no other member of the VAMP family represents a substrate for these neurotoxins. The molecular basis for this discrepancy is not known. A 34 amino acid residue segment of VAMP-2 was previously suggested to mediate the interaction with botulinum neurotoxin B, but the validity of the data was later questioned. To check whether this segment alone controls the susceptibility toward botulinum neurotoxin B, it was used to replace the corresponding segment in TI-VAMP. The resulting VAMP hybrid and VAMP-2 were hydrolysed at virtually identical rates. Resetting the VAMP-2 portion in the hybrid from either end to TI-VAMP residues gradually reduced the cleavability. A hybrid encompassing merely the VAMP-2 segment 71-80 around the Gln76/Phe77 scissile bond was still hydrolysed, albeit at a approximately tenfold lower cleavage rate. The contribution of each non-conserved amino acid of the whole 34-mer segment to the interaction was investigated employing VAMP-2. We find that the eight non-conserved residues of the 71-80 segment are all necessary for efficient cleavage. Mutation of an additional six residues located upstream and downstream of this segment affects substrate hydrolysis as well. Vice versa, a readily cleavable TI-VAMP molecule requires at the least the replacement of Ile158, Thr161, and the section 165-174 by Asp64, Ala67, and the 71-80 segment of VAMP-2, respectively. However, the insensitivity of TI-VAMP to botulinum neurotoxin B relies on at least 12 amino acid changes versus VAMP-2. These are scattered along an interface of 22 amino acid residues in length.

  19. Application of Purified Botulinum Type A Neurotoxin to Treat Experimental Trigeminal Neuropathy in Rats and Patients with Urinary Incontinence and Prostatic Hyperplasia

    Directory of Open Access Journals (Sweden)

    Yoshizo Matsuka

    2012-01-01

    Full Text Available Type A neurotoxin (NTX of Clostridium botulinum was purified by a simple procedure using a lactose gel column. The toxicity of this purified toxin preparation was retained for at least 1 year at −30°C by supplementation with either 0.1% albumin or 0.05% albumin plus 1% trehalose. When purified NTX was used to treat 49 patients with urinary incontinence caused by either refractory idiopathic or neurogenic detrusor overactivity, 36 patients showed significant improvement in symptoms. These beneficial effects were also observed in cases of prostatic hyperplasia. The results obtained with NTX were similar to that of Botox. The effects of NTX on trigeminal neuralgia induced by infraorbital nerve constriction (IoNC in rats were also studied. Trigeminal ganglion neurons from ipsilateral to IoNC exhibited significantly faster onset of FM4-64 release than sham-operated contralateral neurons. Intradermal injection of NTX in the area of IoNC alleviated IoNC-induced pain behavior and reduced the exaggerated FM4-64 release in trigeminal ganglion neurons.

  20. Application of purified botulinum type a neurotoxin to treat experimental trigeminal neuropathy in rats and patients with urinary incontinence and prostatic hyperplasia.

    Science.gov (United States)

    Matsuka, Yoshizo; Yokoyama, Teruhiko; Yamamoto, Yumiko; Suzuki, Tomonori; Dwi Fatmawati, Ni Nengah; Nishikawa, Atsushi; Ohyama, Tohru; Watanabe, Toshihiro; Kuboki, Takuo; Nagai, Atsushi; Oguma, Keiji

    2012-01-01

    Type A neurotoxin (NTX) of Clostridium botulinum was purified by a simple procedure using a lactose gel column. The toxicity of this purified toxin preparation was retained for at least 1 year at -30°C by supplementation with either 0.1% albumin or 0.05% albumin plus 1% trehalose. When purified NTX was used to treat 49 patients with urinary incontinence caused by either refractory idiopathic or neurogenic detrusor overactivity, 36 patients showed significant improvement in symptoms. These beneficial effects were also observed in cases of prostatic hyperplasia. The results obtained with NTX were similar to that of Botox. The effects of NTX on trigeminal neuralgia induced by infraorbital nerve constriction (IoNC) in rats were also studied. Trigeminal ganglion neurons from ipsilateral to IoNC exhibited significantly faster onset of FM4-64 release than sham-operated contralateral neurons. Intradermal injection of NTX in the area of IoNC alleviated IoNC-induced pain behavior and reduced the exaggerated FM4-64 release in trigeminal ganglion neurons. PMID:22745637

  1. The effect of pH on recombinant C-terminal domain of Botulinum Neurotoxin type E (rBoNT/E-HCC

    Directory of Open Access Journals (Sweden)

    Seyed Jafar Mousavy

    2013-12-01

    Full Text Available Recombinant proteins are tending to be the most favorable vaccine-candidates against botulism. Recombinant Carboxy-terminal of botulinum neurotoxin serotype E (rBoNT/E-HCC has been introduced as an efficient vaccine against botulism type E. In this report, we made an effort to investigate the effect of different pH on protein structure to assess if rBoNT/E-HCC could be used as a vaccine for oral administration. Initially, rBoNT/E-HCC was expressed and purified. Structural changes of rBoNT/E-HCC at several pH conditions were studied by various techniques including circular dichroism (CD, fluorescence, aggregation and UV-Vis spectroscopy. The results showed the more compact and more stable structure for rBoNT/E-HCC at acidic pH, and loosely folded structure at alkaline pH. Our finding as the first step of rBoNT/E-HCC evaluation, hopefully introduce it as a suitable vaccine candidate for oral administration.

  2. Structure- and Substrate- Based Inhibitor Design for Clostridium botulinum Neurotoxin Serotype A*

    Energy Technology Data Exchange (ETDEWEB)

    Kumaran,D.; Rawat, R.; Ludivico, M.; Ahmed, S.; Swaminathan, S.

    2008-01-01

    The seven antigenically distinct serotypes of Clostridium botulinum neurotoxins cleave specific soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex proteins and block the release of neurotransmitters that cause flaccid paralysis and are considered potential bioweapons. Botulinum neurotoxin type A is the most potent among the clostridial neurotoxins, and to date there is no post-exposure therapeutic intervention available. To develop inhibitors leading to drug design, it is imperative that critical interactions between the enzyme and the substrate near the active site are known. Although enzyme-substrate interactions at exosites away from the active site are mapped in detail for botulinum neurotoxin type A, information about the active site interactions is lacking. Here, we present the crystal structures of botulinum neurotoxin type A catalytic domain in complex with four inhibitory substrate analog tetrapeptides, viz. RRGC, RRGL, RRGI, and RRGM at resolutions of 1.6-1.8 Angstroms . These structures show for the first time the interactions between the substrate and enzyme at the active site and delineate residues important for substrate stabilization and catalytic activity. We show that OH of Tyr366 and NH2 of Arg363 are hydrogen-bonded to carbonyl oxygens of P1 and P1' of the substrate analog and position it for catalytic activity. Most importantly, the nucleophilic water is replaced by the amino group of the N-terminal residue of the tetrapeptide. Furthermore, the S1' site is formed by Phe194, Thr215, Thr220, Asp370, and Arg363. The Ki of the best inhibitory tetrapeptide is 157 nm.

  3. Electrochemical impedance spectroscopy biosensor for detection of active botulinum neurotoxin

    Directory of Open Access Journals (Sweden)

    Jennifer Halliwell

    2014-12-01

    Full Text Available The standard method for the detection of botulinum neurotoxin is currently the mouse bioassay which is considered to be the most reliable method for the detection of the active form of this toxin. Despite this it is a time-consuming and expensive assay to run and as such many alternative assays have recently been proposed. Herein we report the development of two electrochemical assays for the detection of active botulinum neurotoxin in a pharmaceutical sample. Gold electrodes were modified with self-assembled monolayers of the SNARE protein SNAP-25 which is selectively cleaved by active botulinum neurotoxin A. Cyclic voltammetry and electrochemical impedance spectroscopy were performed on the modified working electrodes to observe changes to the layer on addition of the toxin. Both methods were able to distinguish the difference between the presence of the active toxin and a placebo containing the excipients of the pharmaceutical product. The electrochemical impedance spectroscopy assay also allowed for detection of the active toxin at concentrations as low as 25 fg/ml, with results being obtained in under an hour outperforming the mouse bioassay.

  4. Time Course Analysis of the Effects of Botulinum Neurotoxin Type A on Pain and Vasomotor Responses Evoked by Glutamate Injection into Human Temporalis Muscles

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    Larissa Bittencourt da Silva

    2014-02-01

    Full Text Available The effect of botulinum neurotoxin type A (BoNTA on glutamate-evoked temporalis muscle pain and vasomotor responses was investigated in healthy men and women over a 60 day time course. Subjects participated in a pre-BoNTA session where their responses to injection of glutamate (1 M, 0.2 mL and saline (0.2 mL into the temporalis muscles were assessed. On Day 1, BoNTA (5 U was injected into one temporalis muscle and saline into the contralateral temporalis muscle, in a randomized order. Subjects then received intramuscular injections of glutamate (1 M, 0.2 mL into the left and right temporalis muscles at 3 h and subsequently 7, 30 and 60 days post-injection of BoNTA. Pain intensity, pain area, and neurogenic inflammation (skin temperature and skin blood perfusion were recorded. Prior to BoNTA treatment, glutamate evoked significantly greater pain and vasomotor reactions (P < 0.001 than saline. BoNTA significantly reduced glutamate-evoked pain intensity (P < 0.05, pain area (P < 0.01, skin blood perfusion (P < 0.05, and skin temperature (P < 0.001. The inhibitory effect of BoNTA was present at 3 h after injection, peaked after 7 days and returned to baseline by 60 days. Findings from the present study demonstrated a rapid action of BoNTA on glutamate-evoked pain and neurogenic inflammation, which is in line with animal studies.

  5. Characterization of botulinum neurotoxin type A neutralizing monoclonal antibodies and influence of their half-lives on therapeutic activity.

    Directory of Open Access Journals (Sweden)

    Christelle Mazuet

    Full Text Available Botulinum toxins, i.e. BoNT/A to/G, include the most toxic substances known. Since botulism is a potentially fatal neuroparalytic disease with possible use as a biowarfare weapon (Centers for Disease Control and Prevention category A bioterrorism agent, intensive efforts are being made to develop vaccines or neutralizing antibodies. The use of active fragments from non-human immunoglobulins (F(ab'(2, Fab', scFv, chemically modified or not, may avoid side effects, but also largely modify the in vivo half-life and effectiveness of these reagents. We evaluated the neutralizing activity of several monoclonal anti-BoNT/A antibodies (mAbs. F(ab'(2 fragments, native or treated with polyethyleneglycol (PEG, were prepared from selected mAbs to determine their half-life and neutralizing activity as compared with the initial mAbs. We compared the protective efficiency of the different biochemical forms of anti-toxin mAbs providing the same neutralizing activity. Among fourteen tested mAbs, twelve exhibited neutralizing activity. Fragments from two of the best mAbs (TA12 and TA17, recognizing different epitopes, were produced. These two mAbs neutralized the A1 subtype of the toxin more efficiently than the A2 or A3 subtypes. Since mAb TA12 and its fragments both exhibited the greatest neutralizing activity, they were further evaluated in the therapeutic experiments. These showed that, in a mouse model, a 2- to 4-h interval between toxin and antitoxin injection allows the treatment to remain effective, but also suggested an absence of correlation between the half-life of the antitoxins and the length of time before treatment after botulinum toxin A contamination. These experiments demonstrate that PEG treatment has a strong impact on the half-life of the fragments, without affecting the effectiveness of neutralization, which was maintained after preparation of the fragments. These reagents may be useful for rapid treatment after botulinum toxin A

  6. Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E

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    Uma Basavanna

    2013-01-01

    Full Text Available The standard procedure for definitive detection of BoNT-producing Clostridia is a culture method combined with neurotoxin detection using a standard mouse bioassay (MBA. The mouse bioassay is highly sensitive and specific, but it is expensive and time-consuming, and there are ethical concerns due to use of laboratory animals. Cell-based assays provide an alternative to the MBA in screening for BoNT-producing Clostridia. Here, we describe a cell-based assay utilizing a fluorescence reporter construct expressed in a neuronal cell model to study toxin activity in situ. Our data indicates that the assay can detect as little as 100 pM BoNT/A activity within living cells, and the assay is currently being evaluated for the analysis of BoNT in food matrices. Among available in vitro assays, we believe that cell-based assays are widely applicable in high-throughput screenings and have the potential to at least reduce and refine animal assays if not replace it.

  7. Botulinum Neurotoxin Is Shielded by NTNHA in an Interlocked Complex

    Energy Technology Data Exchange (ETDEWEB)

    Gu, Shenyan; Rumpel, Sophie; Zhou, Jie; Strotmeier, Jasmin; Bigalke, Hans; Perry, Kay; Shoemaker, Charles B.; Rummel, Andreas; Jin, Rongsheng (Cornell); (Tufts); (Hannover-MED); (Sanford-Burnham)

    2012-03-28

    Botulinum neurotoxins (BoNTs) are highly poisonous substances that are also effective medicines. Accidental BoNT poisoning often occurs through ingestion of Clostridium botulinum-contaminated food. Here, we present the crystal structure of a BoNT in complex with a clostridial nontoxic nonhemagglutinin (NTNHA) protein at 2.7 angstroms. Biochemical and functional studies show that NTNHA provides large and multivalent binding interfaces to protect BoNT from gastrointestinal degradation. Moreover, the structure highlights key residues in BoNT that regulate complex assembly in a pH-dependent manner. Collectively, our findings define the molecular mechanisms by which NTNHA shields BoNT in the hostile gastrointestinal environment and releases it upon entry into the circulation. These results will assist in the design of small molecules for inhibiting oral BoNT intoxication and of delivery vehicles for oral administration of biologics.

  8. Genetic Diversity Among Botulinum Neurotoxin Producing Clostridial Strains

    Energy Technology Data Exchange (ETDEWEB)

    Hill, K K; Smith, T J; Helma, C H; Ticknor, L O; Foley, B T; Svennson, R T; Brown, J L; Johnson, E A; Smith, L A; Okinaka, R T; Jackson, P J; Marks, J D

    2006-07-06

    Clostridium botulinum is a taxonomic designation for many diverse anaerobic spore forming rod-shaped bacteria which have the common property of producing botulinum neurotoxins (BoNTs). The BoNTs are exoneurotoxins that can cause severe paralysis and even death in humans and various other animal species. A collection of 174 C. botulinum strains were examined by amplified fragment length polymorphism (AFLP) analysis and by sequencing of the 16S rRNA gene and BoNT genes to examine genetic diversity within this species. This collection contained representatives of each of the seven different serotypes of botulinum neurotoxins (BoNT A-G). Analysis of the16S rRNA sequences confirmed earlier reports of at least four distinct genomic backgrounds (Groups I-IV) each of which has independently acquired one or more BoNT serotypes through horizontal gene transfer. AFLP analysis provided higher resolution, and can be used to further subdivide the four groups into sub-groups. Sequencing of the BoNT genes from serotypes A, B and E in multiple strains confirmed significant sequence variation within each serotype. Four distinct lineages within each of the BoNT A and B serotypes, and five distinct lineages of serotype E strains were identified. The nucleotide sequences of the seven serotypes of BoNT were compared and show varying degrees of interrelatedness and recombination as has been previously noted for the NTNH gene which is linked to BoNT. These analyses contribute to the understanding of the evolution and phylogeny within this species and assist in the development of improved diagnostics and therapeutics for treatment of botulism.

  9. Treating benign prostatic hyperplasia with botulinum neurotoxin.

    Science.gov (United States)

    Brisinda, G; Vanella, S; Marniga, G; Crocco, A; Maria, G

    2011-01-01

    Botulinum toxin (BoNT) has been increasingly used in the interventional treatment of several disorders; the use of this agent has extended to a plethora of conditions including focal dystonia, spasticity, inappropriate contraction in most gastrointestinal sphincters, eye movement disorders, hyperhidrosis, genitourinary disorders and aesthetically undesirable hyperfunctional facial lines. In addition, BoNT is being investigated for the control of pain, and for the management of tension or migraine headaches and myofascial pain syndrome. Benign prostatic hyperplasia (BPH) is a common condition in ageing men; the goal of therapy is to reduce the lower urinary tract symptoms (LUTS) associated with BPH and to improve the quality of life. However, medical treatment, including drugs that relax smooth muscle within the prostate and drugs that shrink the gland are not totally effective or without complications. The standard surgical treatment for BPH is progressively changing to minimally invasive therapies, but none of them has provided clear results. The use of BoNT-A to inhibit the autonomic efferent effects on prostate growth and contraction, and inhibit the abnormal afferent effects on prostate sensation, might be an alternative treatment for BPH. BoNT injections have several advantages over drugs and surgical therapies in the management of intractable or chronic disease; systemic pharmacologic effects are rare, permanent destruction of tissue does not occur, and graded degrees of relaxation may be achieved by varying the dose injected. In this paper, clinical experience over the last years with BoNT in BPH impaired patients will be illustrated.

  10. Expression and Purification of Neurotoxin-Associated Protein HA-33/A from Clostridium botulinum and Evaluation of Its Antigenicity

    OpenAIRE

    Sayadmanesh, Ali; Ebrahimi, Firouz; Hajizade, Abbas; Rostamian, Mosayeb; Keshavarz, Hani

    2013-01-01

    Background: Botulinum neurotoxin (BoNT) complexes consist of neurotoxin and neurotoxin-associated proteins. Hemagglutinin-33 (HA-33) is a member of BoNT type A (BoNT/A) complex. Considering the protective role of HA-33 in preservation of BoNT/A in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. Thus in this study, HA-33 was expressed and purified, and subsequently its antigenicity in mice was studied. Methods: Initially, ha-33...

  11. Substrate binding mode and its implication on drug design for botulinum neurotoxin A.

    Directory of Open Access Journals (Sweden)

    Desigan Kumaran

    Full Text Available The seven antigenically distinct serotypes of Clostridium botulinum neurotoxins, the causative agents of botulism, block the neurotransmitter release by specifically cleaving one of the three SNARE proteins and induce flaccid paralysis. The Centers for Disease Control and Prevention (CDC has declared them as Category A biowarfare agents. The most potent among them, botulinum neurotoxin type A (BoNT/A, cleaves its substrate synaptosome-associated protein of 25 kDa (SNAP-25. An efficient drug for botulism can be developed only with the knowledge of interactions between the substrate and enzyme at the active site. Here, we report the crystal structures of the catalytic domain of BoNT/A with its uncleavable SNAP-25 peptide (197QRATKM(202 and its variant (197RRATKM(202 to 1.5 A and 1.6 A, respectively. This is the first time the structure of an uncleavable substrate bound to an active botulinum neurotoxin is reported and it has helped in unequivocally defining S1 to S5' sites. These substrate peptides make interactions with the enzyme predominantly by the residues from 160, 200, 250 and 370 loops. Most notably, the amino nitrogen and carbonyl oxygen of P1 residue (Gln197 chelate the zinc ion and replace the nucleophilic water. The P1'-Arg198, occupies the S1' site formed by Arg363, Thr220, Asp370, Thr215, Ile161, Phe163 and Phe194. The S2' subsite is formed by Arg363, Asn368 and Asp370, while S3' subsite is formed by Tyr251, Leu256, Val258, Tyr366, Phe369 and Asn388. P4'-Lys201 makes hydrogen bond with Gln162. P5'-Met202 binds in the hydrophobic pocket formed by the residues from the 250 and 200 loop. Knowledge of interactions between the enzyme and substrate peptide from these complex structures should form the basis for design of potent inhibitors for this neurotoxin.

  12. Purification and Characterization of Botulinum Neurotoxin FA from a Genetically Modified Clostridium botulinum Strain.

    Science.gov (United States)

    Pellett, Sabine; Tepp, William H; Bradshaw, Marite; Kalb, Suzanne R; Dykes, Janet K; Lin, Guangyun; Nawrocki, Erin M; Pier, Christina L; Barr, John R; Maslanka, Susan E; Johnson, Eric A

    2016-01-01

    Botulinum neurotoxins (BoNTs), produced by neurotoxigenic clostridial species, are the cause of the severe disease botulism in humans and animals. Early research on BoNTs has led to their classification into seven serotypes (serotypes A to G) based upon the selective neutralization of their toxicity in mice by homologous antibodies. Recently, a report of a potential eighth serotype of BoNT, designated "type H," has been controversial. This novel BoNT was produced together with BoNT/B2 in a dual-toxin-producing Clostridium botulinum strain. The data used to designate this novel toxin as a new serotype were derived from culture supernatant containing both BoNT/B2 and novel toxin and from sequence information, although data from two independent laboratories indicated neutralization by antibodies raised against BoNT/A1, and classification as BoNT/FA was proposed. The sequence data indicate a chimeric structure consisting of a BoNT/A1 receptor binding domain, a BoNT/F5 light-chain domain, and a novel translocation domain most closely related to BoNT/F1. Here, we describe characterization of this toxin purified from the native strain in which expression of the second BoNT (BoNT/B) has been eliminated. Mass spectrometry analysis indicated that the toxin preparation contained only BoNT/FA and confirmed catalytic activity analogous to that of BoNT/F5. The in vivo mouse bioassay indicated a specific activity of this toxin of 3.8 × 10(7) mouse 50% lethal dose (mLD50) units/mg, whereas activity in cultured human neurons was very high (50% effective concentration [EC50] = 0.02 mLD50/well). Neutralization assays in cells and mice both indicated full neutralization by various antibodies raised against BoNT/A1, although at 16- to 20-fold-lower efficiency than for BoNT/A1. IMPORTANCE Botulinum neurotoxins (BoNTs), produced by anaerobic bacteria, are the cause of the potentially deadly, neuroparalytic disease botulism. BoNTs have been classified into seven serotypes, serotypes A

  13. Substrate Recognition of VAMP-2 by Botulinum Neurotoxin B and Tetanus Neurotoxin*

    Science.gov (United States)

    Chen, Sheng; Hall, Cherisse; Barbieri, Joseph T.

    2008-01-01

    Botulinum neurotoxin (BoNT; serotypes A-G) and tetanus neurotoxin elicit flaccid and spastic paralysis, respectively. These neurotoxins are zinc proteases that cleave SNARE proteins to inhibit synaptic vesicle fusion to the plasma membrane. Although BoNT/B and tetanus neurotoxin (TeNT) cleave VAMP-2 at the same scissile bond, their mechanism(s) of VAMP-2 recognition is not clear. Mapping experiments showed that residues 60-87 of VAMP-2 were sufficient for efficient cleavage by BoNT/B and that residues 40-87 of VAMP-2 were sufficient for efficient TeNT cleavage. Alanine-scanning mutagenesis and kinetic analysis identified three regions within VAMP-2 that were recognized by BoNT/B and TeNT: residues adjacent to the site of scissile bond cleavage (cleavage region) and residues located within N-terminal and C-terminal regions relative to the cleavage region. Analysis of residues within the cleavage region showed that mutations at the P7, P4, P2, and P1′ residues of VAMP-2 had the greatest inhibition of LC/B cleavage (≥32- fold), whereas mutations at P7, P4, P1′, and P2′ residues of VAMP-2 had the greatest inhibition of LC/TeNT cleavage (≥64-fold). Residues within the cleavage region influenced catalysis, whereas residues N-terminal and C-terminal to the cleavage region influenced binding affinity. Thus, BoNT/B and TeNT possess similar organization but have unique residues to recognize and cleave VAMP-2. These studies provide new insights into how the clostridial neurotoxins recognize their substrates. PMID:18511417

  14. Substrate recognition of VAMP-2 by botulinum neurotoxin B and tetanus neurotoxin.

    Science.gov (United States)

    Chen, Sheng; Hall, Cherisse; Barbieri, Joseph T

    2008-07-25

    Botulinum neurotoxin (BoNT; serotypes A-G) and tetanus neurotoxin elicit flaccid and spastic paralysis, respectively. These neurotoxins are zinc proteases that cleave SNARE proteins to inhibit synaptic vesicle fusion to the plasma membrane. Although BoNT/B and tetanus neurotoxin (TeNT) cleave VAMP-2 at the same scissile bond, their mechanism(s) of VAMP-2 recognition is not clear. Mapping experiments showed that residues 60-87 of VAMP-2 were sufficient for efficient cleavage by BoNT/B and that residues 40-87 of VAMP-2 were sufficient for efficient TeNT cleavage. Alanine-scanning mutagenesis and kinetic analysis identified three regions within VAMP-2 that were recognized by BoNT/B and TeNT: residues adjacent to the site of scissile bond cleavage (cleavage region) and residues located within N-terminal and C-terminal regions relative to the cleavage region. Analysis of residues within the cleavage region showed that mutations at the P7, P4, P2, and P1' residues of VAMP-2 had the greatest inhibition of LC/B cleavage (> or =32-fold), whereas mutations at P7, P4, P1', and P2' residues of VAMP-2 had the greatest inhibition of LC/TeNT cleavage (> or =64-fold). Residues within the cleavage region influenced catalysis, whereas residues N-terminal and C-terminal to the cleavage region influenced binding affinity. Thus, BoNT/B and TeNT possess similar organization but have unique residues to recognize and cleave VAMP-2. These studies provide new insights into how the clostridial neurotoxins recognize their substrates.

  15. Botulinum Neurotoxin for Pain Management: Insights from Animal Models

    Directory of Open Access Journals (Sweden)

    Siro Luvisetto

    2010-12-01

    Full Text Available The action of botulinum neurotoxins (BoNTs at the neuromuscular junction has been extensively investigated and knowledge gained in this field laid the foundation for the use of BoNTs in human pathologies characterized by excessive muscle contractions. Although much more is known about the action of BoNTs on the peripheral system, growing evidence has demonstrated several effects also at the central level. Pain conditions, with special regard to neuropathic and intractable pain, are some of the pathological states that have been recently treated with BoNTs with beneficial effects. The knowledge of the action and potentiality of BoNTs utilization against pain, with emphasis for its possible use in modulation and alleviation of chronic pain, still represents an outstanding challenge for experimental research. This review highlights recent findings on the effects of BoNTs in animal pain models.

  16. Botulinum neurotoxin for pain management: insights from animal models.

    Science.gov (United States)

    Pavone, Flaminia; Luvisetto, Siro

    2010-12-01

    The action of botulinum neurotoxins (BoNTs) at the neuromuscular junction has been extensively investigated and knowledge gained in this field laid the foundation for the use of BoNTs in human pathologies characterized by excessive muscle contractions. Although much more is known about the action of BoNTs on the peripheral system, growing evidence has demonstrated several effects also at the central level. Pain conditions, with special regard to neuropathic and intractable pain, are some of the pathological states that have been recently treated with BoNTs with beneficial effects. The knowledge of the action and potentiality of BoNTs utilization against pain, with emphasis for its possible use in modulation and alleviation of chronic pain, still represents an outstanding challenge for experimental research. This review highlights recent findings on the effects of BoNTs in animal pain models.

  17. A monoclonal antibody based capture ELISA for botulinum neurotoxin serotype B: toxin detection in food

    Science.gov (United States)

    Botulism is a serious foodborne neuroparalyic disease caused by botulinum neurotoxin (BoNT) produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A-H) have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We repo...

  18. High-Throughput Screening Uncovers Novel Botulinum Neurotoxin Inhibitor Chemotypes.

    Science.gov (United States)

    Bompiani, Kristin M; Caglič, Dejan; Krutein, Michelle C; Benoni, Galit; Hrones, Morgan; Lairson, Luke L; Bian, Haiyan; Smith, Garry R; Dickerson, Tobin J

    2016-08-01

    Botulism is caused by potent and specific bacterial neurotoxins that infect host neurons and block neurotransmitter release. Treatment for botulism is limited to administration of an antitoxin within a short time window, before the toxin enters neurons. Alternatively, current botulism drug development targets the toxin light chain, which is a zinc-dependent metalloprotease that is delivered into neurons and mediates long-term pathology. Several groups have identified inhibitory small molecules, peptides, or aptamers, although no molecule has advanced to the clinic due to a lack of efficacy in advanced models. Here we used a homogeneous high-throughput enzyme assay to screen three libraries of drug-like small molecules for new chemotypes that modulate recombinant botulinum neurotoxin light chain activity. High-throughput screening of 97088 compounds identified numerous small molecules that activate or inhibit metalloprotease activity. We describe four major classes of inhibitory compounds identified, detail their structure-activity relationships, and assess their relative inhibitory potency. A previously unreported chemotype in any context of enzyme inhibition is described with potent submicromolar inhibition (Ki = 200-300 nM). Additional detailed kinetic analyses and cellular cytotoxicity assays indicate the best compound from this series is a competitive inhibitor with cytotoxicity values around 4-5 μM. Given the potency and drug-like character of these lead compounds, further studies, including cellular activity assays and DMPK analysis, are justified. PMID:27314875

  19. Evaluation of Lateral-Flow Clostridium botulinum Neurotoxin Detection Kits for Food Analysis

    OpenAIRE

    Sharma, Shashi K.; Eblen, Brian S.; Bull, Robert L.; Donald H. Burr; Whiting, Richard C.

    2005-01-01

    The suitability and sensitivity of two in vitro lateral-flow assays for detecting Clostridium botulinum neurotoxins (BoNTs) in an assortment of foods were evaluated. Toxin extraction and preparation methods for various liquid, solid, and high-fat-content foods were developed. The lateral-flow assays, one developed by the Naval Medical Research Center (Silver Spring, MD) and the other by Alexeter Technologies (Gaithersburg, MD), are based on the immunodetection of BoNT types A, B, and E. The a...

  20. Evaluation of adamantane hydroxamates as botulinum neurotoxin inhibitors: synthesis, crystallography, modeling, kinetic and cellular based studies

    OpenAIRE

    Šilhár, Peter; Silvaggi, Nicholas R; Pellett, Sabine; Čapková, Kateřina; Johnson, Eric A.; Allen, Karen N.; Janda, Kim D.

    2012-01-01

    Botulinum neurotoxins (BoNTs) are the most lethal biotoxins known to mankind and are responsible for the neuroparalytic disease botulism. Current treatments for botulinum poisoning are all protein based and thus have a limited window of treatment opportunity. Inhibition of the BoNT light chain protease (LC) has emerged as a therapeutic strategy for the treatment of botulism as it may provide an effective post exposure remedy. Using a combination of crystallographic and modeling studies a seri...

  1. Benzoquinones as inhibitors of botulinum neurotoxin serotype A.

    Science.gov (United States)

    Bremer, Paul T; Hixon, Mark S; Janda, Kim D

    2014-08-01

    Although botulinum neurotoxin serotype A (BoNT/A) is known for its use in cosmetics, it causes a potentially fatal illness, botulism, and can be used as a bioterror weapon. Many compounds have been developed that inhibit the BoNTA zinc-metalloprotease light chain (LC), however, none of these inhibitors have advanced to clinical trials. In this study, a fragment-based approach was implemented to develop novel covalent inhibitors of BoNT/A LC. First, electrophilic fragments were screened against BoNT/A LC, and benzoquinone (BQ) derivatives were found to be active. In kinetic studies, BQ compounds acted as irreversible inhibitors that presumably covalently modify cysteine 165 of BoNT/A LC. Although most BQ derivatives were highly reactive toward glutathione in vitro, a few compounds such as natural product naphthazarin displayed low thiol reactivity and good BoNT/A inhibition. In order to increase the potency of the BQ fragment, computational docking studies were employed to elucidate a scaffold that could bind to sites adjacent to Cys165 while positioning a BQ fragment at Cys165 for covalent modification; 2-amino-N-arylacetamides met these criteria and when linked to BQ displayed at least a 20-fold increase in activity to low μM IC₅₀ values. Unlike BQ alone, the linked-BQ compounds demonstrated only weak irreversible inhibition and therefore acted mainly as non-covalent inhibitors. Further kinetic studies revealed a mutual exclusivity of BQ covalent inactivation and competitive inhibitor binding to sites adjacent to Cys165, refuting the viability of the current strategy for developing more potent irreversible BoNT/A inhibitors. The highlights of this study include the discovery of BQ compounds as irreversible BoNT/A inhibitors and the rational design of low μM IC50 competitive inhibitors that depend on the BQ moiety for activity. PMID:24984937

  2. Antibody protection against botulinum neurotoxin intoxication in mice.

    Science.gov (United States)

    Cheng, Luisa W; Stanker, Larry H; Henderson, Thomas D; Lou, Jianlong; Marks, James D

    2009-10-01

    Adulteration of food or feed with any of the seven serotypes of botulinum neurotoxin (BoNT) is a potential bioterrorism concern. Currently, there is strong interest in the development of detection reagents, vaccines, therapeutics, and other countermeasures. A sensitive immunoassay for detecting BoNT serotype A (BoNT/A), based on monoclonal antibodies (MAbs) F1-2 and F1-40, has been developed and used in complex matrices. The epitope for F1-2 has been mapped to the heavy chain of BoNT/A, and the epitope of F1-40 has been mapped to the light chain. The ability of these MAbs to provide therapeutic protection against BoNT/A intoxication in mouse intravenous and oral intoxication models was tested. High dosages of individual MAbs protected mice well both pre- and postexposure to BoNT/A holotoxin. A combination therapy consisting of antibodies against both the light and heavy chains of the toxin, however, significantly increased protection, even at a lower MAb dosage. An in vitro peptide assay for measuring toxin activity showed that pretreatment of toxin with these MAbs did not block catalytic activity but instead blocked toxin entry into primary and cultured neuronal cells. The timing of antibody rescue in the mouse intoxication models revealed windows of opportunity for antibody therapeutic treatment that correlated well with the biologic half-life of the toxin in the serum. Knowledge of BoNT intoxication and antibody clearance in these mouse models and understanding of the pharmacokinetics of BoNT are invaluable for future development of antibodies and therapeutics against intoxication by BoNT.

  3. Botulinum Neurotoxins and Botulism: A Novel Therapeutic Approach

    Directory of Open Access Journals (Sweden)

    Wanpen Chaicumpa

    2011-05-01

    Full Text Available Specific treatment is not available for human botulism. Current remedial mainstay is the passive administration of polyclonal antibody to botulinum neurotoxin (BoNT derived from heterologous species (immunized animal or mouse hybridoma together with supportive and symptomatic management. The antibody works extracellularly, probably by blocking the binding of receptor binding (R domain to the neuronal receptors; thus inhibiting cellular entry of the holo-BoNT. The antibody cannot neutralize the intracellular toxin. Moreover, a conventional antibody with relatively large molecular size (150 kDa is not accessible to the enzymatic groove and, thus, cannot directly inhibit the BoNT zinc metalloprotease activity. Recently, a 15–20 kDa single domain antibody (VHH that binds specifically to light chain of BoNT serotype A was produced from a humanized-camel VH/VHH phage display library. The VHH has high sequence homology (>80% to the human VH and could block the enzymatic activity of the BoNT. Molecular docking revealed not only the interface binding between the VHH and the toxin but also an insertion of the VHH CDR3 into the toxin enzymatic pocket. It is envisaged that, by molecular linking the VHH to a cell penetrating peptide (CPP, the CPP-VHH fusion protein would be able to traverse the hydrophobic cell membrane into the cytoplasm and inhibit the intracellular BoNT. This presents a novel and safe immunotherapeutic strategy for botulism by using a cell penetrating, humanized-single domain antibody that inhibits the BoNT by means of a direct blockade of the groove of the menace enzyme.

  4. Proconvulsant actions of intrahippocampal botulinum neurotoxin B in the rat.

    Science.gov (United States)

    Bröer, S; Zolkowska, D; Gernert, M; Rogawski, M A

    2013-11-12

    Botulinum neurotoxins (BoNTs) may affect the excitability of brain circuits by inhibiting neurotransmitter release at central synapses. There is evidence that local delivery of BoNT serotypes A and E, which target SNAP-25, a component of the release machinery specific to excitatory synapses, can inhibit seizure generation. BoNT serotype B (BoNT/B) targets VAMP2, which is expressed in both excitatory and inhibitory terminals. Here we assessed the effects of unilateral intrahippocampal infusion of BoNT/B in the rat on intravenous pentylenetetrazol (PTZ) seizure thresholds, and on the expression of spontaneous behavioral and electrographic seizures. Infusion of BoNT/B (500 and 1,000 unit) by convection-enhanced delivery caused a reduction in myoclonic twitch and clonic seizure thresholds in response to intravenous PTZ beginning about 6 days after the infusion. Handling-evoked and spontaneous convulsive seizures were observed in many BoNT/B-treated animals but not in vehicle-treated controls. Spontaneous electrographic seizure discharges were recorded in the dentate gyrus of animals that received local BoNT/B infusion. In addition, there was an increased frequency of interictal epileptiform spikes and sharp waves at the same recording site. BoNT/B-treated animals also exhibited tactile hyperresponsivity in comparison with vehicle-treated controls. This is the first demonstration that BoNT/B causes a delayed proconvulsant action when infused into the hippocampus. Local infusion of BoNT/B could be useful as a focal epilepsy model.

  5. An Invisible Lethal Killer: Clostridium Botulinum and Botulinum Neurotoxins%一种致命的隐形杀手:肉毒杆菌与肉毒毒素

    Institute of Scientific and Technical Information of China (English)

    王景林

    2013-01-01

    2013年8月,因肉毒杆菌污染,问题奶粉再次进入公众视野.肉毒杆菌(Clostridium botulinum)是一种革兰阳性粗短杆菌,严格厌氧,有A~G7个亚型,每个亚型都可产生一种剧毒的大分子外毒素,即肉毒毒素(botulinum toxin).这种毒素可引起人和动物发生以松弛性麻痹为主症的肉毒中毒(botulism),虽然并不常见,但却是一种致命的中毒性疾病.本文综述了肉毒杆菌生物学特征、肉毒毒素结构与毒性、肉毒中毒临床表现以及检验鉴定等,并介绍了食品安全生产中肉毒杆菌与肉毒毒素的安全风险与控制措施.%The milkpowder contaminated by Clostridium botulinum once again go into the public view in August 2013.Clostridium botulinum,which is a strictly anaerobic gram-positive bacillus,is classified into seven main types from A through G according to exotoxins and they can produce antigenically distinct botulinum neurotoxins.It is a potent toxin that causes the most severe form of food poisoning with pronounced flaccid paralysis,called botulism.The review covers biological features of C.botulinum,structure and toxicity of toxin,clinical symptoms and identification of botulism as well as the risk and control of C.botulinum and toxins during food processing and preservation.

  6. Neuronal targeting, internalization, and biological activity of a recombinant atoxic derivative of botulinum neurotoxin A

    Science.gov (United States)

    Botulinum neurotoxins (BoNT) have the unique capacity to cross epithelial barriers, target neuromuscular junctions, and translocate active metalloprotease component to the cytosol of motor neurons. We have taken advantage of the molecular carriers responsible for this trafficking to create a family ...

  7. Zebrafish (Danio rerio) bioassay for visceral toxicosis of catfish and botulinum neurotoxin serotype E

    Science.gov (United States)

    Visceral toxicosis of catfish (VTC), a sporadic disease of cultured channel catfish (Ictalurus punctatus) often with high mortality, is caused by botulinum neurotoxin serotype E (BoNT/E). Presumptive diagnosis of VTC is based on characteristic clinical signs and lesions, and the production of these ...

  8. Botulinum Neurotoxin Serotype A Recognizes Its Protein Receptor SV2 by a Different Mechanism than Botulinum Neurotoxin B Synaptotagmin

    Science.gov (United States)

    Weisemann, Jasmin; Stern, Daniel; Mahrhold, Stefan; Dorner, Brigitte G.; Rummel, Andreas

    2016-01-01

    Botulinum neurotoxins (BoNTs) exhibit extraordinary potency due to their exquisite neurospecificity, which is achieved by dual binding to complex polysialo-gangliosides and synaptic vesicle proteins. The luminal domain 4 (LD4) of the three synaptic vesicle glycoprotein 2 isoforms, SV2A‐C, identified as protein receptors for the most relevant serotype BoNT/A, binds within the 50 kDa cell binding domain HC of BoNT/A. Here, we deciphered the BoNT/A‐SV2 interactions in more detail. In pull down assays, the binding of HCA to SV2-LD4 isoforms decreases from SV2C >> SV2A > SV2B. A binding constant of 200 nM was determined for BoNT/A to rat SV2C-LD4 in GST pull down assay. A similar binding constant was determined by surface plasmon resonance for HCA to rat SV2C and to human SV2C, the latter being slightly lower due to the substitution L563F in LD4. At pH 5, as measured in acidic synaptic vesicles, the binding constant of HCA to hSV2C is increased more than 10-fold. Circular dichroism spectroscopy reveals that the quadrilateral helix of SV2C-LD4 already exists in solution prior to BoNT/A binding. Hence, the BoNT/A‐SV2C interaction is of different nature compared to BoNT/B‐Syt-II. In particular, the preexistence of the quadrilateral β-sheet helix of SV2 and its pH-dependent binding to BoNT/A via backbone–backbone interactions constitute major differences. Knowledge of the molecular details of BoNT/A‐SV2 interactions drives the development of high affinity peptides to counteract BoNT/A intoxications or to capture functional BoNT/A variants in innovative detection systems for botulism diagnostic. PMID:27196927

  9. Botulinum Neurotoxin Serotype A Recognizes Its Protein Receptor SV2 by a Different Mechanism than Botulinum Neurotoxin B Synaptotagmin

    Directory of Open Access Journals (Sweden)

    Jasmin Weisemann

    2016-05-01

    Full Text Available Botulinum neurotoxins (BoNTs exhibit extraordinary potency due to their exquisite neurospecificity, which is achieved by dual binding to complex polysialo-gangliosides and synaptic vesicle proteins. The luminal domain 4 (LD4 of the three synaptic vesicle glycoprotein 2 isoforms, SV2A‐C, identified as protein receptors for the most relevant serotype BoNT/A, binds within the 50 kDa cell binding domain HC of BoNT/A. Here, we deciphered the BoNT/A‐SV2 interactions in more detail. In pull down assays, the binding of HCA to SV2-LD4 isoforms decreases from SV2C >> SV2A > SV2B. A binding constant of 200 nM was determined for BoNT/A to rat SV2C-LD4 in GST pull down assay. A similar binding constant was determined by surface plasmon resonance for HCA to rat SV2C and to human SV2C, the latter being slightly lower due to the substitution L563F in LD4. At pH 5, as measured in acidic synaptic vesicles, the binding constant of HCA to hSV2C is increased more than 10-fold. Circular dichroism spectroscopy reveals that the quadrilateral helix of SV2C-LD4 already exists in solution prior to BoNT/A binding. Hence, the BoNT/A‐SV2C interaction is of different nature compared to BoNT/B‐Syt-II. In particular, the preexistence of the quadrilateral β-sheet helix of SV2 and its pH-dependent binding to BoNT/A via backbone–backbone interactions constitute major differences. Knowledge of the molecular details of BoNT/A‐SV2 interactions drives the development of high affinity peptides to counteract BoNT/A intoxications or to capture functional BoNT/A variants in innovative detection systems for botulism diagnostic.

  10. Characterization and immunological activity of different forms of recombinant secreted Hc of botulinum neurotoxin serotype B products expressed in yeast

    OpenAIRE

    Liu, Bo; Shi, DanYang; Chang, Shaohong; Gong, Xin; Yu, YunZhou; Sun, Zhiwei; Wu, Jun

    2015-01-01

    The recombinant Hc proteins of botulinum neurotoxins and tetanus toxin are exclusively produced by intracellular heterologous expression in Pichia pastoris for use in subunit vaccines; the same Hc proteins produced by secreted heterologous expression are hyper-glycosylated and immunologically inert. Here, several different recombinant secreted Hc proteins of botulinum neurotoxin serotype B (BHc) were expressed in yeast and we characterized and assessed their immunological activity in detail. ...

  11. MOLECULAR-BIOLOGY OF CLOSTRIDIAL TOXINS - EXPRESSION OF MESSENGER-RNAS ENCODING TETANUS AND BOTULINUM NEUROTOXINS IN APLYSIA NEURONS

    NARCIS (Netherlands)

    MOCHIDA, S; POULAIN, B; EISEL, U; BINZ, T; KURAZONO, H; NIEMANN, H; TAUC, L

    1990-01-01

    mRNAs encoding the light chain of tetanus and botulinum neurotoxins were transcribed, in vitro, from the cloned and specifically truncated genes of Clostridium tetani and Clostridium botulinum, respectively, and injected into presynaptic identified cholinergic neurons of the buccal ganglia of Aplysi

  12. Application of high-density DNA resequencing microarray for detection and characterization of botulinum neurotoxin-producing clostridia.

    Directory of Open Access Journals (Sweden)

    Jessica Vanhomwegen

    Full Text Available BACKGROUND: Clostridium botulinum and related clostridia express extremely potent toxins known as botulinum neurotoxins (BoNTs that cause severe, potentially lethal intoxications in humans. These BoNT-producing bacteria are categorized in seven major toxinotypes (A through G and several subtypes. The high diversity in nucleotide sequence and genetic organization of the gene cluster encoding the BoNT components poses a great challenge for the screening and characterization of BoNT-producing strains. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we designed and evaluated the performances of a resequencing microarray (RMA, the PathogenId v2.0, combined with an automated data approach for the simultaneous detection and characterization of BoNT-producing clostridia. The unique design of the PathogenID v2.0 array allows the simultaneous detection and characterization of 48 sequences targeting the BoNT gene cluster components. This approach allowed successful identification and typing of representative strains of the different toxinotypes and subtypes, as well as the neurotoxin-producing C. botulinum strain in a naturally contaminated food sample. Moreover, the method allowed fine characterization of the different neurotoxin gene cluster components of all studied strains, including genomic regions exhibiting up to 24.65% divergence with the sequences tiled on the arrays. CONCLUSIONS/SIGNIFICANCE: The severity of the disease demands rapid and accurate means for performing risk assessments of BoNT-producing clostridia and for tracing potentials sources of contamination in outbreak situations. The RMA approach constitutes an essential higher echelon component in a diagnostics and surveillance pipeline. In addition, it is an important asset to characterise potential outbreak related strains, but also environment isolates, in order to obtain a better picture of the molecular epidemiology of BoNT-producing clostridia.

  13. Two-component signal transduction system CBO0787/CBO0786 represses transcription from botulinum neurotoxin promoters in Clostridium botulinum ATCC 3502.

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    Zhen Zhang

    2013-03-01

    Full Text Available Blocking neurotransmission, botulinum neurotoxin is the most poisonous biological substance known to mankind. Despite its infamy as the scourge of the food industry, the neurotoxin is increasingly used as a pharmaceutical to treat an expanding range of muscle disorders. Whilst neurotoxin expression by the spore-forming bacterium Clostridium botulinum appears tightly regulated, to date only positive regulatory elements, such as the alternative sigma factor BotR, have been implicated in this control. The identification of negative regulators has proven to be elusive. Here, we show that the two-component signal transduction system CBO0787/CBO0786 negatively regulates botulinum neurotoxin expression. Single insertional inactivation of cbo0787 encoding a sensor histidine kinase, or of cbo0786 encoding a response regulator, resulted in significantly elevated neurotoxin gene expression levels and increased neurotoxin production. Recombinant CBO0786 regulator was shown to bind to the conserved -10 site of the core promoters of the ha and ntnh-botA operons, which encode the toxin structural and accessory proteins. Increasing concentration of CBO0786 inhibited BotR-directed transcription from the ha and ntnh-botA promoters, demonstrating direct transcriptional repression of the ha and ntnh-botA operons by CBO0786. Thus, we propose that CBO0786 represses neurotoxin gene expression by blocking BotR-directed transcription from the neurotoxin promoters. This is the first evidence of a negative regulator controlling botulinum neurotoxin production. Understanding the neurotoxin regulatory mechanisms is a major target of the food and pharmaceutical industries alike.

  14. Isolation and Quantification of Botulinum Neurotoxin From Complex Matrices Using the BoTest Matrix Assays

    OpenAIRE

    Dunning, F. Mark; Piazza, Timothy M.; Zeytin, Füsûn N.; Tucker, Ward C.

    2014-01-01

    Accurate detection and quantification of botulinum neurotoxin (BoNT) in complex matrices is required for pharmaceutical, environmental, and food sample testing. Rapid BoNT testing of foodstuffs is needed during outbreak forensics, patient diagnosis, and food safety testing while accurate potency testing is required for BoNT-based drug product manufacturing and patient safety. The widely used mouse bioassay for BoNT testing is highly sensitive but lacks the precision and throughput needed for ...

  15. Algal chloroplast produced camelid VHH antitoxins are capable of neutralizing botulinum neurotoxin

    OpenAIRE

    Daniel J Barrera; Rosenberg, Julian N.; Chiu, Joanna G.; Chang, Yung-Nien; Debatis, Michelle; Ngoi, Soo-Mun; Chang, John T.; Shoemaker, Charles B.; George A Oyler; Mayfield, Stephen P

    2014-01-01

    We have produced three antitoxins consisting of the variable domains of camelid heavy chain-only antibodies (VHH) by expressing the genes in the chloroplast of green algae. These antitoxins accumulate as soluble proteins capable of binding and neutralizing botulinum neurotoxin. Furthermore, they accumulate at up to 5% total soluble protein, sufficient expression to easily produce these antitoxins at scale from algae. The genes for the three different antitoxins were transformed into Chlamydom...

  16. Treatment of Chronic Migraine with Focus on Botulinum Neurotoxins

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    Sara M. Schaefer

    2015-07-01

    Full Text Available Migraine is the most common neurological disorder, and contributes to disability and large healthcare costs in the United States and the world. The treatment of migraine until recently has focused on medications, both abortive and prophylactic, but treatment of chronic migraine has been revolutionized with the introduction of botulinum toxin injection therapy. In this review, we explore the current understanding of migraine pathophysiology, and the evolution of the use of botulinum toxin therapy including proposed pathophysiological mechanisms through animal data. We also discuss the similarities and differences between three injection techniques.

  17. Re-Assembled Botulinum Neurotoxin Inhibits CNS Functions without Systemic Toxicity

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    Bazbek Davletov

    2011-03-01

    Full Text Available The therapeutic potential of botulinum neurotoxin type A (BoNT/A has recently been widely recognized. BoNT/A acts to silence synaptic transmission via specific proteolytic cleavage of an essential neuronal protein, SNAP25. The advantages of BoNT/A-mediated synaptic silencing include very long duration, high potency and localized action. However, there is a fear of possible side-effects of BoNT/A due to its diffusible nature which may lead to neuromuscular blockade away from the injection site. We recently developed a “protein-stapling” technology which allows re-assembly of BoNT/A from two separate fragments. This technology allowed, for the first time, safe production of this popular neuronal silencing agent. Here we evaluated the re-assembled toxin in several CNS assays and assessed its systemic effects in an animal model. Our results show that the re-assembled toxin is potent in inhibiting CNS function at 1 nM concentration but surprisingly does not exhibit systemic toxicity after intraperitoneal injection even at 200 ng/kg dose. This shows that the re-assembled toxin represents a uniquely safe tool for neuroscience research and future medical applications.

  18. The destructive effect of botulinum neurotoxins on the SNARE protein: SNAP-25 and synaptic membrane fusion

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    Bin Lu

    2015-06-01

    Full Text Available Synaptic exocytosis requires the assembly of syntaxin 1A and SNAP-25 on the plasma membrane and synaptobrevin 2 (VAMP2 on the vesicular membrane to bridge the two opposite membranes. It is believed that the three SNARE proteins assemble in steps along the dynamic assembly pathway. The C-terminus of SNAP-25 is known to be the target of botulinum neurotoxins (BoNT/A and BoNT/E that block neurotransmitters release in vivo. In this study, we employed electron paramagnetic resonance (EPR spectroscopy to investigate the conformation of the SNAP-25 C-terminus in binary and ternary SNARE complexes. The fluorescence lipid mixing assay shows that the C-terminal of SNAP-25 is essential for membrane fusion, and that the truncated SNAP-25 mutants cleaved by BoNT/A and BoNT/E display different inhibition effects on membrane fusion: SNAP-25E (Δ26 abolishes the fusion activity of the SNARE complex, while SNAP-25A (Δ9 loses most of its function, although it can still form a SDS-resistant SNARE complex as the wild-type SNAP-25. CW-EPR spectra validate the unstable structures of the SNARE complex formed by SNAP-25 mutants. We propose that the truncated SNAP-25 mutants will disrupt the assembly of the SNARE core complex, and then inhibit the synaptic membrane fusion accordingly.

  19. VAMP/synaptobrevin cleavage by tetanus and botulinum neurotoxins is strongly enhanced by acidic liposomes.

    Science.gov (United States)

    Caccin, Paola; Rossetto, Ornella; Rigoni, Michela; Johnson, Eric; Schiavo, Giampietro; Montecucco, Cesare

    2003-05-01

    Tetanus and botulinum neurotoxins (TeNT and BoNTs) block neuroexocytosis via specific cleavage and inactivation of SNARE proteins. Such activity is exerted by the N-terminal 50 kDa light chain (L) domain, which is a zinc-dependent endopeptidase. TeNT, BoNT/B, /D, /F and /G cleave vesicle associated membrane protein (VAMP), a protein of the neurotransmitter-containing small synaptic vesicles, at different single peptide bonds. Since the proteolytic activity of these metalloproteases is higher on native VAMP inserted in synaptic vesicles than on recombinant VAMP, we have investigated the influence of liposomes of different lipid composition on this activity. We found that the rate of VAMP cleavage with all neurotoxins tested here is strongly enhanced by negatively charged lipid mixtures. This effect is at least partially due to the binding of the metalloprotease to the lipid membranes, with electrostatic interactions playing an important role.

  20. Botulinum Neurotoxins Can Enter Cultured Neurons Independent of Synaptic Vesicle Recycling

    OpenAIRE

    Pellett, Sabine; Tepp, William H.; Jacob M Scherf; Eric A Johnson

    2015-01-01

    Botulinum neurotoxins (BoNTs) are the causative agent of the severe and long-lasting disease botulism. At least seven different serotypes of BoNTs (denoted A-G) have been described. All BoNTs enter human or animal neuronal cells via receptor mediated endocytosis and cleave cytosolic SNARE proteins, resulting in a block of synaptic vesicle exocytosis, leading to the flaccid paralysis characteristic of botulism. Previous data have indicated that once a neuronal cell has been intoxicated by a Bo...

  1. Pre-Clinical Study of a Novel Recombinant Botulinum Neurotoxin Derivative Engineered for Improved Safety.

    Science.gov (United States)

    Vazquez-Cintron, Edwin; Tenezaca, Luis; Angeles, Christopher; Syngkon, Aurelia; Liublinska, Victoria; Ichtchenko, Konstantin; Band, Philip

    2016-01-01

    Cyto-012 is a recombinant derivative of Botulinum neurotoxin Type A (BoNT/A). It primarily differs from wild type (wt) BoNT/A1 in that it incorporates two amino acid substitutions in the catalytic domain of the light chain (LC) metalloprotease (E224 > A and Y366 > A), designed to provide a safer clinical profile. Cyto-012 is specifically internalized into rat cortical and hippocampal neurons, and cleaves Synaptosomal-Associated Protein 25 (SNAP-25), the substrate of wt BoNT/A, but exhibits slower cleavage kinetics and therefore requires a higher absolute dose to exhibit pharmacologic activity. The pharmacodynamics of Cyto-012 and wt BoNT/A have similar onset and duration of action using the Digital Abduction Assay (DAS). Intramuscular LD50 values for Cyto-012 and wt BoNT/A respectively, were 0.63 ug (95% CI = 0.61, 0.66) and 6.22 pg (95% CI = 5.42, 7.02). ED50 values for Cyto-012 and wt BoNT/A were respectively, 0.030 ug (95% CI = 0.026, 0.034) and 0.592 pg (95% CI = 0.488, 0.696). The safety margin (intramuscular LD50/ED50 ratio) for Cyto-012 was found to be improved 2-fold relative to wt BoNT/A (p < 0.001). The DAS response to Cyto-012 was diminished when a second injection was administered 32 days after the first. These data suggest that the safety margin of BoNT/A can be improved by modulating their activity towards SNAP-25.

  2. Algal chloroplast produced camelid VH H antitoxins are capable of neutralizing botulinum neurotoxin.

    Science.gov (United States)

    Barrera, Daniel J; Rosenberg, Julian N; Chiu, Joanna G; Chang, Yung-Nien; Debatis, Michelle; Ngoi, Soo-Mun; Chang, John T; Shoemaker, Charles B; Oyler, George A; Mayfield, Stephen P

    2015-01-01

    We have produced three antitoxins consisting of the variable domains of camelid heavy chain-only antibodies (VH H) by expressing the genes in the chloroplast of green algae. These antitoxins accumulate as soluble proteins capable of binding and neutralizing botulinum neurotoxin. Furthermore, they accumulate at up to 5% total soluble protein, sufficient expression to easily produce these antitoxins at scale from algae. The genes for the three different antitoxins were transformed into Chlamydomonas reinhardtii chloroplasts and their products purified from algae lysates and assayed for in vitro biological activity using toxin protection assays. The produced antibody domains bind to botulinum neurotoxin serotype A (BoNT/A) with similar affinities as camelid antibodies produced in Escherichia coli, and they are similarly able to protect primary rat neurons from intoxication by BoNT/A. Furthermore, the camelid antibodies were produced in algae without the use of solubilization tags commonly employed in E. coli. These camelid antibody domains are potent antigen-binding proteins and the heterodimer fusion protein containing two VH H domains was capable of neutralizing BoNT/A at near equimolar concentrations with the toxin. Intact antibody domains were detected in the gastrointestinal (GI) tract of mice treated orally with antitoxin-producing microalgae. These findings support the use of orally delivered antitoxins produced in green algae as a novel treatment for botulism.

  3. Mass Spectrometric Detection of Botulinum Neurotoxin by Measuring its Activity in Serum and Milk

    Science.gov (United States)

    Kalb, Suzanne R.; Pirkle, James L.; Barr, John R.

    Botulinum neurotoxins (BoNTs) are bacterial protein toxins which are considered likely agents for bioterrorism due to their extreme toxicity and high availability. A new mass spectrometry based assay called Endopep MS detects and defines the toxin serotype in clinical and food matrices via toxin activity upon a peptide substrate which mimics the toxin's natural target. Furthermore, the subtype of the toxin is differentiated by employing mass spectrometry based proteomic techniques on the same sample. The Endopep-MS assay selectively detects active BoNT and defines the serotype faster and with sensitivity greater than the mouse bioassay. One 96-well plate can be analyzed in under 7 h. On higher level or "hot" samples, the subtype can then be differentiated in less than 2 h with no need for DNA.

  4. A Monoclonal Antibody Based Capture ELISA for Botulinum Neurotoxin Serotype B: Toxin Detection in Food

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    Larry H. Stanker

    2013-11-01

    Full Text Available Botulism is a serious foodborne neuroparalytic disease, caused by botulinum neurotoxin (BoNT, produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A–H have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We report here a group of serotype B specific monoclonal antibodies (mAbs capable of binding toxin under physiological conditions. Thus, they serve as capture antibodies for a sandwich (capture ELISA. The antibodies were generated using recombinant peptide fragments corresponding to the receptor-binding domain of the toxin heavy chain as immunogen. Their binding properties suggest that they bind a complex epitope with dissociation constants (KD’s for individual antibodies ranging from 10 to 48 × 10−11 M. Assay performance for all possible combinations of capture-detector antibody pairs was evaluated and the antibody pair resulting in the lowest level of detection (L.O.D., ~20 pg/mL was determined. Toxin was detected in spiked dairy samples with good recoveries at concentrations as low as 0.5 pg/mL and in ground beef samples at levels as low as 2 ng/g. Thus, the sandwich ELISA described here uses mAb for both the capture and detector antibodies (binding different epitopes on the toxin molecule and readily detects toxin in those food samples tested.

  5. Identification of a Unique Ganglioside Binding Loop within Botulinum Neurotoxins C and D-SA

    Energy Technology Data Exchange (ETDEWEB)

    Karalewitz, Andrew P.-A.; Kroken, Abby R.; Fu, Zhuji; Baldwin, Michael R.; Kim, Jung-Ja P.; Barbieri, Joseph T. (MCW); (Missouri)

    2010-09-22

    The botulinum neurotoxins (BoNTs) are the most potent protein toxins for humans. There are seven serotypes of BoNTs (A-G) based on a lack of cross antiserum neutralization. BoNTs utilize gangliosides as components of the host receptors for binding and entry into neurons. Members of BoNT/C and BoNT/D serotypes include mosaic toxins that are organized in D/C and C/D toxins. One D/C mosaic toxin, BoNT/D-South Africa (BoNT/D-SA), was not fully neutralized by immunization with BoNT serotype C or D, which stimulated this study. Here the crystal structures of the receptor binding domains of BoNT/C, BoNT/D, and BoNT/D-SA are presented. Biochemical and cell binding studies show that BoNT/C and BoNT/D-SA possess unique mechanisms for ganglioside binding. These studies provide new information about how the BoNTs can enter host cells as well as a basis for understanding the immunological diversity of these neurotoxins.

  6. Accelerated neuronal cell recovery from Botulinum neurotoxin intoxication by targeted ubiquitination.

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    Chueh-Ling Kuo

    Full Text Available Botulinum neurotoxin (BoNT, a Category A biodefense agent, delivers a protease to motor neuron cytosol that cleaves one or more soluble NSF attachment protein receptors (SNARE proteins involved in neurotransmission to cause a flaccid paralysis. No antidotes exist to reverse symptoms of BoNT intoxication so severely affected patients require artificial respiration with prolonged intensive care. Time to recovery depends on toxin serotype because the intraneuronal persistence of the seven known BoNT serotypes varies widely from days to many months. Our therapeutic antidote strategy is to develop 'targeted F-box' (TFB agents that target the different intraneuronal BoNT proteases for accelerated degradation by the ubiquitin proteasome system (UPS, thus promoting rapid recovery from all serotypes. These agents consist of a camelid heavy chain-only V(H (VHH domain specific for a BoNT protease fused to an F-box domain recognized by an intraneuronal E3-ligase. A fusion protein containing the 14 kDa anti-BoNT/A protease VHH, ALcB8, joined to a 15 kDa F-box domain region of TrCP (D5 was sufficient to cause increased ubiquitination and accelerate turnover of the targeted BoNT/A protease within neurons. Neuronal cells expressing this TFB, called D5-B8, were also substantially resistant to BoNT/A intoxication and recovered from intoxication at least 2.5 fold quicker than control neurons. Fusion of D5 to a VHH specific for BoNT/B protease (BLcB10 led to accelerated turnover of the targeted protease within neurons, thus demonstrating the modular nature of these therapeutic agents and suggesting that development of similar therapeutic agents specific to all botulinum serotypes should be readily achievable.

  7. Accelerated neuronal cell recovery from Botulinum neurotoxin intoxication by targeted ubiquitination.

    Science.gov (United States)

    Kuo, Chueh-Ling; Oyler, George A; Shoemaker, Charles B

    2011-01-01

    Botulinum neurotoxin (BoNT), a Category A biodefense agent, delivers a protease to motor neuron cytosol that cleaves one or more soluble NSF attachment protein receptors (SNARE) proteins involved in neurotransmission to cause a flaccid paralysis. No antidotes exist to reverse symptoms of BoNT intoxication so severely affected patients require artificial respiration with prolonged intensive care. Time to recovery depends on toxin serotype because the intraneuronal persistence of the seven known BoNT serotypes varies widely from days to many months. Our therapeutic antidote strategy is to develop 'targeted F-box' (TFB) agents that target the different intraneuronal BoNT proteases for accelerated degradation by the ubiquitin proteasome system (UPS), thus promoting rapid recovery from all serotypes. These agents consist of a camelid heavy chain-only V(H) (VHH) domain specific for a BoNT protease fused to an F-box domain recognized by an intraneuronal E3-ligase. A fusion protein containing the 14 kDa anti-BoNT/A protease VHH, ALcB8, joined to a 15 kDa F-box domain region of TrCP (D5) was sufficient to cause increased ubiquitination and accelerate turnover of the targeted BoNT/A protease within neurons. Neuronal cells expressing this TFB, called D5-B8, were also substantially resistant to BoNT/A intoxication and recovered from intoxication at least 2.5 fold quicker than control neurons. Fusion of D5 to a VHH specific for BoNT/B protease (BLcB10) led to accelerated turnover of the targeted protease within neurons, thus demonstrating the modular nature of these therapeutic agents and suggesting that development of similar therapeutic agents specific to all botulinum serotypes should be readily achievable. PMID:21629663

  8. Botulinum neurotoxins for the treatment of focal dystonias: Review of rating tools used in clinical trials.

    Science.gov (United States)

    Del Sorbo, Francesca; Albanese, Alberto

    2015-12-01

    Botulinum neurotoxins (BoNTs) are used to achieve therapeutic benefit in focal dystonia. An expert panel recently reviewed published evidence on the efficacy of BoNTs for the treatment of focal dystonias and produced recommendations for clinical practice. Another panel reviewed the clinimetric properties of rating scales for dystonia and produced recommendations for current usage and future directions. Considering that the strength of evidence derives not only from the quality of the study design, but also from usage of validated outcome measures, we combined the information provided by these two recent reviews and assessed the appropriateness of the rating instruments used in clinical trials on BoNT treatment in focal dystonia. Data sources included all the publications on BoNT treatment for focal dystonias reviewed by the recent evidence-based analysis. We reviewed all rating instruments used to assess primary and secondary outcome following BoNT treatment. The publications were allocated into five topics according to the focal dystonia type reviewed in the meta-analysis: blepharospasm, oromandibular dystonia, cervical dystonia, upper limb dystonia, and laryngeal dystonia. For each topic, papers were divided, according to the terminology used in the meta-analysis, into placebo-controlled, active comparator and methodological or uncontrolled. For each topic we identified the rating tools used in each study class and annotated which were the mostly used in each focal dystonia type. Outcome measures included tools related to motor and non-motor features, such as pain and depression, and functional as well as health-related quality of life features. Patient- and investigator-reported outcomes were also included. Rating instruments were classified as recommended, suggested, listed or not included, based on recommendations produced by the rating scale task force. Both primary and secondary outcome measures were assessed. As a final step we compared current practice, as

  9. Plastic enzyme-linked immunosorbent assays (ELISA)-on-a-chip biosensor for botulinum neurotoxin A.

    Science.gov (United States)

    Han, Seung-Mok; Cho, Joung-Hwan; Cho, Il-Hoon; Paek, Eui-Hwan; Oh, Hee-Bok; Kim, Bong-Su; Ryu, Chunsun; Lee, Kyunghee; Kim, Young-Kee; Paek, Se-Hwan

    2007-03-21

    A plastic ELISA-on-a-chip (EOC) employing the concept of cross-flow immuno-chromatographic analysis was applied to the measurement of botulinum neurotoxin A (BoNT/A) as agent for bio-terrorism. Two monoclonal antibodies specific to the heavy chain of the toxin were raised and identified to form sandwich binding complexes as the pair with the analyte. For the construction of an immuno-strip, one was utilized as the capture antibody immobilized onto nitrocellulose membrane and the other as the detection coupled to an enzyme, horseradish peroxidase. The two plates of EOC used in this study were fabricated by injection molding of polycarbonate to improve the reproducibility of manufacture and, after inclusion of the immuno-strip, bonded using a UV-sensitive adhesive. Under optimal conditions of analysis, the chip produced a color signal in proportion to the analyte dose and the signal was quantified using a detector equipped with a digital camera. From the dose-response curve, the detection limit of BoNT/A was 2.0 ng mL(-1), approximately five times more sensitive than a commercial-version detection kit employing colloidal gold tracer.

  10. Inhibiting oral intoxication of botulinum neurotoxin A complex by carbohydrate receptor mimics.

    Science.gov (United States)

    Lee, Kwangkook; Lam, Kwok-Ho; Kruel, Anna-Magdalena; Mahrhold, Stefan; Perry, Kay; Cheng, Luisa W; Rummel, Andreas; Jin, Rongsheng

    2015-12-01

    Botulinum neurotoxins (BoNTs) cause the disease botulism manifested by flaccid paralysis that could be fatal to humans and animals. Oral ingestion of the toxin with contaminated food is one of the most common routes for botulism. BoNT assembles with several auxiliary proteins to survive in the gastrointestinal tract and is subsequently transported through the intestinal epithelium into the general circulation. Several hemagglutinin proteins form a multi-protein complex (HA complex) that recognizes host glycans on the intestinal epithelial cell surface to facilitate BoNT absorption. Blocking carbohydrate binding to the HA complex could significantly inhibit the oral toxicity of BoNT. Here, we identify lactulose, a galactose-containing non-digestible sugar commonly used to treat constipation, as a prototype inhibitor against oral BoNT/A intoxication. As revealed by a crystal structure, lactulose binds to the HA complex at the same site where the host galactose-containing carbohydrate receptors bind. In vitro assays using intestinal Caco-2 cells demonstrated that lactulose inhibits HA from compromising the integrity of the epithelial cell monolayers and blocks the internalization of HA. Furthermore, co-administration of lactulose significantly protected mice against BoNT/A oral intoxication in vivo. Taken together, these data encourage the development of carbohydrate receptor mimics as a therapeutic intervention to prevent BoNT oral intoxication.

  11. The Receptor Binding Domain of Botulinum Neurotoxin Stereotype C Binds Phosphoinositides

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yanfeng; Varnum, Susan M.

    2012-03-01

    Botulinum neurotoxins (BoNTs) are the most toxic proteins known for humans and animals with an extremely low LD50 of {approx} 1 ng/kg. BoNTs generally require a protein and a ganglioside on the cell membrane surface for binding, which is known as a 'dual receptor' mechanism for host intoxication. Recent studies have suggested that in addition to gangliosides, other membrane lipids such as phosphoinositides may be involved in the interactions with the receptor binding domain (HCR) of BoNTs for better membrane penetration. Here, using two independent lipid-binding assays, we tested the interactions of BoNT/C-HCR with lipids in vitro. BoNT/C-HCR was found to bind negatively charged phospholipids, preferentially phosphoinositides. Additional interactions to phosphoinositides may help BoNT/C bind membrane more tightly and transduct signals for subsequent steps of intoxication. Our results provide new insights into the mechanisms of host cell membrane recognition by BoNTs.

  12. Clinical Uses of Botulinum Neurotoxins: Current Indications, Limitations and Future Developments

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    Sheng Chen

    2012-10-01

    Full Text Available Botulinum neurotoxins (BoNTs cause flaccid paralysis by interfering with vesicle fusion and neurotransmitter release in the neuronal cells. BoNTs are the most widely used therapeutic proteins. BoNT/A was approved by the U.S. FDA to treat strabismus, blepharospam, and hemificial spasm as early as 1989 and then for treatment of cervical dystonia, glabellar facial lines, axillary hyperhidrosis, chronic migraine and for cosmetic use. Due to its high efficacy, longevity of action and satisfactory safety profile, it has been used empirically in a variety of ophthalmological, gastrointestinal, urological, orthopedic, dermatological, secretory, and painful disorders. Currently available BoNT therapies are limited to neuronal indications with the requirement of periodic injections resulting in immune-resistance for some indications. Recent understanding of the structure-function relationship of BoNTs prompted the engineering of novel BoNTs to extend therapeutic interventions in non-neuronal systems and to overcome the immune-resistance issue. Much research still needs to be done to improve and extend the medical uses of BoNTs.

  13. Iterative structure-based peptide-like inhibitor design against the botulinum neurotoxin serotype A.

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    Jorge E Zuniga

    Full Text Available The botulinum neurotoxin serotype A light chain (BoNT/A LC protease is the catalytic component responsible for the neuroparalysis that is characteristic of the disease state botulism. Three related peptide-like molecules (PLMs were designed using previous information from co-crystal structures, synthesized, and assayed for in vitro inhibition against BoNT/A LC. Our results indicate these PLMS are competitive inhibitors of the BoNT/A LC protease and their K(i values are in the nM-range. A co-crystal structure for one of these inhibitors was determined and reveals that the PLM, in accord with the goals of our design strategy, simultaneously involves both ionic interactions via its P1 residue and hydrophobic contacts by means of an aromatic group in the P2' position. The PLM adopts a helical conformation similar to previously determined co-crystal structures of PLMs, although there are also major differences to these other structures such as contacts with specific BoNT/A LC residues. Our structure further demonstrates the remarkable plasticity of the substrate binding cleft of the BoNT/A LC protease and provides a paradigm for iterative structure-based design and development of BoNT/A LC inhibitors.

  14. Recommended Immunological Strategies to Screen for Botulinum Neurotoxin-Containing Samples

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    Stéphanie Simon

    2015-11-01

    Full Text Available Botulinum neurotoxins (BoNTs cause the life-threatening neurological illness botulism in humans and animals and are divided into seven serotypes (BoNT/A–G, of which serotypes A, B, E, and F cause the disease in humans. BoNTs are classified as “category A” bioterrorism threat agents and are relevant in the context of the Biological Weapons Convention. An international proficiency test (PT was conducted to evaluate detection, quantification and discrimination capabilities of 23 expert laboratories from the health, food and security areas. Here we describe three immunological strategies that proved to be successful for the detection and quantification of BoNT/A, B, and E considering the restricted sample volume (1 mL distributed. To analyze the samples qualitatively and quantitatively, the first strategy was based on sensitive immunoenzymatic and immunochromatographic assays for fast qualitative and quantitative analyses. In the second approach, a bead-based suspension array was used for screening followed by conventional ELISA for quantification. In the third approach, an ELISA plate format assay was used for serotype specific immunodetection of BoNT-cleaved substrates, detecting the activity of the light chain, rather than the toxin protein. The results provide guidance for further steps in quality assurance and highlight problems to address in the future.

  15. Neutralization of botulinum neurotoxin by a human monoclonal antibody specific for the catalytic light chain.

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    Sharad P Adekar

    Full Text Available BACKGROUND: Botulinum neurotoxins (BoNT are a family of category A select bioterror agents and the most potent biological toxins known. Cloned antibody therapeutics hold considerable promise as BoNT therapeutics, but the therapeutic utility of antibodies that bind the BoNT light chain domain (LC, a metalloprotease that functions in the cytosol of cholinergic neurons, has not been thoroughly explored. METHODS AND FINDINGS: We used an optimized hybridoma method to clone a fully human antibody specific for the LC of serotype A BoNT (BoNT/A. The 4LCA antibody demonstrated potent in vivo neutralization when administered alone and collaborated with an antibody specific for the HC. In Neuro-2a neuroblastoma cells, the 4LCA antibody prevented the cleavage of the BoNT/A proteolytic target, SNAP-25. Unlike an antibody specific for the HC, the 4LCA antibody did not block entry of BoNT/A into cultured cells. Instead, it was taken up into synaptic vesicles along with BoNT/A. The 4LCA antibody also directly inhibited BoNT/A catalytic activity in vitro. CONCLUSIONS: An antibody specific for the BoNT/A LC can potently inhibit BoNT/A in vivo and in vitro, using mechanisms not previously associated with BoNT-neutralizing antibodies. Antibodies specific for BoNT LC may be valuable components of an antibody antidote for BoNT exposure.

  16. Camelid-derived heavy-chain nanobody against Clostridium botulinum neurotoxin E in Pichia pastoris.

    Science.gov (United States)

    Baghban, Roghayyeh; Gargari, Seyed Latif Mousavi; Rajabibazl, Masoumeh; Nazarian, Shahram; Bakherad, Hamid

    2016-01-01

    Botulinum neurotoxins (BoNTs) result in severe and often fatal disease, botulism. Common remedial measures such as equine antitoxin and human botulism immunoglobulin in turn are problematic and time-consuming. Therefore, diagnosis and therapy of BoNTs are vital. The variable domain of heavy-chain antibodies (VHH) has unique features, such as the ability to identify and bind specifically to target epitopes and ease of production in bacteria and yeast. The Pichia pastoris is suitable for expression of recombinant antibody fragments. Disulfide bond formation and correct folds of protein with a high yield are some of the advantages of this eukaryotic host. In this study, we have expressed and purified the camelid VHH against BoNT/E in P. pastoris. The final yield of P. pastoris-expressed antibody was estimated to be 16 mg/l, which is higher than that expressed by Escherichia coli. The nanobody expressed in P. pastoris neutralized 4LD50 of the BoNT/E upon i.p. injection in 25% of mice. The nanobody expressed in E. coli extended the mice's survival to 1.5-fold compared to the control. This experiment indicated that the quality of expressed protein in the yeast is superior to that of the bacterial expression. Favorable protein folding by P. pastoris seems to play a role in its better toxin-binding property. PMID:24673401

  17. Use of Monoclonal Antibodies in the Sensitive Detection and Neutralization of Botulinum Neurotoxin Serotype B

    Directory of Open Access Journals (Sweden)

    Luisa W. Cheng

    2015-11-01

    Full Text Available Botulinum neurotoxins (BoNT are some of nature’s most potent toxins. Due to potential food contamination, and bioterrorism concerns, the development of detection reagents, therapeutics and countermeasures are of urgent interest. Recently, we have developed a sensitive electrochemiluminescent (ECL immunoassay for BoNT/B, using monoclonal antibodies (mAbs MCS6-27 and anti-BoNT/B rabbit polyclonal antibodies as the capture and detector. The ECL assay detected as little as 1 pg/mL BoNT/B in the buffer matrix, surpassing the detection sensitivities of the gold standard mouse bioassays. The ECL assay also allowed detection of BoNT/B in sera matrices of up to 100% sera with negligible matrix effects. This highly-sensitive assay allowed the determination of the biological half-lives of BoNT/B holotoxin in vivo. We further tested the toxin neutralization potential of our monoclonal antibodies using the mouse systemic and oral intoxication models. A combination of mAbs protected mice in both pre- and post-exposure models to lethal doses of BoNT/B. MAbs were capable of increasing survival of animals when administered even 10 h post-intoxication in an oral model, suggesting a likely time for BoNT/B complexes to reach the blood stream. More sensitive detection assays and treatments against BoNT intoxication will greatly enhance efforts to combat botulism.

  18. Mode of VAMP substrate recognition and inhibition of Clostridium botulinum neurotoxin F.

    Science.gov (United States)

    Agarwal, Rakhi; Schmidt, James J; Stafford, Robert G; Swaminathan, Subramanyam

    2009-07-01

    Clostridium botulinum neurotoxins (BoNTs) cleave neuronal proteins responsible for neurotransmitter release, causing the neuroparalytic disease botulism. BoNT serotypes B, D, F and G cleave and inactivate vesicle-associated membrane protein (VAMP), each at a unique peptide bond. The specificity of BoNTs depends on the mode of substrate recognition. We have investigated the mechanism of substrate recognition of BoNT F by determining the crystal structures of its complex with two substrate-based inhibitors, VAMP 22-58/Gln58D-cysteine and 27-58/Gln58D-cysteine. The inhibitors bind to BoNT F in the canonical direction (as seen for BoNTs A and E substrates) but are positioned specifically via three major exosites away from the active site. The cysteine sulfur of the inhibitors interacts with the zinc and exists as sulfinic acid in the inhibitor VAMP 27-58/Gln58D-cysteine. Arg133 and Arg171, which form part of two separate exosites, are crucial for substrate binding and catalysis.

  19. Molecular components required for resting and stimulated endocytosis of botulinum neurotoxins by glutamatergic and peptidergic neurons.

    Science.gov (United States)

    Meng, Jianghui; Wang, Jiafu; Lawrence, Gary W; Dolly, J Oliver

    2013-08-01

    Proteins responsible for basal and stimulated endocytosis in nerves containing small clear synaptic vesicles (SCSVs) or large dense-core vesicles (LDCVs) are revealed herein, using probes that exploit surface-exposed vesicle proteins as acceptors for internalization. Basal uptake of botulinum neurotoxins (BoNTs) by both SCSV-releasing cerebellar granule neurons (CGNs) and LDCV-enriched trigeminal ganglionic neurons (TGNs) was found to require protein acceptors and acidic compartments. In addition, dynamin, clathrin, adaptor protein complex-2 (AP2), and amphiphysin contribute to the depolarization-evoked entry. For fast recycling of SCSVs, knockdown and knockout strategies demonstrated that CGNs use predominantly dynamin 1, whereas isoform 2 and, to a smaller extent, isoform 3 support a less rapid mode of stimulated endocytosis. Accordingly, proximity ligation assay confirmed that dynamin 1 and 2 colocalize with amphiphysin 1 in CGNs, and the latter copurified with both dynamins from cell extracts. In contrast, LDCV-releasing TGNs preferentially employ dynamins 2 and 3 and amphiphysin 1 for evoked endocytosis and lack the fast phase. Hence, stimulation recruits dynamin, clathrin, AP2, and amphiphysin to augment BoNT internalization, and neurons match endocytosis mediators to the different demands for locally recycling SCSVs or replenishing distally synthesized LDCVs. PMID:23640057

  20. Peptide inhibitors of botulinum neurotoxin serotype A: design, inhibition, cocrystal structures, structure-activity relationship and pharmacophore modeling

    Energy Technology Data Exchange (ETDEWEB)

    Kumar G.; Swaminathan S.; Kumaran, D.; Ahmed, S. A.

    2012-05-01

    Clostridium botulinum neurotoxins are classified as Category A bioterrorism agents by the Centers for Disease Control and Prevention (CDC). The seven serotypes (A-G) of the botulinum neurotoxin, the causative agent of the disease botulism, block neurotransmitter release by specifically cleaving one of the three SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins and induce flaccid paralysis. Using a structure-based drug-design approach, a number of peptide inhibitors were designed and their inhibitory activity against botulinum serotype A (BoNT/A) protease was determined. The most potent peptide, RRGF, inhibited BoNT/A protease with an IC{sub 50} of 0.9 {micro}M and a K{sub i} of 358 nM. High-resolution crystal structures of various peptide inhibitors in complex with the BoNT/A protease domain were also determined. Based on the inhibitory activities and the atomic interactions deduced from the cocrystal structures, the structure-activity relationship was analyzed and a pharmacophore model was developed. Unlike the currently available models, this pharmacophore model is based on a number of enzyme-inhibitor peptide cocrystal structures and improved the existing models significantly, incorporating new features.

  1. Comparative sequence analyses of the neurotoxin complex genes in Clostridium botulinum serotypes A, B, E, and F

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    Ajay K. Singh

    2012-09-01

    Full Text Available Neurotoxin complex (NTC genes are arranged in two known hemagglutinin (HA and open reading frame X (ORFX clusters. NTC genes have been analyzed in four serotypes A, B, E and F of Clostridium botulinum causing human botulism. Analysis of amino acid sequences of NT genes demonstrated significant differences among subtypes and four serotypes. Phylogram tree of NT genes reveals that serotypes A1 and B1 are much closer compared to serotype E1 and F1. However, non-toxic non-hemagglutinin (NTNH gene is highly conserved among four serotypes. Analysis of phylogram tree of NTNH gene reveals that serotypes A and F are more closely related compared to serotype B and E. Additionally, sequences of HAs and ORFX genes are very divergent but these genes are specific in subtypes and serotypes of Clostridium botulinum. Information derived from sequence analyses of NTC has direct implication in development of detection tools and therapeutic countermeasures for botulism.

  2. Analysis of genomic differences among Clostridium botulinum type A1 strains

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    Singh Bal

    2010-12-01

    Full Text Available Abstract Background 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 studies. Results Several strategies were deployed in this report. First, University of Massachusetts Dartmouth laboratory Hall strain (UMASS strain neurotoxin gene was amplified by PCR and sequenced; its sequence was aligned with the published ATCC 3502 Sanger Institute Hall strain and Allergan Hall strain neurotoxin gene regions. Sequence alignment showed that there was a synonymous single nucleotide polymorphism (SNP in the region encoding the heavy chain between Allergan strain and ATCC 3502 and UMASS strains. Second, comparative genomic hybridization (CGH demonstrated that the UMASS strain and a strain expected to be derived from ATCC 3502 in the Centers for Disease Control and Prevention (CDC laboratory (ATCC 3502* differed in gene content compared to the ATCC 3502 genome sequence published by the Sanger Institute. Third, alignment of the three sequenced C. botulinum type A1 strain genomes revealed the presence of four comparable blocks. Strains ATCC 3502 and ATCC 19397 share the same genome organization, while the organization of the blocks in strain Hall were switched. Lastly, PCR was designed to identify UMASS and ATCC 3502* strain genome organizations. The PCR results indicated that UMASS strain belonged to Hall type and ATCC 3502* strain was identical to ATCC 3502 (Sanger Institute type. Conclusions Taken together, C. botulinum type A1 strains including Sanger Institute ATCC 3502, ATCC 3502*, ATCC 19397, Hall, Allergan, and

  3. Monoclonal Antibodies that Inhibit the Proteolytic Activity of Botulinum Neurotoxin Serotype/B

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    Yongfeng Fan

    2015-08-01

    Full Text Available Existing antibodies (Abs used to treat botulism cannot enter the cytosol of neurons and bind to botulinum neurotoxin (BoNT at its site of action, and thus cannot reverse paralysis. However, Abs targeting the proteolytic domain of the toxin could inhibit the proteolytic activity of the toxin intracellularly and potentially reverse intoxication, if they could be delivered intracellularly. As such, antibodies that neutralize toxin activity could serve as potent inhibitory cargos for therapeutic antitoxins against botulism. BoNT serotype B (BoNT/B contains a zinc endopeptidase light chain (LC domain that cleaves synaoptobrevin-2, a SNARE protein responsible for vesicle fusion and acetylcholine vesicle release. To generate monoclonal Abs (mAbs that could reverse paralysis, we targeted the protease domain for Ab generation. Single-chain variable fragment (scFv libraries from immunized mice or humans were displayed on yeast, and 19 unique BoNT/B LC-specific mAbs isolated by fluorescence-activated cell sorting (FACS. The equilibrium dissociation constants (KD of these mAbs for BoNT/B LC ranged from 0.24 nM to 14.3 nM (mean KD 3.27 nM. Eleven mAbs inhibited BoNT/B LC proteolytic activity. The fine epitopes of selected mAbs were identified by alanine-scanning mutagenesis, revealing that inhibitory mAbs bound near the active site, substrate-binding site or the extended substrate-binding site. The results provide mAbs that could prove useful for intracellular reversal of paralysis and identify epitopes that could be targeted by small molecules inhibitors.

  4. Monoclonal Antibodies Targeting the Alpha-Exosite of Botulinum Neurotoxin Serotype/A Inhibit Catalytic Activity.

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    Yongfeng Fan

    Full Text Available The paralytic disease botulism is caused by botulinum neurotoxins (BoNT, multi-domain proteins containing a zinc endopeptidase that cleaves the cognate SNARE protein, thereby blocking acetylcholine neurotransmitter release. Antitoxins currently used to treat botulism neutralize circulating BoNT but cannot enter, bind to or neutralize BoNT that has already entered the neuron. The light chain endopeptidase domain (LC of BoNT serotype A (BoNT/A was targeted for generation of monoclonal antibodies (mAbs that could reverse paralysis resulting from intoxication by BoNT/A. Single-chain variable fragment (scFv libraries from immunized humans and mice were displayed on the surface of yeast, and 19 BoNT/A LC-specific mAbs were isolated by using fluorescence-activated cell sorting (FACS. Affinities of the mAbs for BoNT/A LC ranged from a KD value of 9.0×10-11 M to 3.53×10-8 M (mean KD 5.38×10-9 M and median KD 1.53×10-9 M, as determined by flow cytometry analysis. Eleven mAbs inhibited BoNT/A LC catalytic activity with IC50 values ranging from 8.3 ~73×10-9 M. The fine epitopes of selected mAbs were also mapped by alanine-scanning mutagenesis, revealing that the inhibitory mAbs bound the α-exosite region remote from the BoNT/A LC catalytic center. The results provide mAbs that could prove useful for intracellular reversal of paralysis post-intoxication and further define epitopes that could be targeted by small molecule inhibitors.

  5. Toxic and nontoxic components of botulinum neurotoxin complex are evolved from a common ancestral zinc protein

    Energy Technology Data Exchange (ETDEWEB)

    Inui, Ken [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Japan Society for the Promotion of Science, 1-8 Chiyoda-ku, Tokyo 102-8472 (Japan); Sagane, Yoshimasa [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Miyata, Keita [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Japan Society for the Promotion of Science, 1-8 Chiyoda-ku, Tokyo 102-8472 (Japan); Miyashita, Shin-Ichiro [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Suzuki, Tomonori [Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 (Japan); Shikamori, Yasuyuki [Agilent Technologies International Japan, Ltd. Takaura-cho 9-1, Hachioji-shi, Tokyo 192-0033 (Japan); Ohyama, Tohru; Niwa, Koichi [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Watanabe, Toshihiro, E-mail: t-watana@bioindustry.nodai.ac.jp [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer BoNT and NTNHA proteins share a similar protein architecture. Black-Right-Pointing-Pointer NTNHA and BoNT were both identified as zinc-binding proteins. Black-Right-Pointing-Pointer NTNHA does not have a classical HEXXH zinc-coordinating motif similar to that found in all serotypes of BoNT. Black-Right-Pointing-Pointer Homology modeling implied probable key residues involved in zinc coordination. -- Abstract: Zinc atoms play an essential role in a number of enzymes. Botulinum neurotoxin (BoNT), the most potent toxin known in nature, is a zinc-dependent endopeptidase. Here we identify the nontoxic nonhemagglutinin (NTNHA), one of the BoNT-complex constituents, as a zinc-binding protein, along with BoNT. A protein structure classification database search indicated that BoNT and NTNHA share a similar domain architecture, comprising a zinc-dependent metalloproteinase-like, BoNT coiled-coil motif and concanavalin A-like domains. Inductively coupled plasma-mass spectrometry analysis demonstrated that every single NTNHA molecule contains a single zinc atom. This is the first demonstration of a zinc atom in this protein, as far as we know. However, the NTNHA molecule does not possess any known zinc-coordinating motif, whereas all BoNT serotypes possess the classical HEXXH motif. Homology modeling of the NTNHA structure implied that a consensus K-C-L-I-K-X{sub 35}-D sequence common among all NTNHA serotype molecules appears to coordinate a single zinc atom. These findings lead us to propose that NTNHA and BoNT may have evolved distinct functional specializations following their branching out from a common ancestral zinc protein.

  6. Enhanced neutralization potency of botulinum neurotoxin antibodies using a red blood cell-targeting fusion protein.

    Directory of Open Access Journals (Sweden)

    Sharad P Adekar

    Full Text Available Botulinum neurotoxin (BoNT potently inhibits cholinergic signaling at the neuromuscular junction. The ideal countermeasures for BoNT exposure are monoclonal antibodies or BoNT antisera, which form BoNT-containing immune complexes that are rapidly cleared from the general circulation. Clearance of opsonized toxins may involve complement receptor-mediated immunoadherence to red blood cells (RBC in primates or to platelets in rodents. Methods of enhancing immunoadherence of BoNT-specific antibodies may increase their potency in vivo. We designed a novel fusion protein (FP to link biotinylated molecules to glycophorin A (GPA on the RBC surface. The FP consists of an scFv specific for murine GPA fused to streptavidin. FP:mAb:BoNT complexes bound specifically to the RBC surface in vitro. In a mouse model of BoNT neutralization, the FP increased the potency of single and double antibody combinations in BoNT neutralization. A combination of two antibodies with the FP gave complete neutralization of 5,000 LD50 BoNT in mice. Neutralization in vivo was dependent on biotinylation of both antibodies and correlated with a reduction of plasma BoNT levels. In a post-exposure model of intoxication, FP:mAb complexes gave complete protection from a lethal BoNT/A1 dose when administered within 2 hours of toxin exposure. In a pre-exposure prophylaxis model, mice were fully protected for 72 hours following administration of the FP:mAb complex. These results demonstrate that RBC-targeted immunoadherence through the FP is a potent enhancer of BoNT neutralization by antibodies in vivo.

  7. Toxic and nontoxic components of botulinum neurotoxin complex are evolved from a common ancestral zinc protein

    International Nuclear Information System (INIS)

    Highlights: ► BoNT and NTNHA proteins share a similar protein architecture. ► NTNHA and BoNT were both identified as zinc-binding proteins. ► NTNHA does not have a classical HEXXH zinc-coordinating motif similar to that found in all serotypes of BoNT. ► Homology modeling implied probable key residues involved in zinc coordination. -- Abstract: Zinc atoms play an essential role in a number of enzymes. Botulinum neurotoxin (BoNT), the most potent toxin known in nature, is a zinc-dependent endopeptidase. Here we identify the nontoxic nonhemagglutinin (NTNHA), one of the BoNT-complex constituents, as a zinc-binding protein, along with BoNT. A protein structure classification database search indicated that BoNT and NTNHA share a similar domain architecture, comprising a zinc-dependent metalloproteinase-like, BoNT coiled-coil motif and concanavalin A-like domains. Inductively coupled plasma-mass spectrometry analysis demonstrated that every single NTNHA molecule contains a single zinc atom. This is the first demonstration of a zinc atom in this protein, as far as we know. However, the NTNHA molecule does not possess any known zinc-coordinating motif, whereas all BoNT serotypes possess the classical HEXXH motif. Homology modeling of the NTNHA structure implied that a consensus K-C-L-I-K-X35-D sequence common among all NTNHA serotype molecules appears to coordinate a single zinc atom. These findings lead us to propose that NTNHA and BoNT may have evolved distinct functional specializations following their branching out from a common ancestral zinc protein.

  8. Characterization and immunological activity of different forms of recombinant secreted Hc of botulinum neurotoxin serotype B products expressed in yeast.

    Science.gov (United States)

    Liu, Bo; Shi, DanYang; Chang, ShaoHong; Gong, Xin; Yu, YunZhou; Sun, ZhiWei; Wu, Jun

    2015-01-01

    The recombinant Hc proteins of botulinum neurotoxins and tetanus toxin are exclusively produced by intracellular heterologous expression in Pichia pastoris for use in subunit vaccines; the same Hc proteins produced by secreted heterologous expression are hyper-glycosylated and immunologically inert. Here, several different recombinant secreted Hc proteins of botulinum neurotoxin serotype B (BHc) were expressed in yeast and we characterized and assessed their immunological activity in detail. Recombinant low-glycosylated secreted BHc products (BSK) were also immunologically inert, similar to hyper-glycosylated BHc products (BSG), although deglycosylation restored their immunological activities. Unexpectedly, deglycosylated proBHc contained an unexpected pro-peptide of an α-factor signal and fortuitous N-linked glycosylation sites in the non-cleaved pro-peptide sequences, but not in the BHc sequences. Notably, a non-glycosylated secreted homogeneous BHc isoform (mBHc), which we successfully prepared after deleting the pro-peptide and removing its single potential glycosylation site, was immunologically active and could confer effective protective immunity, similarly to non-glycosylated rBHc. In summary, we conclude that a non-glycosylated secreted BHc isoform can be prepared in yeast by deleting the pro-peptide of the α-factor signal and mutating its single potential glycosylation site. This approach provides a rational and feasible strategy for the secretory expression of botulism or other toxin antigens. PMID:25567004

  9. The ultimate radiochemical nightmare : upon radio-iodination of Botulinum neurotoxin A, the introduced iodine atom itself seems to be fatal for the bioactivity of this macromolecule

    NARCIS (Netherlands)

    Uhm, J.I.M. van; Visser, G.W.M.; Schans, M.J. van der; Geldof, A.A.; Meuleman, E.J.H.; Nieuwenhuijzen, J.A.

    2015-01-01

    Background: Botulinum neurotoxin A (BoNT-A) is a highly neurotoxic drug and frequently used in patients. Knowledge on the optimal way of administration of BoNT-A and its subsequent distribution is still rather limited. An accurate method for monitoring these processes might be the use of radiolabell

  10. Safety and administration of treatment with botulinum neurotoxin for sialorrhoea in ALS patients: Review of the litterature and a proposal for tailored treatment

    DEFF Research Database (Denmark)

    Stokholm, Morten; Bisgård, Carsten; Vilholm, Ole Jakob

    2013-01-01

    Botulinum neurotoxin (BoNT) is a second-line treatment of sialorrhoea in ALS (amyotrophic lateral sclerosis) patients. This article is a review of the published literature concerning safety and administration of this treatment to ALS patients. A PubMed search was performed. All original...

  11. No Decrease in Muscle Strength after Botulinum Neurotoxin-A Injection in Children with Cerebral Palsy

    Science.gov (United States)

    Eek, Meta N.; Himmelmann, Kate

    2016-01-01

    Spasticity and muscle weakness is common in children with cerebral palsy (CP). Spasticity can be treated with botulinum neurotoxin-A (BoNT-A), but this drug has also been reported to induce muscle weakness. Our purpose was to describe the effect on muscle strength in the lower extremities after BoNT-A injections in children with CP. A secondary aim was to relate the effect of BoNT-A to gait pattern and range of motion. Twenty children with spastic CP were included in the study, 8 girls and 12 boys (mean age 7.7 years). All were able to walk without support, but with increased muscle tone interfering with motor function and gait pattern. Sixteen children had unilateral spastic CP and four bilateral spastic CP. Twenty-four legs received injections with BoNT-A in the plantar flexor muscles. The children were tested before treatment, around 6 weeks after at the peak effect of BoNT-A, and at 6 months after treatment, with measurement of muscle strength, gait analysis, and range of motion. There were no differences in muscle strength in plantar flexors of treated legs at peak effect compared to baseline. Six months after treatment, there was still no change in untreated plantar flexor muscles, but an increasing trend in plantar flexor strength in legs treated with BoNT-A. Parents reported positive effects in all children, graded as: small in three children, moderate in eight, and large in nine children. The gait analysis showed a small improvement in knee extension at initial contact, and there was a small increase in passive range of motion for ankle dorsiflexion. Two children had a period with transient weakness and pain. We found that voluntary force production in plantar flexor muscles did not decrease after BoNT-A, instead there was a trend to increased muscle strength at follow-up. The increase may be explained as an effect of the blocking of involuntary nerve impulses, leading to an opportunity to using and training the muscles with voluntary control. Adequate

  12. Glycosylated SV2 and Gangliosides as Dual Receptors for Botulinum Neurotoxin Serotype F

    Energy Technology Data Exchange (ETDEWEB)

    Fu, Zhuji; Chen, Chen; Barbieri, Joseph T.; Kim, Jung-Ja P.; Baldwin, Michael R.; (MCW)

    2010-02-22

    Botulinum neurotoxin causes rapid flaccid paralysis through the inhibition of acetylcholine release at the neuromuscular junction. The seven BoNT serotypes (A-G) have been proposed to bind motor neurons via ganglioside-protein dual receptors. To date, the structure-function properties of BoNT/F host receptor interactions have not been resolved. Here, we report the crystal structures of the receptor binding domains (HCR) of BoNT/A and BoNT/F and the characterization of the dual receptors for BoNT/F. The overall polypeptide fold of HCR/A is essentially identical to the receptor binding domain of the BoNT/A holotoxin, and the structure of HCR/F is very similar to that of HCR/A, except for two regions implicated in neuronal binding. Solid phase array analysis identified two HCR/F binding glycans: ganglioside GD1a and oligosaccharides containing an N-acetyllactosamine core. Using affinity chromatography, HCR/F bound native synaptic vesicle glycoproteins as part of a protein complex. Deglycosylation of glycoproteins using {alpha}(1-3,4)-fucosidase, endo-{beta}-galactosidase, and PNGase F disrupted the interaction with HCR/F, while the binding of HCR/B to its cognate receptor, synaptotagmin I, was unaffected. These data indicate that the HCR/F binds synaptic vesicle glycoproteins through the keratan sulfate moiety of SV2. The interaction of HCR/F with gangliosides was also investigated. HCR/F bound specifically to gangliosides that contain {alpha}2,3-linked sialic acid on the terminal galactose of a neutral saccharide core (binding order GT1b = GD1a GM3; no binding to GD1b and GM1a). Mutations within the putative ganglioside binding pocket of HCR/F decreased binding to gangliosides, synaptic vesicle protein complexes, and primary rat hippocampal neurons. Thus, BoNT/F neuronal discrimination involves the recognition of ganglioside and protein (glycosylated SV2) carbohydrate moieties, providing a structural basis for the high affinity and specificity of BoNT/F for neurons.

  13. Mass spectrometry-based methods for detection and differentiation of botulinum neurotoxins

    Science.gov (United States)

    Schmidt, Jurgen G.; Boyer, Anne E.; Kalb, Suzanne R.; Moura, Hercules; Barr, John R.; Woolfitt, Adrian R.

    2009-11-03

    The present invention is directed to a method for detecting the presence of clostridial neurotoxins in a sample by mixing a sample with a peptide that can serve as a substrate for proteolytic activity of a clostridial neurotoxin; and measuring for proteolytic activity of a clostridial neurotoxin by a mass spectroscopy technique. In one embodiment, the peptide can have an affinity tag attached at two or more sites.

  14. Botulinum toxin.

    OpenAIRE

    Savardekar Preeti

    1989-01-01

    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...

  15. Interneuronal Transfer and Distal Action of Tetanus Toxin and Botulinum Neurotoxins A and D in Central Neurons.

    Science.gov (United States)

    Bomba-Warczak, Ewa; Vevea, Jason D; Brittain, Joel M; Figueroa-Bernier, Annette; Tepp, William H; Johnson, Eric A; Yeh, Felix L; Chapman, Edwin R

    2016-08-16

    Recent reports suggest that botulinum neurotoxin (BoNT) A, which is widely used clinically to inhibit neurotransmission, can spread within networks of neurons to have distal effects, but this remains controversial. Moreover, it is not known whether other members of this toxin family are transferred between neurons. Here, we investigate the potential distal effects of BoNT/A, BoNT/D, and tetanus toxin (TeNT), using central neurons grown in microfluidic devices. Toxins acted upon the neurons that mediated initial entry, but all three toxins were also taken up, via an alternative pathway, into non-acidified organelles that mediated retrograde transport to the somato-dendritic compartment. Toxins were then released into the media, where they entered and exerted their effects upon upstream neurons. These findings directly demonstrate that these agents undergo transcytosis and interneuronal transfer in an active form, resulting in long-distance effects. PMID:27498860

  16. Detection of botulinum neurotoxin serotype B at sub mouse LD(50 levels by a sandwich immunoassay and its application to toxin detection in milk.

    Directory of Open Access Journals (Sweden)

    Miles C Scotcher

    Full Text Available BACKGROUND: 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. METHODS AND FINDINGS: Recombinant peptide fragments of the light chain, the transmembrane and receptor-binding domains of the heavy chain of botulinum neurotoxin type B (BoNT/B were expressed in Escherichia coli as GST-fusion proteins and purified. These proteins were used to immunize BALB/cJ mice for the generation of monoclonal antibodies (mAbs. Antibody-producing hybridomas were detected using either a direct binding ELISA binding to plate-immobilized BoNT/B, or with a capture-capture ELISA whereby the capacity of the antibody to capture BoNT/B from solution was tested. A total of five mAbs were selected, two of which bound the toxin light chain and three bound the receptor-binding domain of BoNT/B heavy chain. MAb MCS6-27 was identified via capture-capture ELISA and was the only mAb able to bind BoNT/B in solution under physiological conditions. MAbs F24-1, F26-16, F27-33 and F29-40 were identified via direct binding ELISA, and were able to capture BoNT/B in solution only in the presence of 0.5-0.9 mM sodium dodecyl sulphate (SDS. MAb MCS6-27 and an anti-BoNT/B polyclonal antibody were incorporated into a sandwich ELISA that did not require SDS. CONCLUSIONS: We report here the generation of monoclonal antibodies to serotype B and the subsequent development of a sensitive sandwich immunoassay. This immunoassay has a detection limit of 100 fg BoNT/B, fifty times more sensitive than the mouse bioassay detection limit of 5 pg BoNT/B. Additionally, this assay detected as little as 39 pg/mL of toxin in skim, 2% and whole milk.

  17. Clinical use of non-A botulinum toxins: botulinum toxin type B.

    Science.gov (United States)

    Dressler, D; Eleopra, R

    2006-04-01

    Botulinum neurotoxin type B (BT, BT-B) has been used as NeuroBloc/MyoBloc since 1999 for treatment of cervical dystonia, hyperhidrosis, spastic conditions, cerebral palsy, hemifacial spasm, bladder dysfunction, spasmodic dysphonia, sialorrhoea, anal fissures, piriformis syndrome, various pain conditions and cosmetic applications. Generally, its therapeutic effects are comparable to BT type A (BT-A). The adverse effect profiles of BT-B and BT-A, however, differ considerably. BT-B has been found to produce more regional as well as systemic anticholinergic adverse effects, such as dryness of mouth, accommodation difficulties, conjunctival irritation, reduced sweating, dysphagia, heartburn, constipation, bladder voiding difficulties and dryness of nasal mucosa. In BT-B the relationship between autonomic and motor effects known from BT-A is substantially shifted towards autonomic effects. BT-B, therefore, should be used carefully in patients with autonomic disorders and in patients with concomitant anticholinergic therapy. If NeuroBloc/MyoBloc is used to treat cervical dystonia patients with antibody-induced failure of BT-A therapy, 86% of those will develop complete secondary therapy failure after five applications. If NeuroBloc/MyoBloc used to treat cervical dystonia patients without prior exposure to BT, 44% of those will develop complete secondary therapy failure after nine applications. NeuroBloc/MyoBloc, therefore, is associated with substantial antigenicity problems originating from a particular low specific biological potency. Systemic anticholinergic adverse effects and high antigenicity limits the clinical use of NeuroBloc/MyoBloc considerably. PMID:16785108

  18. Botulinum and Tetanus Neurotoxin-Induced Blockade of Synaptic Transmission in Networked Cultures of Human and Rodent Neurons.

    Science.gov (United States)

    Beske, Phillip H; Bradford, Aaron B; Grynovicki, Justin O; Glotfelty, Elliot J; Hoffman, Katie M; Hubbard, Kyle S; Tuznik, Kaylie M; McNutt, Patrick M

    2016-02-01

    Clinical manifestations of tetanus and botulism result from an intricate series of interactions between clostridial neurotoxins (CNTs) and nerve terminal proteins that ultimately cause proteolytic cleavage of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins and functional blockade of neurotransmitter release. Although detection of cleaved SNARE proteins is routinely used as a molecular readout of CNT intoxication in cultured cells, impaired synaptic function is the pathophysiological basis of clinical disease. Work in our laboratory has suggested that the blockade of synaptic neurotransmission in networked neuron cultures offers a phenotypic readout of CNT intoxication that more closely replicates the functional endpoint of clinical disease. Here, we explore the value of measuring spontaneous neurotransmission frequencies as novel and functionally relevant readouts of CNT intoxication. The generalizability of this approach was confirmed in primary neuron cultures as well as human and mouse stem cell-derived neurons exposed to botulinum neurotoxin serotypes A-G and tetanus neurotoxin. The sensitivity and specificity of synaptic activity as a reporter of intoxication was evaluated in assays representing the principal clinical and research purposes of in vivo studies. Our findings confirm that synaptic activity offers a novel and functionally relevant readout for the in vitro characterizations of CNTs. They further suggest that the analysis of synaptic activity in neuronal cell cultures can serve as a surrogate for neuromuscular paralysis in the mouse lethal assay, and therefore is expected to significantly reduce the need for terminal animal use in toxin studies and facilitate identification of candidate therapeutics in cell-based screening assays. PMID:26615023

  19. Effect of Botulinum Neurotoxin A Injection into the Submucoperichondrium of the Nasal Septum in Reducing Idiopathic Non-Allergic Rhinitis and Persistent Allergic Rhinitis

    OpenAIRE

    Mozafarinia, Keramat; Abna, Mehdi; Narges KHANJANI

    2015-01-01

    Introduction: Submucoperichondrial injection of botulinum neurotoxin A (BTA) in the nasal septum is a promising therapeutic option in the treatment of persistent allergic rhinitis (AR) and non-allergic rhinitis, and is safer and more effective than intraturbinate injection in reducing clinical symptoms. Materials and Methods: Forty patients diagnosed with persistent AR or non-allergic rhinitis referred to Shafa Medical Center affiliated to Kerman University of Medical Sciences were included i...

  20. Acrylamide inhibits nerve sprouting induced by botulinum toxin type A

    Institute of Scientific and Technical Information of China (English)

    Hong Jiang; Yi Xiang; Xingyue Hu; Huaying Cai

    2014-01-01

    Botulinum toxin type A is a potent muscle relaxant that blocks the transmission and release of acetylcholine at the neuromuscular junction. Intramuscular injection of botulinum toxin type A has served as an effective and safe therapy for strabismus and focal dystonia. However, muscular weakness is temporary and after 3-4 months, muscle strength usually recovers because function-al recovery is mediated by nerve sprouting and reconstruction of the neuromuscular junction. Acrylamide may produce neurotoxic substances that cause retrograde necrotizing neuropathy and inhibit nerve sprouting caused by botulinum toxin type A. This study investigated whether acrylamide inhibits nerve sprouting after intramuscular injection of botulinum toxin type A. A tibial nerve sprouting model was established through local injection of botulinum toxin type A into the right gastrocnemius muscle of Sprague-Dawley rats. Following intramuscular injection, rats were given intraperitoneal injection of 3%acrylamide every 3 days for 21 days. Nerve sprout-ing appeared 2 weeks after intramuscular injection of botulinum toxin type A and single-fiber electromyography revealed abnormal conduction at the neuromuscular junction 1 week after intra-muscular injection of botulinum toxin type A. Following intraperitoneal injection of acrylamide, the peak muscle ifber density decreased. Electromyography jitter value were restored to normal levels 6 weeks after injection. This indicates that the maximal decrease in ifber density and the time at which functional conduction of neuromuscular junction was restored were delayed. Addition-ally, the increase in tibial nerve ifbers was reduced. Acrylamide inhibits nerve sprouting caused by botulinum toxin type A and may be used to prolong the clinical dosage of botulinum toxin type A.

  1. The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.

    Directory of Open Access Journals (Sweden)

    Sara Marinelli

    Full Text Available In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25, the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A, could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC. We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life.

  2. The Analgesic Effect on Neuropathic Pain of Retrogradely Transported botulinum Neurotoxin A Involves Schwann Cells and Astrocytes

    Science.gov (United States)

    Ricordy, Ruggero; Uggenti, Carolina; Tata, Ada Maria; Luvisetto, Siro; Pavone, Flaminia

    2012-01-01

    In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw’s nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients’ quality of life. PMID:23110146

  3. Attomolar detection of botulinum toxin type A in complex biological matrices.

    Directory of Open Access Journals (Sweden)

    Karine Bagramyan

    Full Text Available BACKGROUND: A highly sensitive, rapid and cost efficient method that can detect active botulinum neurotoxin (BoNT in complex biological samples such as foods or serum is desired in order to 1 counter the potential bioterrorist threat 2 enhance food safety 3 enable future pharmacokinetic studies in medical applications that utilize BoNTs. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a botulinum neurotoxin serotype A assay with a large immuno-sorbent surface area (BoNT/A ALISSA that captures a low number of toxin molecules and measures their intrinsic metalloprotease activity with a fluorogenic substrate. In direct comparison with the "gold standard" mouse bioassay, the ALISSA is four to five orders of magnitudes more sensitive and considerably faster. Our method reaches attomolar sensitivities in serum, milk, carrot juice, and in the diluent fluid used in the mouse assay. ALISSA has high specificity for the targeted type A toxin when tested against alternative proteases including other BoNT serotypes and trypsin, and it detects the holotoxin as well as the multi-protein complex form of BoNT/A. The assay was optimized for temperature, substrate concentration, size and volume proportions of the immuno-sorbent matrix, enrichment and reaction times. Finally, a kinetic model is presented that is consistent with the observed improvement in sensitivity. CONCLUSIONS/SIGNIFICANCE: The sensitivity, specificity, speed and simplicity of the BoNT ALISSA should make this method attractive for diagnostic, biodefense and pharmacological applications.

  4. Botulinum toxin type A for the management of glabellar rhytids

    OpenAIRE

    Tremaine, Anne Marie

    2010-01-01

    Anne Marie Tremaine, Jerry L McCulloughDepartment of Dermatology, University of California, Irvine, CA, USAAbstract: There is an increasing demand for minimally-invasive cosmetic procedures to arrest the aging process. Botulinum toxin type A injections are the most commonly used nonsurgical cosmetic procedures in the United States. There has been research spanning over two decades dedicated to safety, efficacy, dosing, and complications of botulinum toxin type A. There are now two Food and Dr...

  5. 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.

    Science.gov (United States)

    Peck, Michael W; Plowman, June; Aldus, Clare F; Wyatt, Gary M; Izurieta, Walter Penaloza; Stringer, Sandra C; Barker, Gary C

    2010-10-01

    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 materials and foods. A new method that combines a selective enrichment culture with multiplex PCR has been developed and validated to enumerate specifically the spores of nonproteolytic C. botulinum. Key features of this new method include the following: (i) it is specific for nonproteolytic C. botulinum (and does not detect proteolytic C. botulinum), (ii) the detection limit has been determined for each food tested (using carefully structured control samples), and (iii) a low detection limit has been achieved by the use of selective enrichment and large test samples. The method has been used to enumerate spores of nonproteolytic C. botulinum in 637 samples of 19 food materials included in pasta-based minimally heated refrigerated foods and in 7 complete foods. A total of 32 samples (5 egg pastas and 27 scallops) contained spores of nonproteolytic C. botulinum type B or F. The majority of samples contained <100 spores/kg, but one sample of scallops contained 444 spores/kg. Nonproteolytic C. botulinum type E was not detected. Importantly, for QMRA and FSO, the construction of probability distributions will enable the frequency of packs containing particular levels of contamination to be determined.

  6. Computer-aided lead optimization: improved small-molecule inhibitor of the zinc endopeptidase of botulinum neurotoxin serotype A.

    Directory of Open Access Journals (Sweden)

    Jing Tang

    Full Text Available Optimization of a serotype-selective, small-molecule inhibitor of botulinum neurotoxin serotype A (BoNTA endopeptidase is a formidable challenge because the enzyme-substrate interface is unusually large and the endopeptidase itself is a large, zinc-binding protein with a complex fold that is difficult to simulate computationally. We conducted multiple molecular dynamics simulations of the endopeptidase in complex with a previously described inhibitor (K(i (app of 7+/-2.4 microM using the cationic dummy atom approach. Based on our computational results, we hypothesized that introducing a hydroxyl group to the inhibitor could improve its potency. Synthesis and testing of the hydroxyl-containing analog as a BoNTA endopeptidase inhibitor showed a twofold improvement in inhibitory potency (K(i (app of 3.8+/-0.8 microM with a relatively small increase in molecular weight (16 Da. The results offer an improved template for further optimization of BoNTA endopeptidase inhibitors and demonstrate the effectiveness of the cationic dummy atom approach in the design and optimization of zinc protease inhibitors.

  7. Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test.

    Science.gov (United States)

    Worbs, Sylvia; Fiebig, Uwe; Zeleny, Reinhard; Schimmel, Heinz; Rummel, Andreas; Luginbühl, Werner; Dorner, Brigitte G

    2015-12-01

    In the framework of the EU project EQuATox, a first international proficiency test (PT) on the detection and quantification of botulinum neurotoxins (BoNT) was conducted. Sample materials included BoNT serotypes A, B and E spiked into buffer, milk, meat extract and serum. Different methods were applied by the participants combining different principles of detection, identification and quantification. Based on qualitative assays, 95% of all results reported were correct. Successful strategies for BoNT detection were based on a combination of complementary immunological, MS-based and functional methods or on suitable functional in vivo/in vitro approaches (mouse bioassay, hemidiaphragm assay and Endopep-MS assay). Quantification of BoNT/A, BoNT/B and BoNT/E was performed by 48% of participating laboratories. It turned out that precise quantification of BoNT was difficult, resulting in a substantial scatter of quantitative data. This was especially true for results obtained by the mouse bioassay which is currently considered as "gold standard" for BoNT detection. The results clearly demonstrate the urgent need for certified BoNT reference materials and the development of methods replacing animal testing. In this context, the BoNT PT provided the valuable information that both the Endopep-MS assay and the hemidiaphragm assay delivered quantitative results superior to the mouse bioassay. PMID:26703724

  8. Generation and Characterization of Six Recombinant Botulinum Neurotoxins as Reference Material to Serve in an International Proficiency Test.

    Science.gov (United States)

    Weisemann, Jasmin; Krez, Nadja; Fiebig, Uwe; Worbs, Sylvia; Skiba, Martin; Endermann, Tanja; Dorner, Martin B; Bergström, Tomas; Muñoz, Amalia; Zegers, Ingrid; Müller, Christian; Jenkinson, Stephen P; Avondet, Marc-Andre; Delbrassinne, Laurence; Denayer, Sarah; Zeleny, Reinhard; Schimmel, Heinz; Åstot, Crister; Dorner, Brigitte G; Rummel, Andreas

    2015-12-01

    The detection and identification of botulinum neurotoxins (BoNT) is complex due to the existence of seven serotypes, derived mosaic toxins and more than 40 subtypes. Expert laboratories currently use different technical approaches to detect, identify and quantify BoNT, but due to the lack of (certified) reference materials, analytical results can hardly be compared. In this study, the six BoNT/A1-F1 prototypes were successfully produced by recombinant techniques, facilitating handling, as well as improving purity, yield, reproducibility and biosafety. All six BoNTs were quantitatively nicked into active di-chain toxins linked by a disulfide bridge. The materials were thoroughly characterized with respect to purity, identity, protein concentration, catalytic and biological activities. For BoNT/A₁, B₁ and E₁, serotypes pathogenic to humans, the catalytic activity and the precise protein concentration were determined by Endopep-mass spectrometry and validated amino acid analysis, respectively. In addition, BoNT/A₁, B₁, E₁ and F₁ were successfully detected by immunological assays, unambiguously identified by mass spectrometric-based methods, and their specific activities were assigned by the mouse LD50 bioassay. The potencies of all six BoNT/A1-F1 were quantified by the ex vivo mouse phrenic nerve hemidiaphragm assay, allowing a direct comparison. In conclusion, highly pure recombinant BoNT reference materials were produced, thoroughly characterized and employed as spiking material in a worldwide BoNT proficiency test organized by the EQuATox consortium. PMID:26703728

  9. Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test

    Directory of Open Access Journals (Sweden)

    Sylvia Worbs

    2015-11-01

    Full Text Available In the framework of the EU project EQuATox, a first international proficiency test (PT on the detection and quantification of botulinum neurotoxins (BoNT was conducted. Sample materials included BoNT serotypes A, B and E spiked into buffer, milk, meat extract and serum. Different methods were applied by the participants combining different principles of detection, identification and quantification. Based on qualitative assays, 95% of all results reported were correct. Successful strategies for BoNT detection were based on a combination of complementary immunological, MS-based and functional methods or on suitable functional in vivo/in vitro approaches (mouse bioassay, hemidiaphragm assay and Endopep-MS assay. Quantification of BoNT/A, BoNT/B and BoNT/E was performed by 48% of participating laboratories. It turned out that precise quantification of BoNT was difficult, resulting in a substantial scatter of quantitative data. This was especially true for results obtained by the mouse bioassay which is currently considered as “gold standard” for BoNT detection. The results clearly demonstrate the urgent need for certified BoNT reference materials and the development of methods replacing animal testing. In this context, the BoNT PT provided the valuable information that both the Endopep-MS assay and the hemidiaphragm assay delivered quantitative results superior to the mouse bioassay.

  10. In vitro potency determination of botulinum neurotoxin B based on its receptor-binding and proteolytic characteristics.

    Science.gov (United States)

    Wild, Emina; Bonifas, Ursula; Klimek, Jolanta; Trösemeier, Jan-Hendrik; Krämer, Beate; Kegel, Birgit; Behrensdorf-Nicol, Heike A

    2016-08-01

    Botulinum neurotoxins (BoNTs) are the most potent toxins known. However, the paralytic effect caused by BoNT serotypes A and B is taken advantage of to treat different forms of dystonia and in cosmetic procedures. Due to the increasing areas of application, the demand for BoNTs A and B is rising steadily. Because of the high toxicity, it is mandatory to precisely determine the potency of every produced BoNT batch, which is usually accomplished by performing toxicity testing (LD50 test) in mice. Here we describe an alternative in vitro assay for the potency determination of the BoNT serotype B. In this assay, the toxin is first bound to its specific receptor molecules. After the proteolytic subunit of the toxin has been released and activated by chemical reduction, it is exposed to synaptobrevin, its substrate protein. Finally the proteolytic cleavage is quantified by an antibody-mediated detection of the neoepitope, reaching a detection limit below 0.1mouseLD50/ml. Thus, the assay, named BoNT/B binding and cleavage assay (BoNT/B BINACLE), takes into account the binding as well as the protease function of the toxin, thereby measuring its biological activity. PMID:27032463

  11. Botulinum neurotoxin: Progress in negating its neurotoxicity; and in extending its therapeutic utility via molecular engineering. MiniReview.

    Science.gov (United States)

    Kostrzewa, Richard M; Kostrzewa, Rose Anna; Kostrzewa, John P

    2015-10-01

    While the poisonous effects of botulinum neurotoxin (BoNT) have been recognized since antiquity, the overall actions and mechanisms of effects of BoNT have been elucidated primarily over the past several decades. The general utility of BoNT is described in the paper, but the focus is mainly on the approaches towards negating the toxic effects of BoNT, and on the projection of an engineered BoNT molecule serving as a Trojan Horse to deliver a therapeutic load for treatment of a host of medical disorders. The BoNT molecule is configured with a binding domain, a zinc-dependent protease with specificity primarily for vesicular proteins, and a translocation domain for delivery of the metalloprotease into the cytoplasm. The anti-toxin approaches for BoNT include the use of vaccines, antibodies, block of BoNT binding or translocation, inhibition of metalloprotease activity, impeded translocation of the protease/catalytic domain, and inhibition of the downstream Src signaling pathway. Projections of BoNT as a therapeutic include its targeting to non-cholinergic nerves, also targeting to non-neuronal cells for treatment of hypersecretory disorders (e.g., cystic fibrosis), and treatment of hormonal disorders (e.g., acromegaly). Still in the exploratory phase, there is the expectation of major advances in BoNT neuroprotective strategies and burgeoning utility of engineered BoNTs as therapeutics. PMID:26192475

  12. Botulinum Neurotoxin A for Parotid Enlargement in Cystic Fibrosis: The First Case Report.

    Science.gov (United States)

    El Khoury, Joseph; Habre, Samer; Nasr, Marwan; Hokayem, Nabil

    2016-09-01

    Cystic fibrosis (CF) is the most common lethal autosomal recessive genetic disease associated with exocrine gland dysfunction. Salivary gland involvement is a common finding. The literature on submaxillary gland involvement has failed to address the parotid gland and any specific treatment of salivary gland manifestations of CF. Treatment is mainly symptomatic, consisting of analgesics, gustatory stimulation, and massage. Salivary secretion has clearly been linked to parasympathetic and sympathetic signals through intracellular calcium release. CF alters salivary composition with increased calcium and phosphorus concentrations and causes histologic changes (duct enlargement, dilation of acini, and abnormal mucous plugs). This study investigated whether botulinum toxin injected into the parotid gland during an acute exacerbation of CF-associated salivary gland disease could alleviate pain and control future exacerbations.

  13. The Effect of Total Cumulative Dose, Number of Treatment Cycles, Interval between Injections, and Length of Treatment on the Frequency of Occurrence of Antibodies to Botulinum Toxin Type A in the Treatment of Muscle Spasticity

    Science.gov (United States)

    Bakheit, Abdel Magid O.; Liptrot, Anthea; Newton, Rachel; Pickett, Andrew M.

    2012-01-01

    A large cumulative dose of botulinum toxin type A (BoNT-A), frequent injections, a short interval between treatment cycles, and a long duration of treatment have all been suggested, but not confirmed, to be associated with a high incidence of neutralizing antibodies to the neurotoxin. The aim of this study was to investigate whether these…

  14. Generation and Characterization of Six Recombinant Botulinum Neurotoxins as Reference Material to Serve in an International Proficiency Test

    Directory of Open Access Journals (Sweden)

    Jasmin Weisemann

    2015-11-01

    Full Text Available The detection and identification of botulinum neurotoxins (BoNT is complex due to the existence of seven serotypes, derived mosaic toxins and more than 40 subtypes. Expert laboratories currently use different technical approaches to detect, identify and quantify BoNT, but due to the lack of (certified reference materials, analytical results can hardly be compared. In this study, the six BoNT/A1–F1 prototypes were successfully produced by recombinant techniques, facilitating handling, as well as improving purity, yield, reproducibility and biosafety. All six BoNTs were quantitatively nicked into active di-chain toxins linked by a disulfide bridge. The materials were thoroughly characterized with respect to purity, identity, protein concentration, catalytic and biological activities. For BoNT/A1, B1 and E1, serotypes pathogenic to humans, the catalytic activity and the precise protein concentration were determined by Endopep-mass spectrometry and validated amino acid analysis, respectively. In addition, BoNT/A1, B1, E1 and F1 were successfully detected by immunological assays, unambiguously identified by mass spectrometric-based methods, and their specific activities were assigned by the mouse LD50 bioassay. The potencies of all six BoNT/A1–F1 were quantified by the ex vivo mouse phrenic nerve hemidiaphragm assay, allowing a direct comparison. In conclusion, highly pure recombinant BoNT reference materials were produced, thoroughly characterized and employed as spiking material in a worldwide BoNT proficiency test organized by the EQuATox consortium.

  15. Intrastriatal injection of botulinum neurotoxin-A is not cytotoxic in rat brain - A histological and stereological analysis.

    Science.gov (United States)

    Mehlan, Juliane; Brosig, Hans; Schmitt, Oliver; Mix, Eilhard; Wree, Andreas; Hawlitschka, Alexander

    2016-01-01

    Parkinson's disease (PD) is caused by progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta, resulting in a deficiency of dopamine in the striatum and an increased release of acetylcholine by tonically active interneurons. Botulinum neurotoxin-A (BoNT-A) is well known for blocking transmitter release by cholinergic presynaptic terminals. Treating striatal hypercholinism by local application of BoNT-A could be a possible new local therapy option of PD. In previous studies of our group, we analyzed the effect of BoNT-A injection into the CPu of 6-OHDA lesioned hemiparkinsonian rats. Our studies showed that BoNT-A application in hemiparkinson rat model is capable of abolishing apomorphine induced rotations for approximately 3 months. Regularly occurring axonal swellings in the BoNT-A infiltrated striata were also discovered, which we named BoNT-A induced varicosities (BiVs). Résumé: Here we investigated the long-term effect of the injection of 1ng BoNT-A into the right CPu of naive Wistar rats on the number of ChAT-ir interneurons as well as on the numeric density and the volumetric size of the BiVs in the CPu. Significant differences in the number of ChAT-ir neurons between the right BoNT-A treated CPu and the left untreated CPu were not detected up to 12 month post BoNT-A injection. The numeric density of BiVs in the treated CPu reached a maximum 3 months after BoNT-A treatment and decreased afterwards, whereas the volume of single BiVs increased steadily throughout the whole time course of the experiment. PMID:26562665

  16. Accelerated intoxication of GABAergic synapses by botulinum neurotoxin A disinhibits stem cell-derived neuron networks prior to network silencing

    Directory of Open Access Journals (Sweden)

    Phillip H Beske

    2015-04-01

    Full Text Available Botulinum neurotoxins (BoNTs are extremely potent toxins that specifically cleave SNARE proteins in peripheral synapses, preventing neurotransmitter release. Neuronal responses to BoNT intoxication are traditionally studied by quantifying SNARE protein cleavage in vitro or monitoring physiological paralysis in vivo. Consequently, the dynamic effects of intoxication on synaptic behaviors are not well understood. We have reported that mouse embryonic stem cell-derived neurons (ESNs are highly sensitive to BoNT based on molecular readouts of intoxication. Here we study the time-dependent changes in synapse- and network-level behaviors following addition of BoNT/A to spontaneously active networks of glutamatergic and GABAergic ESNs. Whole-cell patch-clamp recordings indicated that BoNT/A rapidly blocked synaptic neurotransmission, confirming that ESNs replicate the functional pathophysiology responsible for clinical botulism. Quantitation of spontaneous neurotransmission in pharmacologically isolated synapses revealed accelerated silencing of GABAergic synapses compared to glutamatergic synapses, which was consistent with the selective accumulation of cleaved SNAP-25 at GAD1+ presynaptic terminals at early timepoints. Different latencies of intoxication resulted in complex network responses to BoNT/A addition, involving rapid disinhibition of stochastic firing followed by network silencing. Synaptic activity was found to be highly sensitive to SNAP-25 cleavage, reflecting the functional consequences of the localized cleavage of the small subpopulation of SNAP-25 that is engaged in neurotransmitter release in the nerve terminal. Collectively these findings illustrate that use of synaptic function assays in networked neurons cultures offers a novel and highly sensitive approach for mechanistic studies of toxin:neuron interactions and synaptic responses to BoNT.

  17. Reduced Neck Muscle Strength and Altered Muscle Mechanical Properties in Cervical Dystonia Following Botulinum Neurotoxin Injections: A Prospective Study

    Directory of Open Access Journals (Sweden)

    Sirpa Mustalampi

    2016-01-01

    Full Text Available Objective To evaluate changes in the strength and mechanical properties of neck muscles and disability in patients with cervical dystonia (CD during a 12-week period following botulinum neurotoxin (BoNT injections. Methods Eight patients with CD volunteered for this prospective clinical cohort study. Patients had received BoNT injections regularly in neck muscles at three-month intervals for several years. Maximal isometric neck strength was measured by a dynamometer, and the mechanical properties of the splenius capitis were evaluated using two myotonometers. Clinical assessment was performed using the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS before and at 2, 4, 8, and 12 weeks after the BoNT injections. Results Mean maximal isometric neck strength at two weeks after the BoNT injections decreased by 28% in extension, 25% in rotation of the affected side and 17% in flexion. At four weeks, muscle stiffness of the affected side decreased by 17% and tension decreased by 6%. At eight weeks, the muscle elasticity on the affected side increased by 12%. At two weeks after the BoNT injections, the TWSTRS-severity and TWSTRS-total scores decreased by 4.3 and 6.4, respectively. The strength, muscle mechanical properties and TWSTRS scores returned to baseline values at 12 weeks. Conclusions Although maximal neck strength and muscle tone decreased after BoNT injections, the disability improved. The changes observed after BoNT injections were temporary and returned to pre-injection levels within twelve weeks. Despite having a possible negative effect on function and decreasing neck strength, the BoNT injections improved the patients reported disability.

  18. Reduced Neck Muscle Strength and Altered Muscle Mechanical Properties in Cervical Dystonia Following Botulinum Neurotoxin Injections: A Prospective Study

    Science.gov (United States)

    Mustalampi, Sirpa; Ylinen, Jari; Korniloff, Katariina; Weir, Adam; Häkkinen, Arja

    2016-01-01

    Objective To evaluate changes in the strength and mechanical properties of neck muscles and disability in patients with cervical dystonia (CD) during a 12-week period following botulinum neurotoxin (BoNT) injections. Methods Eight patients with CD volunteered for this prospective clinical cohort study. Patients had received BoNT injections regularly in neck muscles at three-month intervals for several years. Maximal isometric neck strength was measured by a dynamometer, and the mechanical properties of the splenius capitis were evaluated using two myotonometers. Clinical assessment was performed using the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) before and at 2, 4, 8, and 12 weeks after the BoNT injections. Results Mean maximal isometric neck strength at two weeks after the BoNT injections decreased by 28% in extension, 25% in rotation of the affected side and 17% in flexion. At four weeks, muscle stiffness of the affected side decreased by 17% and tension decreased by 6%. At eight weeks, the muscle elasticity on the affected side increased by 12%. At two weeks after the BoNT injections, the TWSTRS-severity and TWSTRS-total scores decreased by 4.3 and 6.4, respectively. The strength, muscle mechanical properties and TWSTRS scores returned to baseline values at 12 weeks. Conclusions Although maximal neck strength and muscle tone decreased after BoNT injections, the disability improved. The changes observed after BoNT injections were temporary and returned to pre-injection levels within twelve weeks. Despite having a possible negative effect on function and decreasing neck strength, the BoNT injections improved the patients reported disability. PMID:26828215

  19. Catalytic Features of the Botulinum Neurotoxin A Light Chain Revealed by High Resolution Structure of an Inhibitory Peptide Complex

    Energy Technology Data Exchange (ETDEWEB)

    Silvaggi,N.; Wilson, D.; Tzipori, S.; Allen, K.

    2008-01-01

    The Clostridium botulinum neurotoxin serotype A light chain (BoNT/A-LC) is a Zn(II)-dependent metalloprotease that blocks the release of acetylcholine at the neuromuscular junction by cleaving SNAP-25, one of the SNARE proteins required for exocytosis. Because of the potential for use of the toxin in bioterrorism and the increasingly widespread application of the toxin in the medical field, there is significant interest in the development of small-molecule inhibitors of the metalloprotease. Efforts to design such inhibitors have not benefited from knowledge of how peptides bind to the active site since the enzyme-peptide structures available previously either were not occupied in the vicinity of the catalytic Zn(II) ion or did not represent the product of SNAP-25 substrate cleavage. Herein we report the 1.4 Angstroms-resolution X-ray crystal structure of a complex between the BoNT/A-LC and the inhibitory peptide N-Ac-CRATKML, the first structure of the light chain with an inhibitory peptide bound at the catalytic Zn(II) ion. The peptide is bound with the Cys S? atom coordinating the metal ion. Surprisingly, the cysteine sulfur is oxidized to the sulfenic acid form. Given the unstable nature of this species in solution, is it likely that oxidation occurs on the enzyme. In addition to the peptide-bound structure, we report two structures of the unliganded light chain with and without the Zn(II) cofactor bound at 1.25 and 1.20 Angstroms resolution, respectively. The two structures are nearly identical, confirming that the Zn(II) ion plays a purely catalytic role. Additionally, the structure of the Zn(II)-bound uncomplexed enzyme allows identification of the catalytic water molecule and a second water molecule that occupies the same position as the peptidic oxygen in the tetrahedral intermediate. This observation suggests that the enzyme active site is prearranged to stabilize the tetrahedral intermediate of the protease reaction.

  20. Spinal central effects of peripherally applied botulinum neurotoxin A in comparison between its subtypes A1 and A2

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    Hidetaka eKoizumi

    2014-06-01

    Full Text Available Because of its unique ability to exert long-lasting synaptic transmission blockade, botulinum neurotoxin A (BoNT/A is used to treat a wide variety of disorders involving peripheral nerve terminal hyperexcitability. However, it has been a matter of debate whether this toxin has central or peripheral sites of action. We employed a rat model in which BoNT/A1 or BoNT/A2 was unilaterally injected into the gastrocnemius muscle. On time-course measurements of compound muscle action potential (CMAP amplitudes after injection of BoNT/A1 or BoNT/A2 at doses ranging from 1.7 U to 13.6 U, CMAP amplitude for the ipsilateral hind leg was markedly decreased on the 1st day, and this muscle flaccidity persisted up to the 14th day. Of note, both BoNT/A1 and BoNT/A2 administrations also resulted in decreased CMAP amplitudes for the contralateral leg in a dose-dependent manner ranging from 1.7 to 13.6 U, and this muscle flaccidity increased until the 4th day and then slowly recovered. Immunohistochemical results revealed that BoNT/A-cleaved SNAP-25 (synaptosomal-associated protein of 25 kDa appeared in the bilateral ventral and dorsal horns 4 days after injection of BoNT/A1 (10 U or BoNT/A2 (10 U, although there seemed to be a wider spread of BoNT/A-cleaved SNAP-25 associated with BoNT/A1 than BoNT/A2 in the contralateral spinal cord. This suggests that the catalytically active BoNT/A1 and BoNT/A2 were axonally transported via peripheral motor and sensory nerves to the spinal cord, where they spread through a transcytosis (cell-to-cell trafficking mechanism. Our results provide evidence for the central effects of intramuscularly administered BoNT/A1 and BoNT/A2 in the spinal cord, and a new insight into the clinical effects of peripheral BoNT/A applications.

  1. Investigations into an Outbreak of Botulism Caused by Clostridium botulinum Type C/D in Laying Hens.

    Science.gov (United States)

    Skarin, Hanna; Lindgren, Ylva; Jansson, Désirée S

    2015-06-01

    This case report describes a recent botulism outbreak in commercial laying hens with a history of increased mortality and flaccid paralysis. Routine diagnostic gross examination and microscopy from seven hens were inconclusive, but botulinum neurotoxin (BoNT) in peripheral blood was neutralized with both type C and type D antitoxins in the mouse bioassay. During a farm visit, 10 additional hens from a 34-wk-old flock on the farm were selected for clinical examination and further sampling. Nine hens were observed in sternal recumbency, with flaccid paralysis of the neck, drooping wings and tail, inability to escape, and bilateral ptosis, and one hen showed nonspecific clinical signs. Samples from cecum and liver were collected, and the gene coding for BoNT was detected by PCR in all 10 cecal samples and in four of the liver samples. Clostridium botulinum mosaic type C/D was isolated from 5 out of 10 hens from either cecum or liver, and the isolates were subjected to pulsed-field gel electrophoresis subtyping. All five isolates produced the same banding pattern, which was identical or showed >90% similarity to isolates from three different outbreaks on broiler farms in Sweden and Denmark during the 2007-10 period. However, they were clearly distinguishable from the predominantly reported pulsotype associated with avian botulism outbreaks in Europe. The authors are unaware of any previous report of C. botulinum mosaic type C/D isolates from laying hens.

  2. Detection of Clostridium botulinum neurotoxin genes (A-F) in dairy farms from Northern Germany using PCR: A case-control study.

    Science.gov (United States)

    Fohler, Svenja; Discher, Sabrina; Jordan, Eva; Seyboldt, Christian; Klein, Guenter; Neubauer, Heinrich; Hoedemaker, Martina; Scheu, Theresa; Campe, Amely; Charlotte Jensen, Katharina; Abdulmawjood, Amir

    2016-06-01

    Classical botulism in cattle mainly occurs after ingestion of feed contaminated with preformed toxin. In 2001 a form of botulism ("visceral botulism") was postulated to occur after ingestion of Clostridium (C.) botulinum cells or spores, followed by colonization of the intestine, and local production of botulinum neurotoxin (BoNT) causing chronic generalized disease. To verify the potential role of C. botulinum in the described syndrome, a case-control study was conducted, including 139 farms. Fecal samples, rumen content, water and silage samples were collected on each farm. Real time BoNT gene PCR assays were conducted after enrichment in RCM (Reinforced Clostridial Medium) at 37 °C and conventional PCRs after enrichment in MCM (Modified Cooked Meat Medium) at 30 °C. Furthermore, a direct detection of BoNT genes without prior enrichment was attempted. BoNT A, B, C, D, E and F genes were detected in animal samples from 25 (17.99%), 3 (2.16%), 0 (0.0%), 2 (1.44%), 1 (0.72%), and 3 (2.16%) farms, respectively. Eleven feed samples were positive for BoNT A gene. By enrichment a significant increase in sensitivity was achieved. Therefore, this should be an essential part of any protocol. No significant differences regarding BoNT gene occurrence could be observed between Case and Control farms or chronically diseased and clinically healthy animals within the particular category. Thus, the postulated form of chronic botulism in cows could not be confirmed. This study supports the general opinion that C. botulinum can occasionally be found in the rumen and intestine of cows without causing disease. PMID:27016061

  3. Easy expression of the C-terminal heavy chain domain of botulinum neurotoxin serotype A as a vaccine candidate using a bi-cistronic baculovirus system.

    Science.gov (United States)

    Villaflores, Oliver B; Hsei, Chein-Ming; Teng, Chao-Yi; Chen, Ying-Ju; Wey, Jiunn-Jye; Tsui, Pei-Yi; Shyu, Rong-Hwa; Tung, Kuo-Lun; Yeh, Jui-Ming; Chiao, Der-Jiang; Wu, Tzong-Yuan

    2013-04-01

    Clostridial botulinum neurotoxin (BoNT) is one of the most toxic proteins causing the food borne disease, botulism. In previous studies, recombinant BoNT production by Escherichia coli and yeast Pichia pastoris has been hampered by high AT content and codon bias in the gene encoding BoNT and required a synthetic gene to resolve this intrinsic bottleneck. This paper reports the simultaneous expression of the C-terminal heavy chain domain of BoNT (rBoNT/A-HC-6h) and enhanced green fluorescent protein (EGFP) using a bi-cistronic baculovirus-insect cell expression system. The expression of EGFP facilitated the monitoring of viral infection, virus titer determination, and isolation of the recombinant virus. Protein fusion with hexa-His-tag and one-step immobilized metal-ion affinity chromatography (IMAC) purification produced a homogenous, stable, and immunologically active 55-kDa rBoNT/A-HC-6h (about 3mg/L) with >90% purity. Furthermore, measured levels of serum titers were 8-folds for mice vaccinated with the purified rBoNT/A-HC-6h (2μg) than for mice administered with botulinum toxoid after initial immunization. Challenge experiment with botulinum A toxin demonstrated the immunoprotective activity of purified rBoNT/A-HC-6h providing the mice full protection against 10(2) LD50 botulinum A toxin with a dose as low as 0.2μg. This study provided supportive evidence for the use of a bi-cistronic baculovirus-Sf21 insect cell expression system in the facile expression of an immunogenically active rBoNT/A-HC. PMID:23313783

  4. Enzyme-amplified protein micorarray and a fluidic renewable surface fluorescence immunoassay for botulinum neurotoxin detection using high-affinity recombinant antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Varnum, Susan M.; Warner, Marvin G.; Dockendorff, Brian P.; Anheier, Norman C.; Lou, Jianlong; Marks, James D.; Smith, Leonard A.; Feldhaus, Michael J.; Grate, Jay W.; Bruckner-Lea, Cindy J.

    2006-06-16

    With the use of high-affinity recombinant monoclonal antibodies against the receptor binding domain of botulinum neurotoxin A (BoNT/A), two separate immunoassay platforms were developed for either the sensitive or the rapid detection of BoNT/A. An enzyme-linked immunosorbent assay (ELISA) microarray was developed for the specific and sensitive detection of BoNT in buffer and clinical fluids. This assay has the sensitivity to detect BoNT in diverse samples down to 14 fM (1.4 pg/mL). Using the recombinant monoclonal antibodies, a renewable surface microcolumn sensor was developed for the rapid detection of BoNT/A in an automated fluidic system. While the ELISA microarray assay, because of its sensitivity, offers an alternative to the mouse bioassay, the renewable surface assay has potential as a rapid screening assay for the analysis of complex environmental samples.

  5. Botulinum Toxin Type A as a Therapeutic Agent against Headache and Related Disorders

    OpenAIRE

    Siro Luvisetto; Parisa Gazerani; Carlo Cianchetti; Flaminia Pavone

    2015-01-01

    Botulinum neurotoxin A (BoNT/A) is a toxin produced by the naturally-occurring Clostridium botulinum that causes botulism. The potential of BoNT/A as a useful medical intervention was discovered by scientists developing a vaccine to protect against botulism. They found that, when injected into a muscle, BoNT/A causes a flaccid paralysis. Following this discovery, BoNT/A has been used for many years in the treatment of conditions of pathological muscle hyperactivity, like dystonias and spastic...

  6. 76 FR 29752 - Nomination of In Vitro Test Methods for Detection and Quantification of Botulinum Neurotoxins and...

    Science.gov (United States)

    2011-05-23

    ... meeting (67 FR 23323), comments and data are ] requested by June 2, 2011. NICEATM and ICCVAM will accept... HUMAN SERVICES Nomination of In Vitro Test Methods for Detection and Quantification of Botulinum... public comment on nominations received for (1) Three in vitro test methods proposed for detecting...

  7. Botulinum toxin

    Directory of Open Access Journals (Sweden)

    Nigam P

    2010-01-01

    Full Text Available 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 three months. Botulinum toxins now play a very significant role in the management of a wide variety of medical conditions, especially strabismus and focal dystonias, hemifacial spasm, and various spastic movement disorders, headaches, hypersalivation, hyperhidrosis, and some chronic conditions that respond only partially to medical treatment. The list of possible new indications is rapidly expanding. The cosmetological applications include correction of lines, creases and wrinkling all over the face, chin, neck, and chest to dermatological applications such as hyperhidrosis. Injections with botulinum toxin are generally well tolerated and side effects are few. A precise knowledge and understanding of the functional anatomy of the mimetic muscles is absolutely necessary to correctly use botulinum toxins in clinical practice.

  8. Brain Metabolic Changes of Cervical Dystonia with Spinocerebellar Ataxia Type 1 after Botulinum Toxin Therapy.

    Science.gov (United States)

    Kikuchi, Akio; Takeda, Atsushi; Sugeno, Naoto; Miura, Emiko; Kato, Kazuhiro; Hasegawa, Takafumi; Baba, Toru; Konno, Masatoshi; Oshima, Ryuji; Watanuki, Shoichi; Hiraoka, Kotaro; Tashiro, Manabu; Aoki, Masashi

    2016-01-01

    We occasionally observe long-term remission of cervical dystonia after several botulinum toxin treatments. However, botulinum toxin transiently acts on neuromuscular junctions. We herein report that a cervical dystonia patient with spinocerebellar ataxia type 1 could have long-term remission as a result of the depression of hypermetabolism in the bilateral putamen and primary sensorimotor cortex after botulinum toxin therapy. We suggest that botulinum toxin impacts the central nervous system, causing prolonged improvement through the normalization of basal ganglia circuits in addition to its effects at neuromuscular junctions. PMID:27432104

  9. A cross-sectional structured survey of patients receiving botulinum toxin type A treatment for blepharospasm.

    Science.gov (United States)

    Fezza, John; Burns, John; Woodward, Julie; Truong, Daniel; Hedges, Thomas; Verma, Amit

    2016-08-15

    To characterize satisfaction with current standard-of-care botulinum neurotoxin type A (BoNT/A) treatment for blepharospasm, we performed a cross-sectional, structured survey in subjects with blepharospasm who had received ≥2 BoNT/A cycles. Subjects were interviewed immediately before re-injection to evaluate treatment satisfaction, time course of treatment effects, preferred injection intervals, Jankovic Rating Scale (JRS), and Blepharospasm Disability Index (BSDI). Subjects' (n=114) last treatment was onabotulinumtoxinA (n=78), incobotulinumtoxinA (n=35), or abobotulinumtoxinA (n=1). The most frequent injection interval was 12weeks (46.5% subjects); 30.7% had an interval >12weeks. The main rationale for interval choice was "to maintain treatment efficacy" (44.7%). However, 36.6% reported that treatment effects usually declined within 8weeks; 69.6% within 10weeks. JRS and BSDI scores indicated re-emergence of symptoms before re-injection, with 70.2% and 73.7% of subjects reporting difficulties to drive and read, respectively. Overall, treatment satisfaction was high, but declined at the end of the cycle. Many subjects (52.3%) would prefer an injection interval of <12weeks; 30.6% of <10weeks. In conclusion, the survey results indicate that blepharospasm symptoms, such as difficulties to drive and read, re-emerge at the end of a BoNT treatment cycle and that flexible, individualized treatment intervals may improve treatment satisfaction and outcomes. PMID:27423565

  10. Efficacy and Safety of a New Botulinum Toxin Type A Free of Complexing Proteins

    Directory of Open Access Journals (Sweden)

    Hyun-Mi Oh

    2015-12-01

    Full Text Available MT10107 is botulinum neurotoxin type A derived drug which utilizes the 150 kDa portion without complexing proteins and human serum albumin contents. To evaluate the efficacy and the safety of MT10107, it was compared with onabotulinumtoxinA in this double-blind, randomized controlled trial. Twenty-five healthy males received a randomly selected dose of MT10107 into the extensor digitorum brevis (EDB muscle of one foot, and an equivalent dose of onabotulinumtoxinA (BOTOX was injected into the contralateral EDB muscle. While efficacy of the administered substance was determined by measuring paretic effects on the EDB, the local spread of toxin effects was evaluated by the paretic effects on the nearby abductor hallucis (AH and abductor digiti quinti (ADQ muscles. Paretic effects were defined as the percentage of reduction of the compound muscle action potential (CMAP amplitudes, measured at 14, 30, 90 days after the injection, compared to the baseline value. Intergroup (MT10107 and onabotulinumtoxinA differences were not significant in the percentage reduction of the amplitudes in the EDB muscles. In this study, there was no significant difference in efficacy and safety between the two test drugs. MT10107 may be effective and safe as much as onabotulinumtoxinA to produce the desired paretic effect.

  11. Efficacy and Safety of a New Botulinum Toxin Type A Free of Complexing Proteins.

    Science.gov (United States)

    Oh, Hyun-Mi; Park, Joo Hyun; Song, Dae Heon; Chung, Myung Eun

    2016-01-01

    MT10107 is botulinum neurotoxin type A derived drug which utilizes the 150 kDa portion without complexing proteins and human serum albumin contents. To evaluate the efficacy and the safety of MT10107, it was compared with onabotulinumtoxinA in this double-blind, randomized controlled trial. Twenty-five healthy males received a randomly selected dose of MT10107 into the extensor digitorum brevis (EDB) muscle of one foot, and an equivalent dose of onabotulinumtoxinA (BOTOX) was injected into the contralateral EDB muscle. While efficacy of the administered substance was determined by measuring paretic effects on the EDB, the local spread of toxin effects was evaluated by the paretic effects on the nearby abductor hallucis (AH) and abductor digiti quinti (ADQ) muscles. Paretic effects were defined as the percentage of reduction of the compound muscle action potential (CMAP) amplitudes, measured at 14, 30, 90 days after the injection, compared to the baseline value. Intergroup (MT10107 and onabotulinumtoxinA) differences were not significant in the percentage reduction of the amplitudes in the EDB muscles. In this study, there was no significant difference in efficacy and safety between the two test drugs. MT10107 may be effective and safe as much as onabotulinumtoxinA to produce the desired paretic effect. PMID:26712786

  12. Preferential entry of botulinum neurotoxin A Hc domain through intestinal crypt cells and targeting to cholinergic neurons of the mouse intestine.

    Directory of Open Access Journals (Sweden)

    Aurélie Couesnon

    Full Text Available Botulism, characterized by flaccid paralysis, commonly results from botulinum neurotoxin (BoNT absorption across the epithelial barrier from the digestive tract and then dissemination through the blood circulation to target autonomic and motor nerve terminals. The trafficking pathway of BoNT/A passage through the intestinal barrier is not yet fully understood. We report that intralumenal administration of purified BoNT/A into mouse ileum segment impaired spontaneous muscle contractions and abolished the smooth muscle contractions evoked by electric field stimulation. Entry of BoNT/A into the mouse upper small intestine was monitored with fluorescent HcA (half C-terminal domain of heavy chain which interacts with cell surface receptor(s. We show that HcA preferentially recognizes a subset of neuroendocrine intestinal crypt cells, which probably represent the entry site of the toxin through the intestinal barrier, then targets specific neurons in the submucosa and later (90-120 min in the musculosa. HcA mainly binds to certain cholinergic neurons of both submucosal and myenteric plexuses, but also recognizes, although to a lower extent, other neuronal cells including glutamatergic and serotoninergic neurons in the submucosa. Intestinal cholinergic neuron targeting by HcA could account for the inhibition of intestinal peristaltism and secretion observed in botulism, but the consequences of the targeting to non-cholinergic neurons remains to be determined.

  13. Identification of fibroblast growth factor receptor 3 (FGFR3 as a protein receptor for botulinum neurotoxin serotype A (BoNT/A.

    Directory of Open Access Journals (Sweden)

    Birgitte P S Jacky

    Full Text Available Botulinum neurotoxin serotype A (BoNT/A causes transient muscle paralysis by entering motor nerve terminals (MNTs where it cleaves the SNARE protein Synaptosomal-associated protein 25 (SNAP25206 to yield SNAP25197. Cleavage of SNAP25 results in blockage of synaptic vesicle fusion and inhibition of the release of acetylcholine. The specific uptake of BoNT/A into pre-synaptic nerve terminals is a tightly controlled multistep process, involving a combination of high and low affinity receptors. Interestingly, the C-terminal binding domain region of BoNT/A, HC/A, is homologous to fibroblast growth factors (FGFs, making it a possible ligand for Fibroblast Growth Factor Receptors (FGFRs. Here we present data supporting the identification of Fibroblast Growth Factor Receptor 3 (FGFR3 as a high affinity receptor for BoNT/A in neuronal cells. HC/A binds with high affinity to the two extra-cellular loops of FGFR3 and acts similar to an agonist ligand for FGFR3, resulting in phosphorylation of the receptor. Native ligands for FGFR3; FGF1, FGF2, and FGF9 compete for binding to FGFR3 and block BoNT/A cellular uptake. These findings show that FGFR3 plays a pivotal role in the specific uptake of BoNT/A across the cell membrane being part of a larger receptor complex involving ganglioside- and protein-protein interactions.

  14. High-level expression, purification, crystallization and preliminary X-ray crystallographic studies of the receptor binding domain of botulinum neurotoxin serotype D

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yanfeng; Gao, Xiaoli; Qin, Lin; Buchko, Garry W.; Robinson, Howard; Varnum, Susan M.

    2010-12-01

    Botulinum neurotoxins (BoNTs) are highly toxic proteins for humans and can cause neuroparalytic disease botulism. Due to the limitations of production and manipulation of holoenzymes, expressing non-toxic heavy chain receptor binding domains (HCR) has become a common strategy for vaccine and antibody development. Meanwhile, large quantities and highly purified soluble proteins are required for research areas such as antibody maturation and structural biology. We present high level expression and purification of the BoNT serotype D HCR in E. coli using a codon-optimized cDNA. By varying expression conditions, especially at low temperature, the protein was expressed at a high level with high solubility. About 150-200 mg protein was purified to >90% purity from 1 L cell culture. The recombinant D_HCR was crystallized and the crystals diffracted to 1.65 Å resolution. The crystals belong to space group P212121 with unit cell dimensions a = 60.8 Å, b = 89.7 Å, c = 93.9 Å. Preliminary crystallographic data analysis revealed one molecule in asymmetric unit.

  15. Botulinum Neurotoxin A Injections Influence Stretching of the Gastrocnemius Muscle-Tendon Unit in an Animal Model

    Directory of Open Access Journals (Sweden)

    Christopher J. Tuohy

    2012-08-01

    Full Text Available Botulinum Neurotoxin A (BoNT-A injections have been used for the treatment of muscle contractures and spasticity. This study assessed the influence of (BoNT-A injections on passive biomechanical properties of the muscle-tendon unit. Mouse gastrocnemius muscle (GC was injected with BoNT-A (n = 18 or normal saline (n = 18 and passive, non-destructive, in vivo load relaxation experimentation was performed to examine how the muscle-tendon unit behaves after chemical denervation with BoNT-A. Injection of BoNT-A impaired passive muscle recovery (15% vs. 35% recovery to pre-stretching baseline, p < 0.05 and decreased GC stiffness (0.531 ± 0.061 N/mm vs. 0.780 ± 0.037 N/mm, p < 0.05 compared to saline controls. The successful use of BoNT-A injections as an adjunct to physical therapy may be in part attributed to the disruption of the stretch reflex; thereby modulating in vivo passive muscle properties. However, it is also possible that BoNT-A injection may alter the structure of skeletal muscle; thus modulating the in vivo passive biomechanical properties of the muscle-tendon unit.

  16. Effect of Fill Temperature on Clostridium botulinum Type A Toxin Activity during the Hot Filling of Juice Bottles.

    Science.gov (United States)

    Skinner, Guy E; Fleischman, Gregory J; Balster, Fran; Reineke, Karl; Reddy, N Rukma; Larkin, John W

    2015-08-01

    The potential threat of terrorist attacks against the United States food supply using neurotoxin produced by Clostridium botulinum (BoNT) has resulted in the need for studying the effect of various food process operations on the bioavailability of this toxin. The objective of this study was to evaluate C. botulinum type A neurotoxin bioavailability after a simulated hot fill juice bottling operation. C. botulinum type A acid mud toxin (∼10(6) mouse lethal dose [MLD50]/ml) was deposited into juice bottles at an experimentally determined fastest cooling spot. Bottles (12 or 20 oz [355 and 592 ml]) were filled with either apple juice or an orange drink, at 80 or 85°C, in either upright or inverted orientations. Toxicity of the juice was evaluated as a function of holding time (1 to 2 min) by the mouse bioassay. The fastest cooling point in the upright orientation was determined to be at a bottle's bottom rim. In the inverted orientation, the fastest cooling point was in the bottle cap region. With respect to these two points, the upright bottle cooled faster than the inverted bottle, which was reflected in a higher inactivation of BoNT in the latter. For the orange drink (pH 2.9) toxicity was reduced by 0.5 × 10(6) MLD50/ml to a nondetectable level after 1 min in all bottle sizes, orientations, and temperatures as measured by the mouse bioassay. This indicates that there was at least a 0.5 × 10(6) MLD50/ml reduction in activity. Inactivation in apple juice (pH 4.0), to the same degree as in the orange drink, was found only for the inverted orientation at 85°C. Complete inactivation in apple juice for all conditions was found at a lower added toxin level of 0.25 × 10(5) MLD50/ml. In general, bottle inversion and filling at 85°C provided complete inactivation of BoNT to the 0.5 × 10(6) MLD50/ml level. All experiments resulted in the inactivation of 2.5 × 10(4) MLD50/ml of BoNT regardless of juice type, fill temperature, or bottle orientation and size.

  17. Effect of Fill Temperature on Clostridium botulinum Type A Toxin Activity during the Hot Filling of Juice Bottles.

    Science.gov (United States)

    Skinner, Guy E; Fleischman, Gregory J; Balster, Fran; Reineke, Karl; Reddy, N Rukma; Larkin, John W

    2015-08-01

    The potential threat of terrorist attacks against the United States food supply using neurotoxin produced by Clostridium botulinum (BoNT) has resulted in the need for studying the effect of various food process operations on the bioavailability of this toxin. The objective of this study was to evaluate C. botulinum type A neurotoxin bioavailability after a simulated hot fill juice bottling operation. C. botulinum type A acid mud toxin (∼10(6) mouse lethal dose [MLD50]/ml) was deposited into juice bottles at an experimentally determined fastest cooling spot. Bottles (12 or 20 oz [355 and 592 ml]) were filled with either apple juice or an orange drink, at 80 or 85°C, in either upright or inverted orientations. Toxicity of the juice was evaluated as a function of holding time (1 to 2 min) by the mouse bioassay. The fastest cooling point in the upright orientation was determined to be at a bottle's bottom rim. In the inverted orientation, the fastest cooling point was in the bottle cap region. With respect to these two points, the upright bottle cooled faster than the inverted bottle, which was reflected in a higher inactivation of BoNT in the latter. For the orange drink (pH 2.9) toxicity was reduced by 0.5 × 10(6) MLD50/ml to a nondetectable level after 1 min in all bottle sizes, orientations, and temperatures as measured by the mouse bioassay. This indicates that there was at least a 0.5 × 10(6) MLD50/ml reduction in activity. Inactivation in apple juice (pH 4.0), to the same degree as in the orange drink, was found only for the inverted orientation at 85°C. Complete inactivation in apple juice for all conditions was found at a lower added toxin level of 0.25 × 10(5) MLD50/ml. In general, bottle inversion and filling at 85°C provided complete inactivation of BoNT to the 0.5 × 10(6) MLD50/ml level. All experiments resulted in the inactivation of 2.5 × 10(4) MLD50/ml of BoNT regardless of juice type, fill temperature, or bottle orientation and size. PMID

  18. British Neurotoxin Network recommendations for managing cervical dystonia in patients with a poor response to botulinum toxin

    Science.gov (United States)

    Marion, Marie-Helene; Humberstone, Miles; Grunewald, Richard; Wimalaratna, Sunil

    2016-01-01

    Botulinum toxin (BoNT) injections are an effective treatment for cervical dystonia. Approximately 20% of patients eventually stop BoNT treatment, mostly because of treatment failure. These recommendations review the different therapeutic interventions for optimising the treatment in secondary poor responder patients. Immunoresistance has become less common over the years, but the diagnosis has to be addressed with a frontalis test or an Extensor Digitorum Brevis test. In case of immunoresistance to BoNT-A, we discuss the place the different therapeutic options (BoNT-A holidays, BoNT-B injections, alternative BoNT-A injections, deep brain stimulation). When poor responders are not immunoresistant, they benefit from reviewing (1) injections technique with electromyography or ultrasound guidance, (2) muscles selection and (3) dose of BoNT. In addition, in both scenarios, a holistic approach including drug treatment, retraining and psychological support is valuable in the management of these complex and severe cervical dystonia. PMID:26976927

  19. Structural Insights into the Functional Role of the Hcn Sub-domain of the Receptor-Binding Domain of the Botulinum Neurotoxin Mosaic Serotype C/D

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yanfeng; Gardberg, Anna; Edwards, Tom E.; Sankaran, Banumathi; Robinson, Howard; Varnum, Susan M.; Buchko, Garry W.

    2013-07-01

    Botulinum neurotoxin (BoNT), the causative agent of the deadly neuroparalytic disease botulism, is the most poisonous protein known for humans. Produced by different strains of the anaerobic bacterium Clostridium botulinum, BoNT effects cellular intoxication via a multistep mechanism executed by the three modules of the activated protein. Endocytosis, the first step of cellular intoxication, is triggered by the ~50 kDa, heavy-chain receptor-binding module (HCR) that is specific for a ganglioside and a protein receptor on neuronal cell surfaces. This dual receptor recognition mechanism between BoNT and the host cell’s membrane is well documented and occurs via specific intermolecular interactions with the C-terminal sub-domain, Hcc, of BoNT-HCR. The N-terminal sub-domain of BoNT-HCR, Hcn, comprises ~50% of BoNT-HCR and adopts a B-sheet jelly roll fold. While suspected in assisting cell surface recognition, no unambiguous function for the Hcn sub-domain in BoNT has been indentified. To obtain insights into the potential function of the Hcn sub-domain in BoNT, the first crystal structure of a BoNT with an organic ligand bound to the Hcn sub-domain has been obtained. Here, we describe the crystal structure of BoNT/CD-HCR determined at 1.70 Å resolution with a tetraethylene glycol (PG4) molecule bound in an hydrophobic cleft between B-strands in the B-sheet jelly fold roll of the Hcn sub-domain. The molecule is completely engulfed in the cleft, making numerous hydrophobic (Y932, S959, W966, and D1042) and hydrophilic (S935, W977, L979, N1013, and I1066) contacts with the protein’s side chain and backbone that may mimic in vivo interactions with the phospholipid membranes on neuronal cell surfaces. A sulfate ion was also observed bound to residues T1176, D1177, K1196, and R1243 in the Hcc sub-domain of BoNT/CD-HCR. In the crystal structure of a similar protein, BoNT/D-HCR, a sialic acid

  20. [Botulinum toxin type A in headache treatment : Established and experimental indications].

    Science.gov (United States)

    Gaul, C; Holle-Lee, D; Straube, A

    2016-08-01

    In recent years botulinum toxin type A has been used increasingly more in the treatment of specific headache disorders. Especially regarding chronic migraine with and without combined medication overuse, convincing randomized studies have proven the efficacy of this treatment option and have led to approval for this indication. Regarding other headache entities, such as episodic migraine, tension-type headache, trigeminal autonomic cephalalgia (TAC), neuralgic, neuropathic and myofascial pain, currently available scientific data on the efficacy of botulinum toxin type A are scarce and often ambiguous. The exact underlying mechanisms of the influence of botulinum toxin type A on the pathophysiology of headache are not completely clear but an influence on the release of calcitonin gene-related peptide (CGRP) seems to play a crucial role. This article summarizes the most important studies as well as experiences of treatment with botulinum toxin type A regarding different headache entities. PMID:27300190

  1. Botulinum toxin and its clinical aspects: An overview

    Directory of Open Access Journals (Sweden)

    Shatavisa Mukherjee

    2015-01-01

    Full Text Available Botulinum toxin (BTX, a potent neurotoxin which is produced by the bacterium Clostridium botulinum, consists of eight distinct neurotoxin serotypes referred to as (BTX type-A [BTX-A], B, C, D, E, F, G, H all of which inhibit acetylcholine release at the neuromuscular junction. BTX-A, by blocking acetylcholine release at neuromuscular junctions, accounts for its therapeutic action to relieve dystonia, spasticity, and related disorders. A wide variety of medical conditions such as bruxism, hyperhidrosis, achalasia, focal dystonia, upper motor neuron syndrome, blepharospasm, and chronic migraine are now treated with BTX. The cosmetological applications include correction of lines, creases, and wrinkling all over the face, chin, neck, and chest. Side effects are generally rare and minimal. Injections with BTX-A are well-tolerated. Discovery of further newer indications of this neurotoxin can enlighten the path of research in the field of neuroscience.

  2. Long-lasting attenuation of amygdala-kindled seizures after convection-enhanced delivery of botulinum neurotoxins a and B into the amygdala in rats.

    Science.gov (United States)

    Gasior, Maciej; Tang, Rebecca; Rogawski, Michael A

    2013-09-01

    Botulinum neurotoxins (BoNTs) are well recognized to cause potent, selective, and long-lasting neuroparalytic actions by blocking cholinergic neurotransmission to muscles and glands. There is evidence that BoNT isoforms can also inhibit neurotransmission in the brain. In this study, we examined whether locally delivered BoNT/A and BoNT/B can attenuate kindling measures in amygdala-kindled rats. Male rats were implanted with a combination infusion cannula-stimulating electrode assembly into the right basolateral amygdala. Fully kindled animals received a single infusion of vehicle or BoNT/A or BoNT/B at doses of 1, 3.2, or 10 ng over a 20-minute period by convection-enhanced delivery. Electrographic (EEG) and behavioral kindling measures were determined at selected times during the 3- to 64-day period after the infusion. BoNT/B produced a dose-dependent elevation in after-discharge threshold and duration and a reduction in the seizure stage and duration of behavioral seizures that lasted for up to 50 days after infusion. BoNT/A had similar effects on EEG measures; behavioral seizure measures were also reduced, but the effect did not reach statistical significance. The effects of both toxins on EEG and behavioral measures progressively resolved during the latter half of the observation period. Animals gained weight normally, maintained normal body temperature, and did not show altered behavior. This study demonstrates for the first time that locally delivered BoNTs can produce prolonged inhibition of brain excitability, indicating that they could be useful for the treatment of brain disorders, including epilepsy, that would benefit from long-lasting suppression of neurotransmission within a circumscribed brain region.

  3. Botulinum toxin type A in the treatment of patients with cervical dystonia

    OpenAIRE

    Allison Brashear

    2008-01-01

    Allison BrashearDept of Neurology, Wake Forest University Baptist, Medical Center, Winston Salem, NC, USAAbstract: Dystonia is an involuntary movement involving twisting and turning of agonist and antagonist muscles. Cervical dystonia is isolated to neck musculature. Botulinum toxin type A is a safe and effective treatment of this disabling and often painful syndrome. Three forms of botulinum toxin type A are available worldwide to treat patients with cervical dystonia. This is a review of th...

  4. Induction of an immune response by oral administration of recombinant botulinum toxin.

    OpenAIRE

    Kiyatkin, N; Maksymowych, A B; Simpson, L L

    1997-01-01

    A gene encoding the full-size botulinum neurotoxin serotype C was reconstructed in vector pQE-30 and expressed at high levels in Escherichia coli. Three amino acid mutations (H229-->G, E230-->T, and H233-->N) were generated in the zinc-binding motif, resulting in complete detoxification of the modified recombinant holotoxin. The PCR-amplified wild-type light chain of botulinum neurotoxin serotype C was also expressed in E. coli and used as a control in all experiments. Modified recombinant ho...

  5. Remote Effects of Local Injection of Botulinum Toxin Type A

    Institute of Scientific and Technical Information of China (English)

    Wan Xinhua; Tang Xiaofu; Cui Liying

    2000-01-01

    Objective To assess the seventy and temporal profile of remote effects of botulinum toin type A, BTX-A (Botox from Allergan Inc.,USA and CBTX-A made by Lanzhou Biological Products Institute, China ) injected locally on neuromuscular transmission. Background Recently the local injection of BTX-A has been accepted as a breakthrough in the treatment of a variety of spasmodic disorders, and remote effects of BTX-A on the neuromuscular transmission are concemed. Methods Fourty patients who had enrolled in a prospective open study with Botox or CBTX-A for their movement disorders were studied, 18 cases with Botox and 22 cases with CBTX-A. SFEMG in the extensor digitorum communis muscle or tibialis anterior muscle was performed before and 2-3weeks, 5-8weeks, 4-5 months after injection of Botox or CBTX-A, totally 119 times. Results The significant increase of jitter was demonstrated 2-3weeks after injections in both groups and MCD was in direct proportion to dose of injections.Fiber density value increased at the same time or later and still existed until 4-5 months after injections. Conclusion There are subclinical effects on neuromuscular transmission of remote uninjected muscles after injections of Botox and CBTX A, which indicates that the toxin spread remotely from the site of injection.

  6. Treatment of Raynaud's phenomenon with botulinum toxin type A.

    Science.gov (United States)

    Zhang, Xiaolong; Hu, Yong; Nie, Zhiyu; Song, Ye; Pan, Yougui; Liu, Ying; Jin, Lingjing

    2015-07-01

    Raynaud's phenomenon (RP), an episodic vasospasm of the peripheral arteries, is quite common in general population. The current therapies of RP are limited by efficacy, side effects, and polypharmacy concerns. Botulinum toxin type A (BTX-A) local injections have been reported for the treatment of RP, but the injection sites, concentration and dose of BTX-A were different from each other in previous trials. In addition, so far, there have been no reports concerning local injection of BTX-A in Asian RP patients. Ten patients with RP in China were included in this retrospective study. All the patients had intractable pain and were non-responsive to conservative and/or medical therapy. A patterned BTX-A injection was performed in RP patients, guided by ultrasonography. BTX-A was injected as 20 u/ml devoid of preservatives. Outcomes were measured by ultrasonography, surface temperature, visual analog scale (VAS) for clinical symptoms (pain, numbness, stiffness and swelling), and changes in ulcers or gangrene. Overall, a great improvement in artery flow velocity (P < 0.01), surface temperature (P < 0.01), ulcer and VAS for clinical symptoms, was observed after BTX-A local injection. Complications were very rarely found, and no patients complained of hand weakness and bruise. BTX-A patterned injection guided by ultrasonography might be a useful therapeutic tool in the management of intractable RP.

  7. Potent new small-molecule inhibitor of botulinum neurotoxin serotype A endopeptidase developed by synthesis-based computer-aided molecular design.

    Directory of Open Access Journals (Sweden)

    Yuan-Ping Pang

    Full Text Available Botulinum neurotoxin serotype A (BoNTA causes a life-threatening neuroparalytic disease known as botulism. Current treatment for post exposure of BoNTA uses antibodies that are effective in neutralizing the extracellular toxin to prevent further intoxication but generally cannot rescue already intoxicated neurons. Effective small-molecule inhibitors of BoNTA endopeptidase (BoNTAe are desirable because such inhibitors potentially can neutralize the intracellular BoNTA and offer complementary treatment for botulism. Previously we reported a serotype-selective, small-molecule BoNTAe inhibitor with a K(i (app value of 3.8+/-0.8 microM. This inhibitor was developed by lead identification using virtual screening followed by computer-aided optimization of a lead with an IC(50 value of 100 microM. However, it was difficult to further improve the lead from micromolar to even high nanomolar potency due to the unusually large enzyme-substrate interface of BoNTAe. The enzyme-substrate interface area of 4,840 A(2 for BoNTAe is about four times larger than the typical protein-protein interface area of 750-1,500 A(2. Inhibitors must carry several functional groups to block the unusually large interface of BoNTAe, and syntheses of such inhibitors are therefore time-consuming and expensive. Herein we report the development of a serotype-selective, small-molecule, and competitive inhibitor of BoNTAe with a K(i value of 760+/-170 nM using synthesis-based computer-aided molecular design (SBCAMD. This new approach accounts the practicality and efficiency of inhibitor synthesis in addition to binding affinity and selectivity. We also report a three-dimensional model of BoNTAe in complex with the new inhibitor and the dynamics of the complex predicted by multiple molecular dynamics simulations, and discuss further structural optimization to achieve better in vivo efficacy in neutralizing BoNTA than those of our early micromolar leads. This work provides new insight

  8. Analysis of the neurotoxin complex genes in Clostridium botulinum A1-A4 and B1 strains: BoNT/A3, /Ba4 and /B1 clusters are located within plasmids.

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    Theresa J Smith

    Full Text Available BACKGROUND: Clostridium botulinum and related clostridial species express extremely potent neurotoxins known as botulinum neurotoxins (BoNTs that cause long-lasting, potentially fatal intoxications in humans and other mammals. The amino acid variation within the BoNT is used to categorize the species into seven immunologically distinct BoNT serotypes (A-G which are further divided into subtypes. The BoNTs are located within two generally conserved gene arrangements known as botulinum progenitor complexes which encode toxin-associated proteins involved in toxin stability and expression. METHODOLOGY/PRINCIPAL FINDINGS: Because serotype A and B strains are responsible for the vast majority of human botulism cases worldwide, the location, arrangement and sequences of genes from eight different toxin complexes representing four different BoNT/A subtypes (BoNT/A1-Ba4 and one BoNT/B1 strain were examined. The bivalent Ba4 strain contained both the BoNT/A4 and BoNT/bvB toxin clusters. The arrangements of the BoNT/A3 and BoNT/A4 subtypes differed from the BoNT/A1 strains and were similar to those of BoNT/A2. However, unlike the BoNT/A2 subtype, the toxin complex genes of BoNT/A3 and BoNT/A4 were found within large plasmids and not within the chromosome. In the Ba4 strain, both BoNT toxin clusters (A4 and bivalent B were located within the same 270 kb plasmid, separated by 97 kb. Complete genomic sequencing of the BoNT/B1 strain also revealed that its toxin complex genes were located within a 149 kb plasmid and the BoNT/A3 complex is within a 267 kb plasmid. CONCLUSIONS/SIGNIFICANCE: Despite their size differences and the BoNT genes they contain, the three plasmids containing these toxin cluster genes share significant sequence identity. The presence of partial insertion sequence (IS elements, evidence of recombination/gene duplication events, and the discovery of the BoNT/A3, BoNT/Ba4 and BoNT/B1 toxin complex genes within plasmids illustrate the

  9. High pressure thermal inactivation of Clostridium botulinum type E endospores – kinetic modeling and mechanistic insights

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    Christian Andreas Lenz

    2015-07-01

    Full Text Available Cold-tolerant, neurotoxigenic, endospore forming Clostridium (C. botulinum type E belongs to the non-proteolytic physiological C. botulinum group II, is primarily associated with aquatic environments, and presents a safety risk for seafood. High pressure thermal (HPT processing exploiting the synergistic effect of pressure and temperature can be used to inactivate bacterial endospores.We investigated the inactivation of C. botulinum type E spores by (near isothermal HPT treatments at 300 – 1200 MPa at 30 – 75 °C for 1 s – 10 min. The occurrence of heat and lysozyme susceptible spore fractions after such treatments was determined. The experimental data were modeled to obtain kinetic parameters and represented graphically by isoeffect lines. In contrast to findings for spores of other species and within the range of treatment parameters applied, zones of spore stabilization (lower inactivation than heat treatments alone, large heat susceptible (HPT-induced germinated or lysozyme-dependently germinable (damaged coat layer spore fractions were not detected. Inactivation followed 1st order kinetics. DPA release kinetics allowed for insights into possible inactivation mechanisms suggesting a (poorly effective physiologic-like (similar to nutrient-induced germination at ≤ 450 MPa/≤ 45 °C and non-physiological germination at >500 MPa/>60 – 70 °C.Results of this study support the existence of some commonalities in the HPT inactivation mechanism of C. botulinum type E spores and Bacillus spores although both organisms have significantly different HPT resistance properties. The information presented here contributes to closing the gap in knowledge regarding the HPT inactivation of spore formers relevant to food safety and may help industrial implementation of HPT processing. The markedly lower HPT resistance of C. botulinum type E spores than spores from other C. botulinum types, could allow for the implementation of milder processes without

  10. Presence of antibotulinum neurotoxin antibodies in selected wild canids in Israel.

    Science.gov (United States)

    Steinman, Amir; Millet, Neta; Frenkel, Chana; King, Roni; Shpigel, Nahum Y

    2007-07-01

    Serum samples from 35 golden jackals (Canis aureus syriacus), eight wolves (Canis lupus), and four red foxes (Vulpes vulpes) from various regions of Israel were collected during the years 2001-04 and tested for antibodies to Clostridium botulinum neurotoxin (BoNT) types C and D. Antibodies against BoNT types C and D were detected in 10 (29%) and in 3 (9%) of 35 golden jackals, respectively, using enzyme-linked immunosorbent assay. This report describes detection of anti BoNT antibodies in wild canids other than coyotes (Canis latrans) for the first time and demonstrates that C. botulinum type C is prevalent in Israel. PMID:17699099

  11. Toosendanin interferes with pore formation of botulinum toxin type A in PC12 cell membrane

    Institute of Scientific and Technical Information of China (English)

    Mu-feng LI; Yu-liang SHI

    2006-01-01

    Aim: Botulinum neurotoxins (BoNT) abort the process of neurotransmitter release at presynaptic motor nerve terminals, causing muscle paralysis. The ability of botulinum toxin to produce its effect is dependent on the ability of the light chain to cleave the SNARE proteins associated with transmitter release. Translocation of the light chain protease through the heavy chain-formed channel is a pivotal step in the intoxication process. Toosendanin (TSN), a triterpenoid derivative extracted from a Chinese traditional medicine, has been demonstrated to be an effective cure for experimental botulism. This study was designed to explore the antibotulismic mechanisms of toosendanin. Methods: The inside-out singlechannel recording patch-clamp technique was used to record the BoNT/A-induced currents in the presence and absence of TSN. Results: Channel formation was delayed and the sizes of the channels were reduced in the TSN-treated PC12cell membrane. Conclusion: The antibotulismic effect of TSN might occur via interference with toxin translocation.

  12. Efficacy of Botulinum Toxin Injections in the Treatment of Various Types of Facial Region Disorders

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    Arzu Çoban

    2012-12-01

    Full Text Available OBJECTIVE: Local injection of botulinum toxin is a highly effective treatment option for a wide range of movement disorders and there are reliable sources of information on its indications, effects and safety in clinical practice. In this study, we report our experience with botulinum toxin in the treatment of facial region disorders. METHODS: Patients who had been followed in the Botulinum Toxin Outpatient Clinic of the Neurology Department were retrospectively evaluated. Two preparations of botulinum toxin type A (BT-A were used. The efficacy of BT-A injections was rated according to the improvement in symptoms as follows: marked - 75-100% improvement, good - 50-74%, moderate - 25-49%, and insufficient - less than 25% symptom relief. RESULTS: One hundred eighty-two patients (73 male, 109 female with various facial region disorders were included. The efficacy rates for patients who had very good and good improvement were high in the treatment of blepharospasm, hemifacial spasm, facial synkinesis, and Meige syndrome and moderate for oromandibular dystonia and hypersalivation. Ptosis was the most common side effect. CONCLUSION: According to our results, botulinum toxin was very effective treatment for blepharospasm, Meige syndrome, hemifacial spasm and facial synkinesis, whereas it demonstrated good efficacy in oromandibular dystonia and hypersalivation.

  13. Arrangement of the Clostridium baratii F7 toxin gene cluster with identification of a σ factor that recognizes the botulinum toxin gene cluster promoters.

    Science.gov (United States)

    Dover, Nir; Barash, Jason R; Burke, Julianne N; Hill, Karen K; Detter, John C; Arnon, Stephen S

    2014-01-01

    Botulinum neurotoxin (BoNT) is the most poisonous substances known and its eight toxin types (A to H) are distinguished by the inability of polyclonal antibodies that neutralize one toxin type to neutralize any of the other seven toxin types. Infant botulism, an intestinal toxemia orphan disease, is the most common form of human botulism in the United States. It results from swallowed spores of Clostridium botulinum (or rarely, neurotoxigenic Clostridium butyricum or Clostridium baratii) that germinate and temporarily colonize the lumen of the large intestine, where, as vegetative cells, they produce botulinum toxin. Botulinum neurotoxin is encoded by the bont gene that is part of a toxin gene cluster that includes several accessory genes. We sequenced for the first time the complete botulinum neurotoxin gene cluster of nonproteolytic C. baratii type F7. Like the type E and the nonproteolytic type F6 botulinum toxin gene clusters, the C. baratii type F7 had an orfX toxin gene cluster that lacked the regulatory botR gene which is found in proteolytic C. botulinum strains and codes for an alternative σ factor. In the absence of botR, we identified a putative alternative regulatory gene located upstream of the C. baratii type F7 toxin gene cluster. This putative regulatory gene codes for a predicted σ factor that contains DNA-binding-domain homologues to the DNA-binding domains both of BotR and of other members of the TcdR-related group 5 of the σ70 family that are involved in the regulation of toxin gene expression in clostridia. We showed that this TcdR-related protein in association with RNA polymerase core enzyme specifically binds to the C. baratii type F7 botulinum toxin gene cluster promoters. This TcdR-related protein may therefore be involved in regulating the expression of the genes of the botulinum toxin gene cluster in neurotoxigenic C. baratii.

  14. Characterisation of botulinum toxins type A and B, by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

    NARCIS (Netherlands)

    Baar, B.L.M. van; Hulst, A.G.; Jong, A.L. de; Wils, E.R.J.

    2002-01-01

    A method earlier developed for the mass spectrometric (MS) identification of tetanus toxin (TTx) was applied to botulinum toxins type A and B (BTxA and BTxB). Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. Biologically active BTxA and BTxB

  15. Botulinum Toxin Treatment of Autonomic Disorders: Focal Hyperhidrosis and Sialorrhea.

    Science.gov (United States)

    Hosp, Christine; Naumann, Markus K; Hamm, Henning

    2016-02-01

    Primary focal hyperhidrosis is a common autonomic disorder that significantly impacts quality of life. It is characterized by excessive sweating confined to circumscribed areas, such as the axillae, palms, soles, and face. Less frequent types of focal hyperhidrosis secondary to underlying causes include gustatory sweating in Frey's syndrome and compensatory sweating in Ross' syndrome and after sympathectomy. Approval of onabotulinumtoxinA for severe primary axillary hyperhidrosis in 2004 has revolutionized the treatment of this indication. Meanwhile further type A botulinum neurotoxins like abobotulinumtoxinA and incobotulinumtoxinA, as well as the type B botulinum neurotoxin rimabotulinumtoxinB are successfully used off-label for axillary and various other types of focal hyperhidrosis. For unexplained reasons, the duration of effect differs considerably at different sites. Beside hyperhidrosis, botulinum neurotoxin is also highly valued for the treatment of sialorrhea affecting patients with Parkinson's disease, cerebral palsy, amyotrophic lateral sclerosis, motor neuron disease, and other neurologic conditions. With correct dosing and application, side effects are manageable and transient. PMID:26866492

  16. Botulinum Toxin Treatment of Autonomic Disorders: Focal Hyperhidrosis and Sialorrhea.

    Science.gov (United States)

    Hosp, Christine; Naumann, Markus K; Hamm, Henning

    2016-02-01

    Primary focal hyperhidrosis is a common autonomic disorder that significantly impacts quality of life. It is characterized by excessive sweating confined to circumscribed areas, such as the axillae, palms, soles, and face. Less frequent types of focal hyperhidrosis secondary to underlying causes include gustatory sweating in Frey's syndrome and compensatory sweating in Ross' syndrome and after sympathectomy. Approval of onabotulinumtoxinA for severe primary axillary hyperhidrosis in 2004 has revolutionized the treatment of this indication. Meanwhile further type A botulinum neurotoxins like abobotulinumtoxinA and incobotulinumtoxinA, as well as the type B botulinum neurotoxin rimabotulinumtoxinB are successfully used off-label for axillary and various other types of focal hyperhidrosis. For unexplained reasons, the duration of effect differs considerably at different sites. Beside hyperhidrosis, botulinum neurotoxin is also highly valued for the treatment of sialorrhea affecting patients with Parkinson's disease, cerebral palsy, amyotrophic lateral sclerosis, motor neuron disease, and other neurologic conditions. With correct dosing and application, side effects are manageable and transient.

  17. The use of botulinum toxin type A in cosmetic facial procedures

    NARCIS (Netherlands)

    Jaspers, G. W. C.; Pijpe, J.; Jansma, J.

    2011-01-01

    Over the past decade, facial cosmetic procedures have become more commonplace ill dentistry and oral and maxillofacial surgery. An increasing number of patients seek minimal invasive procedures. One of the most requested procedures is treatment with botulinum toxin type A (BoNTA). Treatment of dynam

  18. Analgesic effects of intra-articular botulinum toxin Type B in a murine model of chronic degenerative knee arthritis pain

    Directory of Open Access Journals (Sweden)

    Stephanie Anderson

    2010-09-01

    Full Text Available Stephanie Anderson1,2, Hollis Krug1,2, Christopher Dorman1, Pari McGarraugh1, Sandra Frizelle1, Maren Mahowald1,21Rheumatology Section, Veteran’s Affairs Medical Center, Minneapolis, Minnesota; 2Division of Rheumatology and Autoimmune Diseases, University of Minnesota Medical School, Minneapolis, Minnesota, USAObjective: To evaluate the analgesic effectiveness of intra-articular botulinum toxin Type B (BoNT/B in a murine model of chronic degenerative arthritis pain.Methods and materials: Chronic arthritis was produced in adult C57Bl6 mice by intra-articular injection of Type IV collagenase into the left knee. Following induction of arthritis, the treatment group received intra-articular BoNT/B. Arthritic control groups were treated with intra-articular normal saline or sham injections. Pain behavior testing was performed prior to arthritis, after induction of arthritis, and following treatments. Pain behavior measures included analysis of gait impairment (spontaneous pain behavior and joint tenderness evaluation (evoked pain response. Strength was measured as ability to grasp and cling.Results: Visual gait analysis showed significant impairment of gait in arthritic mice that improved 43% after intra-articular BoNT/B, demonstrating a substantial articular analgesic effect. Joint tenderness, measured with evoked pain response scores, increased with arthritis induction and decreased 49.5% after intra-articular BoNT/B treatment. No improvement in visual gait scores or decrease in evoked pain response scores were found in the control groups receiving intra-articular normal saline or sham injections. Intra-articular BoNT/B was safe, and no systemic effects or limb weakness was noted.Conclusions: This study is the first report of intra-articular BoNT/B for analgesia in a murine model of arthritis pain. The results of this study validate prior work using intra-articular neurotoxins in murine models. Our findings show chronic degenerative arthritis

  19. Biodiversity of Clostridium botulinum Type E Strains Isolated from Fish and Fishery Products

    OpenAIRE

    HyytiÀ, E.; Hielm, S.; Björkroth, J.; Korkeala, H.

    1999-01-01

    The genetic biodiversity of Clostridium botulinum type E strains was studied by pulsed-field gel electrophoresis (PFGE) with two macrorestriction enzymes (SmaI-XmaI and XhoI) and by randomly amplified polymorphic DNA (RAPD) analysis with two primers (OPJ 6 and OPJ 13) to characterize 67 Finnish isolates from fresh fish and fishery products, 15 German isolates from farmed fish, and 10 isolates of North American or North Atlantic origin derived mainly from different types of seafood. The effect...

  20. The Antigenicity Analysis of Botulinum Toxin of Clostridium botulinum Type C%C型肉毒梭菌肉毒毒素的抗原性分析

    Institute of Scientific and Technical Information of China (English)

    王娟娟; 刘书东; 李剑; 刘志龙; 陈根元; 曹玉华; 李莲瑞

    2012-01-01

    [目的]通过对分离纯化的C型肉毒毒素抗原性分析,提供快速鉴定动物因C型肉毒梭菌中毒的理论基础.[方法]对分离纯化的C型肉毒梭菌的肉毒毒素灭活后通过SDS - PAGE测定浓度,确定浓度后与弗氏佐剂乳化后免疫家兔,免疫结束后采家兔血液并分离血清.[结果]将不同稀释倍数的肉毒毒素经SDS -PAGE电泳分离后,转移至HybondNC膜,家兔三免后提取的血清作为一抗,用二抗进行Western blotting检测,大约在98.0和53.0 kDa处出现特异反应带;抗原与抗体在琼脂凝胶上结合时,会形成清晰的沉淀带.[结论]分离纯化的C型肉毒毒素,经免疫后家兔可见:C型肉毒毒素具有免疫原性和反应原性,抗体的效价为1∶8.%[Objective ] The study aims to provide the theoretical basis for quick identification of Clostridium botulinum type C through the separation and purification of type C botulinum toxin antigenic analysis. [ Method ] The experiments were carried out by separating and purifyinng Clostridium botulinum type C in the blood collected from Freunds adjuvant emulsified rabbit and the botulinum toxin was inactivated to determine the concentration by SDS - PAGE. The rabbit blood was collected after immunization and the serum was separated. [ Result ] Different dilutions of botulinum toxin were separated by SDS - PAGE electrophoresis, transferred to HybondNC film, and the three free extracted serum from rabbits was used as primary antibody. The secondary antibody was used for Western blotting analysis. At approximately 98.0 and 53.0 kDa, specific reaction zone appeared; antigens and antibodies in agar gel when combined will form a clear precipitation band. [Conclusion]The immunized rabbits, whose botulinum toxin type C was separated and purified by us, are still there. C botulinum toxin has the characteristics of immunogenicity and reactogenicity, and its antibody ti-ter is 1=8.

  1. Botulinum Toxin Type A as a Therapeutic Agent against Headache and Related Disorders

    Directory of Open Access Journals (Sweden)

    Siro Luvisetto

    2015-09-01

    Full Text Available Botulinum neurotoxin A (BoNT/A is a toxin produced by the naturally-occurring Clostridium botulinum that causes botulism. The potential of BoNT/A as a useful medical intervention was discovered by scientists developing a vaccine to protect against botulism. They found that, when injected into a muscle, BoNT/A causes a flaccid paralysis. Following this discovery, BoNT/A has been used for many years in the treatment of conditions of pathological muscle hyperactivity, like dystonias and spasticities. In parallel, the toxin has become a “glamour” drug due to its power to ward off facial wrinkles, particularly frontal, due to the activity of the mimic muscles. After the discovery that the drug also appeared to have a preventive effect on headache, scientists spent many efforts to study the potentially-therapeutic action of BoNT/A against pain. BoNT/A is effective at reducing pain in a number of disease states, including cervical dystonia, neuropathic pain, lower back pain, spasticity, myofascial pain and bladder pain. In 2010, regulatory approval for the treatment of chronic migraine with BoNT/A was given, notwithstanding the fact that the mechanism of action is still not completely elucidated. In the present review, we summarize experimental evidence that may help to clarify the mechanisms of action of BoNT/A in relation to the alleviation of headache pain, with particular emphasis on preclinical studies, both in animals and humans. Moreover, we summarize the latest clinical trials that show evidence on headache conditions that may obtain benefits from therapy with BoNT/A.

  2. Botulinum Toxin Type a as a Therapeutic Agent against Headache and Related Disorders.

    Science.gov (United States)

    Luvisetto, Siro; Gazerani, Parisa; Cianchetti, Carlo; Pavone, Flaminia

    2015-09-01

    Botulinum neurotoxin A (BoNT/A) is a toxin produced by the naturally-occurring Clostridium botulinum that causes botulism. The potential of BoNT/A as a useful medical intervention was discovered by scientists developing a vaccine to protect against botulism. They found that, when injected into a muscle, BoNT/A causes a flaccid paralysis. Following this discovery, BoNT/A has been used for many years in the treatment of conditions of pathological muscle hyperactivity, like dystonias and spasticities. In parallel, the toxin has become a "glamour" drug due to its power to ward off facial wrinkles, particularly frontal, due to the activity of the mimic muscles. After the discovery that the drug also appeared to have a preventive effect on headache, scientists spent many efforts to study the potentially-therapeutic action of BoNT/A against pain. BoNT/A is effective at reducing pain in a number of disease states, including cervical dystonia, neuropathic pain, lower back pain, spasticity, myofascial pain and bladder pain. In 2010, regulatory approval for the treatment of chronic migraine with BoNT/A was given, notwithstanding the fact that the mechanism of action is still not completely elucidated. In the present review, we summarize experimental evidence that may help to clarify the mechanisms of action of BoNT/A in relation to the alleviation of headache pain, with particular emphasis on preclinical studies, both in animals and humans. Moreover, we summarize the latest clinical trials that show evidence on headache conditions that may obtain benefits from therapy with BoNT/A.

  3. Botulinum Toxin Type a as a Therapeutic Agent against Headache and Related Disorders.

    Science.gov (United States)

    Luvisetto, Siro; Gazerani, Parisa; Cianchetti, Carlo; Pavone, Flaminia

    2015-09-01

    Botulinum neurotoxin A (BoNT/A) is a toxin produced by the naturally-occurring Clostridium botulinum that causes botulism. The potential of BoNT/A as a useful medical intervention was discovered by scientists developing a vaccine to protect against botulism. They found that, when injected into a muscle, BoNT/A causes a flaccid paralysis. Following this discovery, BoNT/A has been used for many years in the treatment of conditions of pathological muscle hyperactivity, like dystonias and spasticities. In parallel, the toxin has become a "glamour" drug due to its power to ward off facial wrinkles, particularly frontal, due to the activity of the mimic muscles. After the discovery that the drug also appeared to have a preventive effect on headache, scientists spent many efforts to study the potentially-therapeutic action of BoNT/A against pain. BoNT/A is effective at reducing pain in a number of disease states, including cervical dystonia, neuropathic pain, lower back pain, spasticity, myofascial pain and bladder pain. In 2010, regulatory approval for the treatment of chronic migraine with BoNT/A was given, notwithstanding the fact that the mechanism of action is still not completely elucidated. In the present review, we summarize experimental evidence that may help to clarify the mechanisms of action of BoNT/A in relation to the alleviation of headache pain, with particular emphasis on preclinical studies, both in animals and humans. Moreover, we summarize the latest clinical trials that show evidence on headache conditions that may obtain benefits from therapy with BoNT/A. PMID:26404377

  4. Application of Botulinum Toxin in Pain Management

    OpenAIRE

    Sim, Woo Seog

    2011-01-01

    Botulinum toxin has been used for the treatment of many clinical disorders by producing temporary skeletal muscle relaxation. In pain management, botulinum toxin has demonstrated an analgesic effect by reducing muscular hyperactivity, but recent studies suggest this neurotoxin could have direct analgesic mechanisms different from its neuromuscular actions. At the moment, botulinum toxin is widely investigated and used in many painful diseases such as myofascial syndrome, headaches, arthritis,...

  5. Flagellin Diversity in Clostridium botulinum Groups I and II: a New Strategy for Strain Identification▿

    Science.gov (United States)

    Paul, Catherine J.; Twine, Susan M.; Tam, Kevin J.; Mullen, James A.; Kelly, John F.; Austin, John W.; Logan, Susan M.

    2007-01-01

    Strains of Clostridium botulinum are traditionally identified by botulinum neurotoxin type; however, identification of an additional target for typing would improve differentiation. Isolation of flagellar filaments and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that C. botulinum produced multiple flagellin proteins. Nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) analysis of in-gel tryptic digests identified peptides in all flagellin bands that matched two homologous tandem flagellin genes identified in the C. botulinum Hall A genome. Designated flaA1 and flaA2, these open reading frames encode the major structural flagellins of C. botulinum. Colony PCR and sequencing of flaA1/A2 variable regions classified 80 environmental and clinical strains into group I or group II and clustered isolates into 12 flagellar types. Flagellar type was distinct from neurotoxin type, and epidemiologically related isolates clustered together. Sequencing a larger PCR product, obtained during amplification of flaA1/A2 from type E strain Bennett identified a second flagellin gene, flaB. LC-MS analysis confirmed that flaB encoded a large type E-specific flagellin protein, and the predicted molecular mass for FlaB matched that observed by SDS-PAGE. In contrast, the molecular mass of FlaA was 2 to 12 kDa larger than the mass predicted by the flaA1/A2 sequence of a given strain, suggesting that FlaA is posttranslationally modified. While identification of FlaB, and the observation by SDS-PAGE of different masses of the FlaA proteins, showed the flagellin proteins of C. botulinum to be diverse, the presence of the flaA1/A2 gene in all strains examined facilitates single locus sequence typing of C. botulinum using the flagellin variable region. PMID:17351097

  6. Botulinum toxin has an increased effect when targeted toward the muscle's endplate zone: a high-density surface EMG guided study

    NARCIS (Netherlands)

    Lapatki, B.G.; Dijk, J.P. van; Warrenburg, B.P.C. van de; Zwarts, M.J.

    2011-01-01

    OBJECTIVES: To compare the effect of endplate-targeted injections of a low Botulinum neurotoxin type A (BoNT-A) dose with that of injections at defined distances from the motor endplate zone. METHODS: In eight healthy volunteers, the main endplate zones of the right and left extensor digitorum brevi

  7. Jaw-opening oromandibular dystonia secondary to Wilson's Disease treated with botulinum toxin type A

    Directory of Open Access Journals (Sweden)

    Hélio A.G. Teive

    2012-06-01

    Full Text Available We have reported a case series of five patients with jaw-opening oromandibular dystonia secondary to Wilson's disease (WD, in which the patients were treated with botulinum toxin type A (BTX-A. In all cases, dystonia score was partially reduced three weeks after injections. The most common side effect was transient mild dysphagia. This preliminary study showed that jaw-opening oromandibular dystonia in WD may be partially responsive to the use of BTX-A.

  8. Assessment of intralesional injection of botulinum toxin type A injection for hypertrophic scars

    Directory of Open Access Journals (Sweden)

    Alhasan M Elhefnawy

    2016-01-01

    Full Text Available Background: Hypertrophic scars are dermal fibroproliferative disorders that typically develop after a skin injury heals. They can cause physical, psychological, and cosmetic problems. The management of such scars remains a matter of debate due to lack of effective treatment methods and the inability to prevent recurrences. Recent reports have demonstrated that botulinum toxin type A improves wound healing so it may play a role in treating hypertrophic scars. Aims: We assessed the effectiveness of intralesional botulinum toxin type A injection for treating hypertrophic scars. Methods: This prospective clinical study included twenty patients with hypertrophic scars. Intralesional injection of botulinum toxin type A was given once a month for three months with a follow-up period of six months. Each lesion was injected until slight blanching occurred. Therapeutic satisfaction of the patient and physician were recorded. Lesions were assessed for erythema, itching and pliability. Each item was assessed on a 5-point scale. Results: Therapeutic satisfaction was recorded as 'good' in 14 patients and 'excellent' in the remaining six. The mean erythema score decreased from 3.2 to 1.0, the mean pliability score from 3.3 to 0.8 and the mean itching score from 2.7 to 0.7. All of these were statistically significant. Limitations: A larger sample size and longer follow-up period would have given a better evaluation but was not feasible due to the high expenses involved. Conclusion: Botulinum toxin type A is a novel and promising therapy for hypertrophic scars with few side effects.

  9. Clinical and image improvement of Raynaud's phenomenon after botulinum toxin type A treatment.

    Science.gov (United States)

    Zhao, HongMei; Lian, YaJun

    2015-08-01

    Raynaud's phenomenon is often accompanied by pain, digital ulceration and compromised daily activities. Pharmacological therapy or sympathectomies have been administered to diminish these symptoms but existing treatments are not invariably efficacious. A recent case series has described the use of botulinum toxin type A in the treatment of Raynaud's phenomenon. We report two patients with severe or mild Raynaud's phenomenon who were injected with BTX-A; both of whom experienced clinical and image improvement after treatment.

  10. Type A botulinum toxin: a new treatment for axillary and palmar hyperhidrosis.

    Science.gov (United States)

    Rusciani, Luigi; Severino, Enzo; Rusciani, Antonio

    2002-09-01

    Hyperhidrosis is an invalidating condition, and one that is difficult to treat. It is characterized by an excessive and uncontrolled production of sweat by the sweat glands, often causing psychological, social, and occupational problems for the patient. Hyperhidrosis can be distinguished in two forms: idiopathic (of unknown etiology), or secondary, due to an alteration of the endocrine system (ex: hyperthyroidism, neuropathy, neoplasia etc.) It is found in about 0.3-0.5% of the population and can be localized (axillary, palmar, plantar, facial) or diffused. The subcutaneous injection of type A botulinum toxin, until now used only for the treatment of blepharospasm or hemifacial spasm, has shown to be a useful treatment for localized hyperhidrosis. The objective of the authors is to evaluate the therapeutic efficacy, safety, and management of botulinum toxin treatment in patients affected with axillary or palmar hyperhidrosis resistant to conventional therapies. PMID:12847738

  11. Biodiversity of Clostridium botulinum Type E Associated with a Large Outbreak of Botulism in Wildlife from Lake Erie and Lake Ontario ▿

    OpenAIRE

    Hannett, George E.; Stone, Ward B.; Davis, Stephen W.; Wroblewski, Danielle

    2010-01-01

    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...

  12. Biodiversity of Clostridium botulinum type E associated with a large outbreak of botulism in wildlife from Lake Erie and Lake Ontario.

    Science.gov (United States)

    Hannett, George E; Stone, Ward B; Davis, Stephen W; Wroblewski, Danielle

    2011-02-01

    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 detected in 110 samples from birds, 12 samples from fish, and 2 samples from mammals. Sediment samples from Lake Erie were also examined for the presence of C. botulinum. Fifteen of 17 sediment samples were positive for the presence of C. botulinum type E. Eighty-one C. botulinum isolates were obtained from plants, animals, and sediments; of these isolates, 44 C. botulinum isolates produced type E toxin, as determined by mouse bioassay, while the remaining 37 isolates were not toxic for mice. All toxin-producing isolates were typed by RAPD; that analysis showed 12 different RAPD types and multiple subtypes. Our study thus demonstrates that multiple genetically distinct strains of C. botulinum were involved in the present outbreak of wildlife botulism. We found that C. botulinum type E is present in the sediments of Lake Erie and that a large range of bird and fish species is affected. PMID:21115703

  13. Application of botulinum toxin in pain management.

    Science.gov (United States)

    Sim, Woo Seog

    2011-03-01

    Botulinum toxin has been used for the treatment of many clinical disorders by producing temporary skeletal muscle relaxation. In pain management, botulinum toxin has demonstrated an analgesic effect by reducing muscular hyperactivity, but recent studies suggest this neurotoxin could have direct analgesic mechanisms different from its neuromuscular actions. At the moment, botulinum toxin is widely investigated and used in many painful diseases such as myofascial syndrome, headaches, arthritis, and neuropathic pain. Further studies are needed to understand the exact analgesic mechanisms, efficacy and complications of botulinum toxin in chronic pain disorders.

  14. Use of Clostridium botulinum toxin in gastrointestinalmotility disorders in children

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    More than a century has elapsed since the identificationof Clostridia neurotoxins as the cause of paralyticdiseases. Clostridium botulinum is a heterogeneousgroup of Gram-positive, rod-shaped, spore-forming,obligate anaerobic bacteria that produce a potentneurotoxin. Eight different Clostridium botulinumneurotoxins have been described (A-H) and 5 of thosecause disease in humans. These toxins cause paralysisby blocking the presynaptic release of acetylcholine atthe neuromuscular junction. Advantage can be taken ofthis blockade to alleviate muscle spams due to excessiveneural activity of central origin or to weaken a musclefor treatment purposes. In therapeutic applications,minute quantities of botulinum neurotoxin type A areinjected directly into selected muscles. The Food andDrug Administration first approved botulinum toxin (BT)type A in 1989 for the treatment of strabismus andblepharospasm associated with dystonia in patients 12years of age or older. Ever since, therapeutic applicationsof BT have expanded to other systems, including thegastrointestinal tract. Although only a single fatalityhas been reported to our knowledge with use of BTfor gastroenterological conditions, there are significantcomplications ranging from minor pain, rash and allergicreactions to pneumothorax, bowel perforation andsignificant paralysis of tissues surrounding the injection(including vocal cord paralysis and dysphagia). Thiseditorial describes the clinical experience and evidencefor the use BT in gastrointestinal motility disorders inchildren.

  15. Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

    NARCIS (Netherlands)

    Baar, B.L.M. van; Hulst, A.G.; Jong, A.L. de; Wils, E.R.J.

    2004-01-01

    In a follow-up of the earlier characterisation of botulinum toxins type A and B (BTxA and BTxB) by mass spectrometry (MS), types C, D, E, and F (BTxC, BTxD, BTxE, BTxF) were now investigated. Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. B

  16. Botulinum toxin type A chemodenervation treatment in spastic forms of cerebral palsy

    OpenAIRE

    A. L. Kurenkov; S. S. Nikitin; A. R. Artemenko; B. I. Bursagova; Kuzenkova, L.M.; Petrova, S. A.; O. A. Klochkova; A. M. Mamedyarov

    2015-01-01

    Cerebral palsy (CP) is one of the most serious outcomes of the perinatal lesion of central nervous system and the most common reason for neurological disability in children. Being the key cause of pathological dynamic stereotypes that frequently result in pathological posture and contractures, spasticity is critically important for CP. The use of botulinum toxin type A (BTA) in complex treatment 2-6 years old CP patients allows significantly to improve motor abilities, help to change the surg...

  17. Role of Botulinum Toxin Type-A (BTX-A) in the Management of Trigeminal Neuralgia

    OpenAIRE

    Gaurav Verma

    2013-01-01

    Trigeminal neuralgia (TN) is a clinical condition characterized by paroxysmal attacks of severe and electric shock-like pain along the distribution of one or more branches of the trigeminal nerve. Various medicinal or surgical modalities have been employed in the past with variable success. Newer methods were tried in search of permanent cure or long-lasting pain relief. The purpose of this paper is to present the review of the literature regarding the use of botulinum toxin type-A (BTX-A) in...

  18. Quantification of Clostridium botulinum Toxin Gene Expression by Competitive Reverse Transcription-PCR

    OpenAIRE

    McGrath, S.; Dooley, J. S. G.; Haylock, R. W.

    2000-01-01

    Clostridium botulinum produces a characteristic botulinum neurotoxin which can cause an often fatal neuroparalytic condition known as botulism. Although food-borne botulism is rare, critical screening by food companies is necessary to ensure that food products are safe. At present, the food industry assesses the risks of botulinum neurotoxin production by challenge testing to check any new food products and to check the efficacy of new storage regimes. Challenge testing involves artificial in...

  19. Neurophysiological changes induced by the botulinum toxin type A injection in children with cerebral palsy.

    Science.gov (United States)

    Frascarelli, Flaminia; Di Rosa, Giuseppe; Bisozzi, Eleonora; Castelli, Enrico; Santilli, Valter

    2011-01-01

    In the last few years botulinum toxin type A (BTX-A) has been widely used in the management of spasticity in children with cerebral palsy in order to reduce hypertonicity and improve functional outcomes enhancing motor skill development. The botulinum toxin injection seems to interact with intrafusal and extrafusal fibers producing a reduction of hypertone both through synaptic blockade and inhibition of stretch reflex loop and these changes may influence not only the spinal cord but also the central nervous system (CNS). The purpose of our study was to determine the neurophysiological changes induced by the BTX-A through an evaluation of cortical somatosensory Evoked Potential (SEP) and Soleus H wave, that is the index of excitability of stretch reflex loop. Eighteen children with Cerebral Palsy (CP), aged between 5 and 12, were recruited at Children's Hospital "Bambino Gesù" of Rome. All children were evaluated with appropriate clinical scales before and 1 month after the BTX-A injection. Neurophysiological measurements were performed before, and 1 month after botulinum toxin injection through lower limb SEPs, M-wave and Soleus H wave recording. After the injection the results showed a statistically significant improvement both of clinical scales and the neurophysiological variables. These findings suggest that spasticity itself can be considered as a factor affecting the cortical SEPs. And even though it seems that BTX-A does not have any direct central effect on sensory pathways we suppose an indirect mechanism on modulation of afferent fibers Ia due to the modification induced by BTX-A to central loop reflex.

  20. Quantum dot immunoassays in renewable surface column and 96-well plate formats for the fluorescence detection of Botulinum neurotoxin using high-affinity antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Warner, Marvin G.; Grate, Jay W.; Tyler, Abby J.; Ozanich, Richard M.; Miller, Keith D.; Lou, Jianlong; Marks, James D.; Bruckner-Lea, Cindy J.

    2009-09-01

    A fluorescence sandwich immunoassay using high affinity antibodies and quantum dot (QD) reporters has been developed for detection of botulinum toxin serotype A (BoNT/A). For the development of the assay, a nontoxic recombinant fragment of the holotoxin (BoNT/A-HC-fragment) has been used as a structurally valid simulant for the full toxin molecule. The antibodies used, AR4 and RAZ1, bind to nonoverlapping epitopes present on both the full toxin and on the recombinant fragment. In one format, the immunoassay is carried out in a 96-well plate with detection in a standard plate reader. Detection down to 31 pM of the BoNT/Hc-fragment was demonstrated with a total incubation time of 3 hours, using AR4 as the capture antibody and QD-coupled RAZ1 as the reporter. In a second format, the AR4 capture antibody was coupled to Sepharose beads, and the immunochemical reactions were carried out in microcentrifuge tubes with an incubation time of 1 hour. These beads were subsequently captured and concentrated in a rotating rod “renewable surface” flow cell as part of a sequential injection fluidic system. This flow cell was equipped with a fiber optic system for fluorescence measurements. In PBS buffer solution matrix, the BoNT/A-HC-fragment was detected to concentrations as low as 5 pM using the fluidic measurement approach.

  1. Anatomic considerations in botulinum toxin type A therapy for spasmodic dysphonia.

    Science.gov (United States)

    Castellanos, P F; Gates, G A; Esselman, G; Song, F; Vannier, M W; Kuo, M

    1994-06-01

    Chemodenervation by injection of botulinum toxin type A into the vocal fold(s) has become the preferred treatment for patients with adductor spasmodic dysphonia. Injection may be done either perorally or transcutaneously; each method has its advocates and advantages. The authors have used the transcutaneous transcricothyroid membrane route exclusively with satisfactory results in more than 50 patients. Temporary breathliness and aspiration are common. The preferred injection site should be as close as possible to the motor end plates of the affected muscle. The thyroarytenoid muscle end plates are distributed throughout the muscle, whereas in the lateral cricoarytenoid muscle they are located in band in the center of the muscle. The transcutaneous injection site is below and posterior to the midpoint of the vibrating vocal fold as visualized by indirect laryngoscopy. The proximity of this site to the lateral cricoarytenoid muscle suggests that postinjection breathiness and aspiration may be related to spread of botulinum toxin type A to the lateral cricoarytenoid muscle. However, it is likely that thyroarytenoid muscle paresis is mainly responsible for this side effect and that the rapid clearing of the breathy dysphonia in the face of prolonged relief of spasmodic dysphonia symptoms suggests the action of an adaptive neural response, such as axonal sprouting. Further research of this subject is warranted. PMID:8196438

  2. Development of an Innovative in Vitro Potency Assay for Anti-Botulinum Antitoxins.

    Science.gov (United States)

    Rosen, Osnat; Ozeri, Eyal; Barnea, Ada; David, Alon Ben; Zichel, Ran

    2016-09-24

    Botulinum neurotoxins are bacterial proteins that cause botulism, a life-threatening disease. Therapy relies mostly on post-intoxication antibody treatment. The only accepted method to measure the potency of, and to approve, antitoxin preparations is the mouse lethality neutralization bioassay. However, this assay is time-consuming, labor-intensive, costly, and raises ethical issues related to the large numbers of laboratory animals needed. Until now, all efforts to develop an alternative in vitro assay have not provided a valid replacement to the mouse potency assay. In the present study, we report the development of an innovative in vitro assay for determining botulinum antitoxin potency, using botulinum type B as a model. The concept of the assay is to mimic two fundamental steps in botulinum intoxication: receptor binding and catalytic activity. By simulating these steps in vitro we were able to accurately determine the potency of antitoxin preparations. The reproducibility of the assay was high with a CV vitro assay highly correlated with that measured by the standard in vivo mouse assay (r = 0.9842, p vitro assay has the potential to be considered, after validation, as a replacement to the mouse assay for quantitating neutralizing antibody concentrations in pharmaceutical botulinum antitoxin preparations. Future adoption of this in vitro assay would minimize the use of laboratory animals, speed up the time, and reduce the cost of botulinum antitoxin approval.

  3. Ineffectiveness of 3,4-diaminopyridine as a therapy for type C botulism.

    Science.gov (United States)

    Siegel, L.S.; Price, J.I.

    1987-01-01

    Clostridium botulinum neurotoxins inhibit acetylcholine release at neuromuscular junctions. Agents stimulating neurotransmitter efflux, such as 3,4-diaminopyridine (3,4-DAP), could be useful for botulism therapy. Treatment with 3,4-DAP (8 mg/kg hourly, beginning 3 hr after toxin injection) failed to increase the survival times of mice receiving 10, 20 or 40 LD50 type C, but did prolong the survival of those receiving 20 LD50 type A. This difference in 3,4-DAP efficacy may reflect variations in the molecular mechanism of action of types A and C botulinum neurotoxins.

  4. Evaluating the synergistic neutralizing effect of anti-botulinum oligoclonal antibody preparations.

    Directory of Open Access Journals (Sweden)

    Eran Diamant

    Full Text Available Botulinum neurotoxins (BoNT are considered some of the most lethal known substances. There are seven botulinum serotypes, of which types A, B and E cause most human botulism cases. Anti-botulinum polyclonal antibodies (PAbs are currently used for both detection and treatment of the disease. However, significant improvements in immunoassay specificity and treatment safety may be made using monoclonal antibodies (MAbs. In this study, we present an approach for the simultaneous generation of highly specific and neutralizing MAbs against botulinum serotypes A, B, and E in a single process. The approach relies on immunization of mice with a trivalent mixture of recombinant C-terminal fragment (Hc of each of the three neurotoxins, followed by a parallel differential robotic hybridoma screening. This strategy enabled the cloning of seven to nine MAbs against each serotype. The majority of the MAbs possessed higher anti-botulinum ELISA titers than anti-botulinum PAbs and had up to five orders of magnitude greater specificity. When tested for their potency in mice, neutralizing MAbs were obtained for all three serotypes and protected against toxin doses of 10 MsLD50-500 MsLD50. A strong synergistic effect of up to 400-fold enhancement in the neutralizing activity was observed when serotype-specific MAbs were combined. Furthermore, the highly protective oligoclonal combinations were as potent as a horse-derived PAb pharmaceutical preparation. Interestingly, MAbs that failed to demonstrate individual neutralizing activity were observed to make a significant contribution to the synergistic effect in the oligoclonal preparation. Together, the trivalent immunization strategy and differential screening approach enabled us to generate highly specific MAbs against each of the A, B, and E BoNTs. These new MAbs may possess diagnostic and therapeutic potential.

  5. Effects of botulinum toxin type A on healing of injured skeletal muscles

    Directory of Open Access Journals (Sweden)

    Shokravi Ramin

    2007-01-01

    Full Text Available Objectives: (1 Evaluation of microscopic healing of skeletal muscle fibers after injuries, especially the arrangement of new muscle fibers and scar tissue diameter in the injury region. (2 Evaluation of alterations in microscopy of the healing procedure within skeletal muscles after injury following botulinum toxin type A (BTX -A induced muscle immobilization. Materials and Methods: The study was done on 12 white lab rabbits of either sex in a 6-month period. Results: The immobilization of skeletal muscle fibers as a result of the use of BTX-A after injury caused a qualitative increase in fibrous tissue formation in the area of injury, and the BTX-A-induced immobilization for a period of 6 months led to muscle atrophy.

  6. Botulinum toxin type A in the treatment of painful adductor muscle contracture after total hip arthroplasty.

    Science.gov (United States)

    Santamato, Andrea; Ranieri, Maurizio; Panza, Francesco; Solfrizzi, Vincenzo; Frisardi, Vincenza; Lapenna, Luisa Maria; Moretti, Biagio; Fiore, Pietro

    2009-10-01

    Painful adductor muscle contracture is an important cause of failure during rehabilitation following total hip arthroplasty (THA). Adductor muscle contracture may be caused by postoperative muscle retractions, adhesive capsulitis, postoperative leg-length inequalities caused by implant failure, or preexisting hip pathologies. A 34-year-old woman experienced a persistent painful contracture into the left adductor magnus muscle after THA. She had no leg-length inequalities and, according to the Medical Research Council scale (grades 0-5), muscle strength of the quadriceps was 5/5 for the right side and 3/5 for the left. The degree of functionality according to the Harris hip score (HHS) was 16/100 in the left hip. The pain level, measured with the visual analog scale (VAS), was 7/10. The patient was unable to fully adhere to the rehabilitation program and walked with a limp during the stance phase of gait. After 7 days of treatment with injections of botulinum toxin type A into the left adductor magnus muscle (dose, 150 UM) and subsequent rehabilitation, a great reduction of painful contracture was observed (VAS score, 2/10). The procedure was well tolerated and no adverse effects were noted. After 20 days, hip articular range of motion and gait had improved (HHS score, 75/100). The clinical effects of botulinum toxin type A were present at 2-month follow-up. This treatment may be a viable alternative for the management of painful adductor muscle contracture after THA, without significant side effects. PMID:19824593

  7. Botulinum Toxin in Pediatric Neurology

    OpenAIRE

    Moawad, Eman M. I.; Abdallah, Enas Abdallah Ali

    2015-01-01

    Botulinum neurotoxins are natural molecules produced by anaerobic spore-forming bacteria called Clostradium boltulinum. The toxin has a peculiar mechanism of action by preventing the release of acetylcholine from the presynaptic membrane. Consequently, it has been used in the treatment of various neurological conditions related to muscle hyperactivity and/or spasticity. Also, it has an impact on the autonomic nervous system by acting on smooth muscle, leading to its use in the management of p...

  8. Botulinum toxin type A products are not interchangeable: a review of the evidence

    Directory of Open Access Journals (Sweden)

    Brin MF

    2014-10-01

    Full Text Available Mitchell F Brin,1,2 Charmaine James,3 John Maltman1 1Allergan, Inc., Irvine, CA, USA; 2Department of Neurology, University of California, Irvine, CA, USA; 3Allergan, Marlow, UKAbstract: Botulinum toxin type A (BoNTA products are injectable biologic medications derived from Clostridium botulinum bacteria. Several different BoNTA products are marketed in various countries, and they are not interchangeable. Differences between products include manufacturing processes, formulations, and the assay methods used to determine units of biological activity. These differences result in a specific set of interactions between each BoNTA product and the tissue injected. Consequently, the products show differences in their in vivo profiles, including preclinical dose response curves and clinical dosing, efficacy, duration, and safety/adverse events. Most, but not all, published studies document these differences, suggesting that individual BoNTA products act differently depending on experimental and clinical conditions, and these differences may not always be predictable. Differentiation through regulatory approvals provides a measure of confidence in safety and efficacy at the specified doses for each approved indication. Moreover, the products differ in the amount of study to which they have been subjected, as evidenced by the number of publications in the peer-reviewed literature and the quantity and quality of clinical studies. Given that BoNTAs are potent biological products that meet important clinical needs, it is critical to recognize that their dosing and product performance are not interchangeable and each product should be used according to manufacturer guidelines.Keywords: onabotulinumtoxinA, abobotulinumtoxinA, incobotulinumtoxinA, glabellar lines, non-interchangeability

  9. Biodiversity of Clostridium botulinum type E strains isolated from fish and fishery products.

    Science.gov (United States)

    Hyytiä, E; Hielm, S; Björkroth, J; Korkeala, H

    1999-05-01

    The genetic biodiversity of Clostridium botulinum type E strains was studied by pulsed-field gel electrophoresis (PFGE) with two macrorestriction enzymes (SmaI-XmaI and XhoI) and by randomly amplified polymorphic DNA (RAPD) analysis with two primers (OPJ 6 and OPJ 13) to characterize 67 Finnish isolates from fresh fish and fishery products, 15 German isolates from farmed fish, and 10 isolates of North American or North Atlantic origin derived mainly from different types of seafood. The effects of fish species, processing, and geographical origin on the epidemiology of the isolates were evaluated. Cluster analysis based on macrorestriction profiles was performed to study the genetic relationships of the isolates. PFGE and RAPD analyses were combined and resulted in the identification of 62 different subtypes among the 92 type E isolates analyzed. High genetic biodiversity among the isolates was observed regardless of their source. Finnish and North American or North Atlantic isolates did not form distinctly discernible clusters, in contrast with the genetically homogeneous group of German isolates. On the other hand, indistinguishable or closely related genetic profiles among epidemiologically unrelated samples were detected. It was concluded that the high genetic variation was probably a result of a lack of strong selection factors that would influence the evolution of type E. The wide genetic biodiversity observed among type E isolates indicates the value of DNA-based typing methods as a tool in contamination studies in the food industry and in investigations of botulism outbreaks. PMID:10224001

  10. Severity and impact of xerostomia in patients treated with botulinum toxin type b for cervical dystonia: Observations on the quality of life of patients with xerostomia

    OpenAIRE

    Hogan, Patrick; Charles, P. David; Wooten Watts, Maureen; Massey, Janice M.; Miller, Tamara; Mackowiack, John

    2004-01-01

    Background: Although dry mouth (xerostomia) has been reported with botulinum toxin type B used as treatment for cervical dystonia, the impact of this adverse effect (AE) on patients' activities of daily living (ADLs) has not been assessed.

  11. Clostridium botulinum group I strain genotyping by 15-locus multilocus variable-number tandem-repeat analysis

    NARCIS (Netherlands)

    Fillo, S.; Giordani, F.; Anniballi, F.; Gorgé, O.; Ramisse, V.; Vergnaud, G.; Riehm, J.M.; Scholz, H.C.; Splettstoesser, W.D.; Kieboom, J.; Olsen, J.-S.; Fenicia, L.; Lista, F.

    2011-01-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of

  12. Botulinum toxin drugs: brief history and outlook.

    Science.gov (United States)

    Dressler, D

    2016-03-01

    The global botulinum toxin (BT) market is currently undergoing rapid changes: this may be the time to review the history and the future of BT drug development. Since the early 1990s Botox(®) and Dysport(®) dominated the international BT market. Later, Myobloc(®)/NeuroBloc(®), a liquid BT type B drug, came out, but failed. Xeomin(®) is the latest major BT drug. It features removal of complexing proteins and improved neurotoxin purity. Several new BT drugs are coming out of Korea, China and Russia. Scientific challenges for BT drug development include modification of BT's duration of action, its transdermal transport and the design of BT hybrid drugs for specific target tissues. The increased competition will change the global BT market fundamentally and a re-organisation according to large indication groups, such as therapeutic and cosmetic applications, might occur. PMID:26559824

  13. Low pH-Induced Pore Formation by the T Domain of Botulinum Toxin Type A is Dependent upon NaCl Concentration

    Energy Technology Data Exchange (ETDEWEB)

    Lai, B.; Swaminathan, S.; Agarwal, R.; Nelson, L. D.; London, E.

    2010-07-19

    Botulinum neurotoxins (BoNTs) undergo low pH-triggered membrane insertion, resulting in the translocation of their light (catalytic) chains into the cytoplasm. The T (translocation) domain of the BoNT heavy chain is believed to carry out translocation. Here, the behavior of isolated T domain from BoNT type A has been characterized, both in solution and when associated with model membranes. When BoNT T domain prepared in the detergent dodecylmaltoside was diluted into aqueous solution, it exhibited a low pH-dependent conformational change below pH 6. At low pH the T domain associated with, and formed pores within, model membrane vesicles composed of 30 mol% dioleoylphosphatidylglycerol/70 mol% dioleoylphosphatidylcholine. Although T domain interacted with vesicles at low (50 mM) and high (400 mM) NaCl concentrations, the interaction required much less lipid at low salt. However, even at high lipid concentrations pore formation was much more pronounced at low NaCl concentrations than at high NaCl concentration. Increasing salt concentration after insertion in the presence of 50 mM NaCl did not decrease pore formation. A similar effect of NaCl concentration upon pore formation was observed in vesicles composed solely of dioleoylphosphatidylcholine, showing that the effect of NaCl did not solely involve modulation of electrostatic interactions between protein and anionic lipids. These results indicate that some feature of membrane-bound T domain tertiary structure critical for pore formation is highly dependent upon salt concentration.

  14. Crystallization and preliminary X-ray analysis of the HA3 component of Clostridium botulinum type C progenitor toxin

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Toshio; Tonozuka, Takashi; Kotani, Mao; Obata, Kanae [Department of Applied Biological Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509 (Japan); Oguma, Keiji [Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558 (Japan); Nishikawa, Atsushi, E-mail: nishikaw@cc.tuat.ac.jp [Department of Applied Biological Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509 (Japan); CREST, Japan Science and Technology Agency (Japan)

    2007-12-01

    HA3, a 70 kDa haemagglutinating protein, is a precursor form of HA3a and HA3b, the subcomponents of Clostridium botulinum type C 16S progenitor toxin. In this report, recombinant HA3 protein was overexpressed in Escherichia coli, purified and crystallized. HA3, a 70 kDa haemagglutinating protein, is a precursor form of HA3a and HA3b, the subcomponents of Clostridium botulinum type C 16S progenitor toxin. In this report, recombinant HA3 protein was overexpressed in Escherichia coli, purified and crystallized. Diffraction data were collected to 2.6 Å resolution and the crystal belonged to the hexagonal space group P6{sub 3}. Matthews coefficient and self-rotation function calculations indicate that there is probably one molecule of HA3 in the asymmetric unit. A search for heavy-atom derivatives has been undertaken.

  15. Efficacy of Botulinum toxin type A in treatment of different forms of focal dystonias in the Serbian population: Experience of the Botulinum Toxin Outpatients Department

    Directory of Open Access Journals (Sweden)

    Tomić Aleksandra

    2015-01-01

    Full Text Available Background/Aim. Botulinum toxin (BTX irreversibly inhibits presynaptic acetylcholine release with subsequent relaxation of abnormally contracting muscles. It is an effective and well tolerated treatment with long-term benefit in a variety of movement disorders and other neurological and non-neurological disturbances. The aim of our study was to present our experience with BTX type A in treatment of different forms of focal dystonias. Мethods. А hundred of patients with different focal dystonias (spastic torticollis, blepharospasm and graphospasm from the Botulinum Toxin Outpatients Department, Clinic for Neurology, Clinical Center of Serbia, were included in the study. All the patients were examined and rated at baseline visit prior to BTX application and on the following visit, after 3-4 months, using self-assessment improvement questionnaire and standardized rating scales. Results. The improvement of ≥ 50% was presented in 68.2% of all (199 the analyzed applications. Independent predictors of good response to the therapy (improvement ≥ 50% were male sex (p = 0.011, the presence of sensory trick (p = 0.013 and the total number of BTX applications (p = 0.002. The patients with spastic torticollis and blepharospasm showed a statistically significantly better BTX effect (improvement 57.3 ± 27.5% and 54.1 ± 28.3%, respectively than the graphospasm group (26.7 ± 25.6%. Most of the patients did not have therapy complications (81.4% and 72% in two applications. Side effects in the remaining patients (muscle weakness, dysphagia, ptosis, double vision, neck weakness and lacrimal dysfunction lasted for 28.3 ± 18.6 days after the first treatment and 32.5 ± 36.2 days after the second one. Conclusion. BTX is safe and highly effective in long-term treatment of patients with different forms of focal dystonia, with only mild and well-tolerated side-effects. [Projekat Ministarstva nauke Republike Srbije, br. 175090

  16. Clostridium botulinum Group I Strain Genotyping by 15-Locus Multilocus Variable-Number Tandem-Repeat Analysis ▿ †

    Science.gov (United States)

    Fillo, Silvia; Giordani, Francesco; Anniballi, Fabrizio; Gorgé, Olivier; Ramisse, Vincent; Vergnaud, Gilles; Riehm, Julia M.; Scholz, Holger C.; Splettstoesser, Wolf D.; Kieboom, Jasper; Olsen, Jaran-Strand; Fenicia, Lucia; Lista, Florigio

    2011-01-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of C. botulinum isolates of any BoNT type is relevant for both epidemiological studies and forensic microbiology. A 10-locus multiple-locus variable-number tandem-repeat analysis (MLVA) was previously applied to isolates of C. botulinum type A. The present study includes five additional loci designed to better address proteolytic B and F serotypes. We investigated 79 C. botulinum group I strains isolated from human and food samples in several European countries, including types A (28), B (36), AB (4), and F (11) strains, and 5 nontoxic Clostridium sporogenes. Additional data were deduced from in silico analysis of 10 available fully sequenced genomes. This 15-locus MLVA (MLVA-15) scheme identified 86 distinct genotypes that clustered consistently with the results of amplified fragment length polymorphism (AFLP) and MLVA genotyping in previous reports. An MLVA-7 scheme, a subset of the MLVA-15, performed on a lab-on-a-chip device using a nonfluorescent subset of primers, is also proposed as a first-line assay. The phylogenetic grouping obtained with the MLVA-7 does not differ significantly from that generated by the MLVA-15. To our knowledge, this report is the first to analyze genetic variability among all of the C. botulinum group I serotypes by MLVA. Our data provide new insights into the genetic variability of group I C. botulinum isolates worldwide and demonstrate that this group is genetically highly diverse. PMID:22012011

  17. Clostridium botulinum group I strain genotyping by 15-locus multilocus variable-number tandem-repeat analysis.

    Science.gov (United States)

    Fillo, Silvia; Giordani, Francesco; Anniballi, Fabrizio; Gorgé, Olivier; Ramisse, Vincent; Vergnaud, Gilles; Riehm, Julia M; Scholz, Holger C; Splettstoesser, Wolf D; Kieboom, Jasper; Olsen, Jaran-Strand; Fenicia, Lucia; Lista, Florigio

    2011-12-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of C. botulinum isolates of any BoNT type is relevant for both epidemiological studies and forensic microbiology. A 10-locus multiple-locus variable-number tandem-repeat analysis (MLVA) was previously applied to isolates of C. botulinum type A. The present study includes five additional loci designed to better address proteolytic B and F serotypes. We investigated 79 C. botulinum group I strains isolated from human and food samples in several European countries, including types A (28), B (36), AB (4), and F (11) strains, and 5 nontoxic Clostridium sporogenes. Additional data were deduced from in silico analysis of 10 available fully sequenced genomes. This 15-locus MLVA (MLVA-15) scheme identified 86 distinct genotypes that clustered consistently with the results of amplified fragment length polymorphism (AFLP) and MLVA genotyping in previous reports. An MLVA-7 scheme, a subset of the MLVA-15, performed on a lab-on-a-chip device using a nonfluorescent subset of primers, is also proposed as a first-line assay. The phylogenetic grouping obtained with the MLVA-7 does not differ significantly from that generated by the MLVA-15. To our knowledge, this report is the first to analyze genetic variability among all of the C. botulinum group I serotypes by MLVA. Our data provide new insights into the genetic variability of group I C. botulinum isolates worldwide and demonstrate that this group is genetically highly diverse.

  18. Botulinum Toxin Type a Injection, Followed by Home-Based Functional Training for Upper Limb Hemiparesis after Stroke

    Science.gov (United States)

    Takekawa, Toru; Kakuda, Wataru; Taguchi, Kensuke; Ishikawa, Atsushi; Sase, Yousuke; Abo, Masahiro

    2012-01-01

    Botulinum toxin type A (BoNT-A) has been reported to be an effective treatment for limb spasticity after stroke. However, the reduction in the spasticity after BoNT-A injection alone does not ensure an improvement in the active motor function of the affected limb. The aim of this study was to clarify the clinical effects of a BoNT-A injection,…

  19. Botulinum toxin: yesterday, today, tomorrow

    Directory of Open Access Journals (Sweden)

    A. R. Artemenko

    2013-01-01

    Full Text Available Botulinum toxin (BoNT is a bacterial neurotoxin presented with seven serotypes that inhibit neurotransmitter release from nerve endings. The serotypes of BoNT are antigenically dissimilar, act via different, but interconnected mechanisms, and are not interchangeable. The activity of BoNT is associated with impaired neuroexocytosis occurring in several steps: from the binding of BoNT to its specific receptor on the axon terminal membrane to the proteolytic enzymatic cleavage of SNARE substrate. The effect of BoNT is considered to be restricted to the peripheral nervous system, but when given in particularly high doses, it has been recently shown to affect individual brain structures. In addition, by modulating peripheral afferentation, BoNT may influence the excitability of central neuronal structures at both spinal and cortical levels. Only BoNT serotypes A and B are used in clinical practice and aesthetic medicine. The type A has gained the widest acceptance as a therapeutic agent for more than 100 abnormalities manifesting themselves as muscular hyperactivity, hyperfunction of endocrine gland, and chronic pain. The effect of BoNT preparations shows itself 2-5 days after injection, lasts 3 months or more, and gradually decreases with as a result of pharmacokinetic and intracellular reparative processes. Biotechnology advances and potentialities allow purposefully modification of the protein molecular structure of BoNT, which expands the use and efficiency of performed therapy with neurotoxins. Recombinant technologies provide a combination of major therapeutic properties of each used BoNT serotype and expand indications for recombinant chimeric toxins.

  20. Efficacy of Long-term Effect and Repeat Intraarticular Botulinum toxin in Patients with Painful Total Joint Arthroplasty: A Retrospective Study

    OpenAIRE

    Singh, Jasvinder A

    2014-01-01

    Objective Based on recent success of intra-articular (IA) Botulinum neurotoxin type A (BoNT/A; OnabotulinumtoxinA) in patients with osteoarthritis, we examined if repeat IA-BoNT/A is an effective antinociceptive in patients with refractory arthroplasty pain. Methods 11 patients with refractory chronic arthroplasty joint pain without any evidence of infection or prosthesis loosening were referred by orthopedic surgeons. After discussion of off-label use, each patient underwent IA injection of ...

  1. Lumbar Sympathetic Block with Botulinum Toxin Type B for Complex Regional Pain Syndrome: A Case Study.

    Science.gov (United States)

    Choi, Eunjoo; Cho, Chan Woo; Kim, Hye Young; Lee, Pyung Bok; Nahm, Francis Sahngun

    2015-01-01

    Lumbar sympathetic block (LSB) is an effective method for relief of sympathetically mediated pain in the lower extremities. To prolong the sympathetic blockade, sympathetic destruction with alcohol or radiofrequency has been used. The pre-ganglionic sympathetic nerves are cholinergic, and botulinum toxin (BTX) has been found to inhibit the release of acetylcholine at the cholinergic nerve terminals. Moreover, BTX type B (BTX-B) is more convenient to use than BTX type A. Based on these findings, we performed LSB on the 2 patients with complex regional pain syndrome (CRPS) in the lower extremity. Levobupivacaine 0.25% 5 mL mixed with BTX-B 5,000 IU was given under fluoroscopic guidance. Two months after LSB with BTX-B, pain intensity and the Leeds assessment of neuropathic symptoms and signs (LANSS) score were significantly reduced. Allodynia and coldness disappeared and skin color came back to normal. In conclusion, BTX-B can produce an efficacious and durable sympathetic blocking effect on patients with CRPS.

  2. Lumbar Sympathetic Block with Botulinum Toxin Type B for Complex Regional Pain Syndrome: A Case Study.

    Science.gov (United States)

    Choi, Eunjoo; Cho, Chan Woo; Kim, Hye Young; Lee, Pyung Bok; Nahm, Francis Sahngun

    2015-01-01

    Lumbar sympathetic block (LSB) is an effective method for relief of sympathetically mediated pain in the lower extremities. To prolong the sympathetic blockade, sympathetic destruction with alcohol or radiofrequency has been used. The pre-ganglionic sympathetic nerves are cholinergic, and botulinum toxin (BTX) has been found to inhibit the release of acetylcholine at the cholinergic nerve terminals. Moreover, BTX type B (BTX-B) is more convenient to use than BTX type A. Based on these findings, we performed LSB on the 2 patients with complex regional pain syndrome (CRPS) in the lower extremity. Levobupivacaine 0.25% 5 mL mixed with BTX-B 5,000 IU was given under fluoroscopic guidance. Two months after LSB with BTX-B, pain intensity and the Leeds assessment of neuropathic symptoms and signs (LANSS) score were significantly reduced. Allodynia and coldness disappeared and skin color came back to normal. In conclusion, BTX-B can produce an efficacious and durable sympathetic blocking effect on patients with CRPS. PMID:26431145

  3. Association of toxin-producing Clostridium botulinum with the macroalga Cladophora in the Great Lakes.

    Science.gov (United States)

    Chun, Chan Lan; Ochsner, Urs; Byappanahalli, Muruleedhara N; Whitman, Richard L; Tepp, William H; Lin, Guangyun; Johnson, Eric A; Peller, Julie; Sadowsky, Michael J

    2013-03-19

    Avian botulism, a paralytic disease of birds, often occurs on a yearly cycle and is increasingly becoming more common in the Great Lakes. Outbreaks are caused by bird ingestion of neurotoxins produced by Clostridium botulinum, a spore-forming, gram-positive, anaerobe. The nuisance, macrophytic, green alga Cladophora (Chlorophyta; mostly Cladophora glomerata L.) is a potential habitat for the growth of C. botulinum. A high incidence of botulism in shoreline birds at Sleeping Bear Dunes National Lakeshore (SLBE) in Lake Michigan coincides with increasingly massive accumulations of Cladophora in nearshore waters. In this study, free-floating algal mats were collected from SLBE and other shorelines of the Great Lakes between June and October 2011. The abundance of C. botulinum in algal mats was quantified and the type of botulism neurotoxin (bont) genes associated with this organism were determined by using most-probable-number PCR (MPN-PCR) and five distinct bont gene-specific primers (A, B, C, E, and F). The MPN-PCR results showed that 16 of 22 (73%) algal mats from the SLBE and 23 of 31(74%) algal mats from other shorelines of the Great Lakes contained the bont type E (bont/E) gene. C. botulinum was present up to 15000 MPN per gram dried algae based on gene copies of bont/E. In addition, genes for bont/A and bont/B, which are commonly associated with human diseases, were detected in a few algal samples. Moreover, C. botulinum was present as vegetative cells rather than as dormant spores in Cladophora mats. Mouse toxin assays done using supernatants from enrichment of Cladophora containing high densities of C. botulinum (>1000 MPN/g dried algae) showed that Cladophora-borne C. botulinum were toxin-producing species (BoNT/E). Our results indicate that Cladophora provides a habitat for C. botulinum, warranting additional studies to better understand the relationship between this bacterium and the alga, and how this interaction potentially contributes to botulism

  4. Association of toxin-producing Clostridium botulinum with the macroalga Cladophora in the Great Lakes

    Science.gov (United States)

    Chun, Chan Lan; Ochsner, Urs; Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Tepp, William H.; Lin, Guangyun; Johnson, Eric A.; Peller, Julie; Sadowsky, Michael J.

    2013-01-01

    Avian botulism, a paralytic disease of birds, often occurs on a yearly cycle and is increasingly becoming more common in the Great Lakes. Outbreaks are caused by bird ingestion of neurotoxins produced by Clostridium botulinum, a spore-forming, gram-positive, anaerobe. The nuisance, macrophytic, green alga Cladophora (Chlorophyta; mostly Cladophora glomerata L.) is a potential habitat for the growth of C. botulinum. A high incidence of botulism in shoreline birds at Sleeping Bear Dunes National Lakeshore (SLBE) in Lake Michigan coincides with increasingly massive accumulations of Cladophora in nearshore waters. In this study, free-floating algal mats were collected from SLBE and other shorelines of the Great Lakes between June and October 2011. The abundance of C. botulinum in algal mats was quantified and the type of botulism neurotoxin (bont) genes associated with this organism were determined by using most-probable-number PCR (MPN-PCR) and five distinct bont gene-specific primers (A, B, C, E, and F). The MPN-PCR results showed that 16 of 22 (73%) algal mats from the SLBE and 23 of 31(74%) algal mats from other shorelines of the Great Lakes contained the bont type E (bont/E) gene. C. botulinum was present up to 15 000 MPN per gram dried algae based on gene copies of bont/E. In addition, genes for bont/A and bont/B, which are commonly associated with human diseases, were detected in a few algal samples. Moreover, C. botulinum was present as vegetative cells rather than as dormant spores in Cladophora mats. Mouse toxin assays done using supernatants from enrichment of Cladophora containing high densities of C. botulinum (>1000 MPN/g dried algae) showed that Cladophora-borne C. botulinum were toxin-producing species (BoNT/E). Our results indicate that Cladophora provides a habitat for C. botulinum, warranting additional studies to better understand the relationship between this bacterium and the alga, and how this interaction potentially contributes to botulism

  5. THE PROPERTIES AND LONGITUDINAL EXPERIENCE OF CHINESE TYPE A BOTULINUM TOXIN FOR THE TREATMENT OF FOCAL DYSTONIA AND HEMIFACIAL SPASM

    Institute of Scientific and Technical Information of China (English)

    万新华; 汤晓芙; 王荫椿

    2003-01-01

    Objective. To introduce the properties of Chinese type A botulinum toxin (CBTXA, made by Lanzhou Institute of Biological Products), and its long-term effect for focal dystonia and hemifacial spasm. Method. The purity and recovery of crude and crystalline toxin were tested. Long-term data from 305 patients with hemifacial spasm (HFS), blepharospasm (BS) and cervical dystonia (CD) were evaluated and subgroups of patients received CBTXA injections between 1994 and 2000 in at least six separate treatment sessions, with follow up for 2-8 years. The therapeutic results of the last session CBTXA injections were analyzed in comparison with the first session. Result. CBTXA purity was high[(2.55~2.60)×107D50/mgPr, A260/A280 ≤0.55, high molecular substance accounted for 99.2% of total proteins]. Long term treatment with CBTXA in patients with focal dystonia and HFS was not associated with any decline in benefit, and efficacy may improve slightly with repeat treatments. CBTXA is an excellent long-term treatment of HFS, BS and CD. Conchusion. We conclude that Chinese type A botulinum toxin is of botulinum toxin therapy quality standard according to results obtained from the basic study and long-term clinical applications. The reinjection of CBTXA significantly improves the quality of life of most patients and is a safe, effective and comparatively economical treatment for patients with focal dystonia and HFS.

  6. Detection of Clostridium botulinum in liquid manure and biogas plant wastes.

    Science.gov (United States)

    Neuhaus, Jürgen; Schrödl, Wieland; Shehata, Awad A; Krüger, Monika

    2015-09-01

    Biogas plants have been considered as a source for possible amplification and distribution of pathogenic bacteria capable of causing severe infections in humans and animals. Manure and biogas wastes could be sources for spore-forming bacteria such as Clostridium botulinum. In the present study, 24 liquid manure and 84 biogas waste samples from dairies where the majority of the cows suffered from chronic botulism were investigated for the presence of botulinum neurotoxins (BoNT) and C. botulinum spores. The prevalence of BoNT/A, B, C, D, and E in biogas wastes was 16.6, 8.3, 10.7, 7.1, and 10.8 %, respectively, while in manure, the prevalence was 0.0, 0.0, 0.0, 8.3, and 4.1 %, respectively. After enrichment of samples in reinforced cultural medium, they were tested for C. botulinum BoNT/A, B, C, D, and E using ELISA (indirect C. botulinum detection). The prevalence of C. botulinum type A, B, C, D, and E samples in biogas wastes was 20.2, 15.5, 19, 10.7, and 34.8 %, respectively, while the prevalence in liquid manure was 0.0, 0.0, 0.0, 8.3, and 12.5 %, respectively. In conclusion, the occurrence of BoNT and C. botulinum spores in biogas waste of diseased animals indicates an increased and underestimated hygienic risk. Application of digestates from biogas fermentations as fertilizers could lead to an accumulation of long lifespan spores in the environment and could be a possible health hazard. PMID:25753763

  7. Detection of Clostridium botulinum in liquid manure and biogas plant wastes.

    Science.gov (United States)

    Neuhaus, Jürgen; Schrödl, Wieland; Shehata, Awad A; Krüger, Monika

    2015-09-01

    Biogas plants have been considered as a source for possible amplification and distribution of pathogenic bacteria capable of causing severe infections in humans and animals. Manure and biogas wastes could be sources for spore-forming bacteria such as Clostridium botulinum. In the present study, 24 liquid manure and 84 biogas waste samples from dairies where the majority of the cows suffered from chronic botulism were investigated for the presence of botulinum neurotoxins (BoNT) and C. botulinum spores. The prevalence of BoNT/A, B, C, D, and E in biogas wastes was 16.6, 8.3, 10.7, 7.1, and 10.8 %, respectively, while in manure, the prevalence was 0.0, 0.0, 0.0, 8.3, and 4.1 %, respectively. After enrichment of samples in reinforced cultural medium, they were tested for C. botulinum BoNT/A, B, C, D, and E using ELISA (indirect C. botulinum detection). The prevalence of C. botulinum type A, B, C, D, and E samples in biogas wastes was 20.2, 15.5, 19, 10.7, and 34.8 %, respectively, while the prevalence in liquid manure was 0.0, 0.0, 0.0, 8.3, and 12.5 %, respectively. In conclusion, the occurrence of BoNT and C. botulinum spores in biogas waste of diseased animals indicates an increased and underestimated hygienic risk. Application of digestates from biogas fermentations as fertilizers could lead to an accumulation of long lifespan spores in the environment and could be a possible health hazard.

  8. SjAPI-2 is the first member of a new neurotoxin family with Ascaris-type fold and KCNQ1 inhibitory activity.

    Science.gov (United States)

    Chen, Jing; Zhang, Chuangeng; Yang, Weishan; Cao, Zhijian; Li, Wenxin; Chen, Zongyun; Wu, Yingliang

    2015-08-01

    Peptides with Ascaris-type fold are a new kind of toxins founded from venomous animals recently. Functionally, these unique toxin peptides had been identified as potent protease inhibitors, which was similar to other known Ascaris-type peptides from non-venomous animals. Whether Ascaris-type peptides from venom animals have neurotoxin activities remains unclear. Here, a scorpion toxin SjAPI-2 with Ascaris-type fold was characterized to have a neurotoxin activity, which can selectively inhibit KCNQ1 potassium channel. SjAPI-2 had 62 amino acid residues, including 10 cysteine residues. Charged residue analyses showed that two acidic residues of SjAPI-2 were regionally distributed, and 10 basic residues were distributed widely throughout the whole peptide, which was similar to classical potassium channel toxins. Pharmacological studies confirmed that SjAPI-2 was a selective KCNQ1 potassium channel inhibitor with weak effects on other potassium channels, such as Kv1.1, Kv1.2, Kv1.3, SKCa2, SKCa3, and IKCa channels. Concentration-dependent studies showed that SjAPI-2 inhibited the KCNQ1 potassium channel with an IC50 of 771.5±169.9 nM. To the best of our knowledge, SjAPI-2 is the first neurotoxin with a unique Ascaris-type fold, providing novel insights into the divergent evolution of neurotoxins from venomous animals. PMID:26014142

  9. CLINICAL APPLICATION OF BOTULINUM TOXIN TYPE B IN MOVEMENT DISORDERS AND AUTONOMIC SYMPTOMS

    Institute of Scientific and Technical Information of China (English)

    Xin-hua Wan; Kevin Dat Vuong; Joseph Jankovic

    2005-01-01

    Objective To evaluate efficacy and safety of botulinum toxin type B (BTX-B) in treatment of movement disorders including blepharospasm, oromandibular dystonia, hemifacial spasm, tremor, tics, and hypersecretory disorders such as sialorrhea and hyperhidrosis.Methods A retrospective study of BTX-B injections in treatment of 58 patients with various neurological disorders was performed. The mean follow-up time was 0.9 ± 0.8 years. Results of the first and last treatment of patients with at least 3injection sessions were compared.Results The response of 58 patients to a total of 157 BTX-B treatment sessions was analyzed. Of the 157 treatment sessions, 120 sessions (76.4%) resulted in moderate or marked improvement while 17 sessions (10.8%) had no response.The clinical benefits after BTX-B treatment lasted an average of 14 weeks. Of the 41 patients with at least 3 injection sessions (mean 10 ± 8.6), most patients needed increased dosage upon the last session compared to the first session. Nineteen patients (32.8%) with 27 sessions (17.2%) reported adverse effects with BTX-B treatment.Conclusios Though most patients require increased dosage to maintain effective response after repeated injections,BTX-B is an effective and safe treatment drug for a variety of movement disorders, as well as drooling and hyperhidrosis.

  10. Ultrasound-Guided Nerve Block with Botulinum Toxin Type A for Intractable Neuropathic Pain.

    Science.gov (United States)

    Moon, Young Eun; Choi, Jung Hyun; Park, Hue Jung; Park, Ji Hye; Kim, Ji Hyun

    2016-01-01

    Neuropathic pain includes postherpetic neuralgia (PHN), painful diabetic neuropathy (PDN), and trigeminal neuralgia, and so on. Although various drugs have been tried to treat neuropathic pain, the effectiveness of the drugs sometimes may be limited for chronic intractable neuropathic pain, especially when they cannot be used at an adequate dose, due to undesirable severe side effects and the underlying disease itself. Botulinum toxin type A (BoNT-A) has been known for its analgesic effect in various pain conditions. Nevertheless, there are no data of nerve block in PHN and PDN. Here, we report two patients successfully treated with ultrasound-guided peripheral nerve block using BoNT-A for intractable PHN and PDN. One patient had PHN on the left upper extremity and the other patient had PDN on a lower extremity. Due to side effects of drugs, escalation of the drug dose could not be made. We injected 50 Botox units (BOTOX(®), Allergan Inc., Irvine, CA, USA) into brachial plexus and lumbar plexus, respectively, under ultrasound. Their pain was significantly decreased for about 4-5 months. Ultrasound-guided nerve block with BoNT-A may be an effective analgesic modality in a chronic intractable neuropathic pain especially when conventional treatment failed to achieve adequate pain relief. PMID:26761032

  11. Ultrasound-Guided Nerve Block with Botulinum Toxin Type A for Intractable Neuropathic Pain

    Directory of Open Access Journals (Sweden)

    Young Eun Moon

    2016-01-01

    Full Text Available Neuropathic pain includes postherpetic neuralgia (PHN, painful diabetic neuropathy (PDN, and trigeminal neuralgia, and so on. Although various drugs have been tried to treat neuropathic pain, the effectiveness of the drugs sometimes may be limited for chronic intractable neuropathic pain, especially when they cannot be used at an adequate dose, due to undesirable severe side effects and the underlying disease itself. Botulinum toxin type A (BoNT-A has been known for its analgesic effect in various pain conditions. Nevertheless, there are no data of nerve block in PHN and PDN. Here, we report two patients successfully treated with ultrasound-guided peripheral nerve block using BoNT-A for intractable PHN and PDN. One patient had PHN on the left upper extremity and the other patient had PDN on a lower extremity. Due to side effects of drugs, escalation of the drug dose could not be made. We injected 50 Botox units (BOTOX®, Allergan Inc., Irvine, CA, USA into brachial plexus and lumbar plexus, respectively, under ultrasound. Their pain was significantly decreased for about 4–5 months. Ultrasound-guided nerve block with BoNT-A may be an effective analgesic modality in a chronic intractable neuropathic pain especially when conventional treatment failed to achieve adequate pain relief.

  12. Antinociceptive Effects of Botulinum Toxin Type A on Trigeminal Neuropathic Pain.

    Science.gov (United States)

    Yang, K Y; Kim, M J; Ju, J S; Park, S K; Lee, C G; Kim, S T; Bae, Y C; Ahn, D K

    2016-09-01

    Previous studies have demonstrated that botulinum toxin type A (BoNT-A) attenuates orofacial nociception. However, there has been no evidence of the participation of the voltage-gated sodium channels (Navs) in the antinociceptive mechanisms of BoNT-A. This study investigated the cellular mechanisms underlying the antinociceptive effects of BoNT-A in a male Sprague-Dawley rat model of trigeminal neuropathic pain produced by malpositioned dental implants. The left mandibular second molar was extracted under anesthesia, followed by a miniature dental implant placement to induce injury to the inferior alveolar nerve. Mechanical allodynia was monitored after subcutaneous injection of BoNT-A at 3, 7, or 12 d after malpositioned dental implant surgery. Subcutaneous injections of 1 or 3 U/kg of BoNT-A on postoperative day 3 significantly attenuated mechanical allodynia, although 0.3 U/kg of BoNT-A did not affect the air-puff threshold. A single injection of 3 U/kg of BoNT-A produced prolonged antiallodynic effects over the entire experimental period. Treatment with BoNT-A on postoperative days 7 and 12, when pain had already been established, also produced prolonged antiallodynic effects. Double treatments with 1 U/kg of BoNT-A produced prolonged, more antiallodynic effects as compared with single treatments. Subcutaneous administration of 3 U/kg of BoNT-A significantly inhibited the upregulation of Nav isoform 1.7 (Nav1.7) expression in the trigeminal ganglion in the nerve-injured animals. These results suggest that antinociceptive effects of BoNT-A are mediated by an inhibition of upregulated Nav1.7 expression in the trigeminal ganglion. BoNT-A is therefore a potential new therapeutic agent for chronic pain control, including neuropathic pain. PMID:27418174

  13. Botulinum toxin type A for neuropathic pain in patients with spinal cord injury

    Science.gov (United States)

    Han, Zee‐A; Song, Dae Heon; Oh, Hyun‐Mi

    2016-01-01

    Objective To evaluate the analgesic effect of botulinum toxin type A (BTX‐A) on patients with spinal cord injury‐associated neuropathic pain. Methods The effect of BTX‐A on 40 patients with spinal cord injury‐associated neuropathic pain was investigated using a randomized, double‐blind, placebo‐controlled design. A 1‐time subcutaneous BTX‐A (200U) injection was administered to the painful area. Visual analogue scale (VAS) scores (0–100mm), the Korean version of the short‐form McGill Pain Questionnaire, and the World Health Organization WHOQOL‐BREF quality of life assessment were evaluated prior to treatment and at 4 and 8 weeks after the injection. Results At 4 and 8 weeks after injection, the VAS score for pain was significantly reduced by 18.6 ± 16.8 and 21.3 ± 26.8, respectively, in the BTX‐A group, whereas it was reduced by 2.6 ± 14.6 and 0.3 ± 19.5, respectively, in the placebo group. The pain relief was associated with preservation of motor or sensory function below the neurological level of injury. Among the responders in the BTX‐A group, 55% and 45% reported pain relief of 20% or greater at 4 and 8 weeks, respectively, after the injection, whereas only 15% and 10% of the responders in the placebo group reported a similar level of pain relief. Improvements in the score for the physical health domain of the WHOQOL‐BREF in the BTX‐A group showed a marginal trend toward significance (p = 0.0521) at 4 weeks after the injection. Interpretation These results indicate that BTX‐A may reduce intractable chronic neuropathic pain in patients with spinal cord injury. Ann Neurol 2016;79:569–578 PMID:26814620

  14. Kinetic and Reaction Pathway Analysis in the Application of Botulinum Toxin A for Wound Healing

    Directory of Open Access Journals (Sweden)

    Frank J. Lebeda

    2012-01-01

    Full Text Available A relatively new approach in the treatment of specific wounds in animal models and in patients with type A botulinum toxin is the focus of this paper. The indications or conditions include traumatic wounds (experimental and clinical, surgical (incision wounds, and wounds such as fissures and ulcers that are signs/symptoms of disease or other processes. An objective was to conduct systematic literature searches and take note of the reactions involved in the healing process and identify corresponding pharmacokinetic data. From several case reports, we developed a qualitative model of how botulinum toxin disrupts the vicious cycle of muscle spasm, pain, inflammation, decreased blood flow, and ischemia. We transformed this model into a minimal kinetic scheme for healing chronic wounds. The model helped us to estimate the rate of decline of this toxin's therapeutic effect by calculating the rate of recurrence of clinical symptoms after a wound-healing treatment with this neurotoxin.

  15. Effects of potassium sorbate and other antibotulinal agents on germination and outgrowth of Clostridium botulinum type E spores in microcultures.

    OpenAIRE

    Seward, R A; Deibel, R. H.; Lindsay, R C

    1982-01-01

    The effects of potassium sorbate, sodium hypophosphite, sodium tripolyphosphate, sodium nitrite, and linoleic acid on the germination and outgrowth of Clostridium botulinum type E spores were studied in microcultures. At pH 5.8 to 6.0 in liver veal agar, the germination rate was decreased to nearly zero with 1.0, 1.5, or 2.0% sorbate. At pH 7.0 t 7.2, these levels of sorbate afforded germination and outgrowth of abnormally shaped cells that were defective in cell division. At the high pH rang...

  16. Botulinum toxin type A normalizes alterations in urothelial ATP and NO release induced by chronic spinal cord injury

    OpenAIRE

    Smith, Christopher P.; Gangitano, David A; Munoz, Alvaro; Salas, Nilson A.; Boone, Timothy B.; Aoki, K Roger; Francis, Joseph; Somogyi, George T.

    2007-01-01

    The purpose of this paper was to simultaneously examine changes in urothelial ATP and NO release in normal and spinal cord injured animals as well as in spinal cord injured animals treated with botulinum toxin type A (BoNT-A). Furthermore we correlated changes in transmitter release with functional changes in bladder contraction frequency, and determined the effects of BoNT-A on bladder efferent nerve function. Normal and spinal cord injured rat bladders were injected on day 0 with either veh...

  17. Botulinum toxin type-A effect as a preemptive treatment in a model of acute trigeminal pain: a pre-clinical double-blind and placebo-controlled study

    Directory of Open Access Journals (Sweden)

    Elcio Juliato Piovesan

    2011-02-01

    Full Text Available The purpose of this study was to investigate if botulinum neurotoxin type-A (BoNT/A had a preemptive antinociceptive effect in a formalin-induced orofacial pain model (FT. To test this hypothesis, male Rattus norvegicus were injected with isotonic saline solution 0.9% or BoNT/A administered as a 40 μl bolus, lateral to their nose, at 24 hours, 8, 15, 22, 29 or 36 days pre-FT. The procedures were repeated 42 days later. Influence on motor activity was assessed through the open-field test. Pain scores corresponded to the time spent rubbing and flicking the injected area. Animals pre-treated with BoNT/A at the first protocol (8 days subgroup showed reduced inflammatory scores (p=0.011. For the other groups no significant results were observed at any phase. Motor activity was similar in both groups. BoNT/A showed to be effective preventing inflammatory pain up to eight days after the first treatment, an effect not reproduced on the second dose administration.

  18. Inhibition of growth of nonproteolytic Clostridium botulinum type B in sous vide cooked meat products is achieved by using thermal processing but not nisin.

    Science.gov (United States)

    Lindström, M; Mokkila, M; Skyttä, E; Hyytiä-Trees, E; Lähteenmäki, L; Hielm, S; Ahvenainen, R; Korkeala, H

    2001-06-01

    The safety of refrigerated processed foods of extended durability (REPFEDs) with respect to nonproteolytic Clostridium botulinum is under continuous evaluation. In the present study, mild (P7.0(85.0) values 0 to 2 min [P, pasteurization value; z-value 7.0 degrees C; reference temperature 85.0 degrees C]) and increased (P7.0(85.0) values 67 to 515 min) heat treatments were evaluated in relation to survival of nonproteolytic C. botulinum type B spores in sous vide processed ground beef and pork cubes. The use of two concentrations of nisin in inhibition of growth and toxin production by nonproteolytic C. botulinum in the same products was also evaluated. A total of 96 samples were heat processed and analyzed for C. botulinum by BoNT/B gene-specific polmerase chain reaction and for botulinum toxin by a mouse bioassay after storage of 14 to 28 days at 4 and 8 degrees C. Predictably, after mild processing all samples of both products showed botulinal growth, and one ground beef sample became toxic at 8 degrees C. The increased heat processing, equivalent to 67 min at 85 degrees C. resulted in growth but not toxin production of C. botulinum in one ground beef sample in 21 days at 8 degrees C: in the pork cube samples no growth was detected. The increased heating of both products resulted in higher sensory quality than the milder heat treatment. Nisin did not inhibit the growth of nonproteolytic C. botulinum in either product; growth was detected in both products at 4 and 8 degrees C, and ground beef became toxic with all nisin levels within 21 to 28 days at 8 degrees C. Aerobic and lactic acid bacterial counts were reduced by the addition of nisin at 4 degrees C. The study demonstrates that the mild processing temperatures commonly employed in sous vide technology do not eliminate nonproteolytic C. botulinum type B spores. The intensity of each heat treatment needs to be carefully evaluated individually for each product to ensure product safety in relation to

  19. Antibody-induced secondary treatment failure in a patient treated with botulinum toxin type A for glabellar frown lines

    Directory of Open Access Journals (Sweden)

    Stengel G

    2011-11-01

    Full Text Available Gabriele Stengel, Eva Kristina Bee Hautarztpraxis Stengel and Bee, Münster, Germany Abstract: Botulinum toxin type A (BTX-A preparations are widely used nonsurgical treatments for facial wrinkles. Higher doses of BTX-A are also used for therapeutic purposes in the treatment of conditions involving increased muscle tone, such as cervical dystonia. The phenomenon of antibody-induced treatment failure is well known in the therapeutic setting, but reports are also emerging following cosmetic use of BTX-A. We describe the case of a 41-year-old female nurse who developed secondary treatment failure during 6 years of BTX-A treatment for glabellar lines. After a good response to the first BTX-A injection, the intensity and duration of effect decreased after subsequent treatments. Antibody tests revealed a high titer of neutralizing anti-BTX-A antibodies. This case shows secondary treatment failure due to the production of neutralizing antibodies following administration of BTX-A formulations for cosmetic purposes and demonstrates that immunogenicity of BTX-A preparations is an important consideration, even in the cosmetic setting. Keywords: botulinum toxin type A, neutralizing antibodies, antibody-induced treatment failure

  20. Functional effects of botulinum toxin type-A treatment and subsequent stretching of spastic calf muscles: a study in patients with hereditary spastic paraplegia

    NARCIS (Netherlands)

    Niet, M. de; Bot, S.T. de; Warrenburg, B.P.C. van de; Weerdesteijn, V.G.M.; Geurts, A.C.H.

    2015-01-01

    OBJECTIVE: Although calf muscle spasticity is often treated with botulinum toxin type-A, the effects on balance and gait are ambiguous. Hereditary spastic paraplegia is characterized by progressive spasticity and relatively mild muscle weakness of the lower limbs. It is therefore a good model to eva

  1. The Profile of Patients and Current Practice of Treatment of Upper Limb Muscle Spasticity with Botulinum Toxin Type A: An International Survey

    Science.gov (United States)

    Bakheit, Abdel Magid

    2010-01-01

    To document the current practice in relation with the treatment of patients with upper limb spasticity with botulinum toxin type A to inform future research in this area. We designed an international, cross-sectional, noninterventional survey of current practice. Nine hundred and seventy-four patients from 122 investigational centres in 31…

  2. The combined effect of lower-limb multilevel botulinum toxin type A and comprehensive rehabilitation on mobility in children with cerebral palsy : A randomized clinical trial

    NARCIS (Netherlands)

    Scholtes, Vanessa A.; Dallmeijer, Annet J.; Knol, Dirk L.; Speth, Lucianne A.; Maathuis, Carel G.; Jongerius, Peter H.; Becher, Jules G.

    2006-01-01

    Objective: To evaluate the combined effect on mobility of treatment with multilevel botulinum toxin type A (BTX-A) and comprehensive rehabilitation in children with cerebral palsy (CP). Design: Randomized clinical trial using a multiple baseline design. The intervention group was treated 6 weeks aft

  3. The combined effect of lower-limb multilevel botulinum toxin type A and comprehensive rehabilitation on mobility in children with cerebral palsy: A randomized clinical trial

    NARCIS (Netherlands)

    Scholtes, V.A.; Dallmeijer, A.J.; Knol, D.L.; Speth, L.A.; Maathuis, C.G.; Jongerius, P.H.; Becher, J.G.

    2006-01-01

    Objective: To evaluate the combined effect on mobility of treatment with multilevel botulinum toxin type A (BTX-A) and comprehensive rehabilitation in children with cerebral palsy (CP). Design: Randomized clinical trial using a multiple baseline design. The intervention group was treated 6 weeks aft

  4. Focal hyperhidrosis secondary to eccrine naevus successfully treated with botulinum toxin type A.

    Science.gov (United States)

    Lera, M; España, A; Idoate, M Á

    2015-08-01

    Eccrine naevus (EN) is a rare skin hamartoma included in the organoid group of epidermal naevi, histologically defined as focal hyperplasia and/or hypertrophy of eccrine glands. Clinically, EN usually presents as hyperhidrotic patches with no visible skin changes, frequently located on the forearms. The decision to treat EN or not usually depends on the grade of hyperhidrosis, but there is no therapeutic consensus because of the rarity of this condition. We present a case diagnosed as EN in an adult patient with severe localized hyperhidrosis, which was successfully treated with botulinum toxin. PMID:25816711

  5. Centrifugal microfluidic platform for ultrasensitive detection of Botulinum Toxin

    Science.gov (United States)

    Botulinum neurotoxin – a global public health threat and category A bioterrorism agent - is the most toxic substance known and one of the most challenging toxins to detect due to its lethality at extremely low concentrations. Hence the live-mouse bioassay because of its superior sensitivity, remains...

  6. A case report of the beneficial effects of botulinum toxin type A on Raynaud phenomenon in a patient with lung cancer

    Science.gov (United States)

    Wang, Lu; Lei, Qi-song; Liu, Yu-ying; Song, Guan-jie; Song, Chun-ling

    2016-01-01

    Abstract Objective: Raynaud phenomenon is a vasospastic disorder affecting the hands and feet, and the efficacies of traditional treatments, such as pharmacological therapies and sympathectomy, are not uniform. Patients with paraneoplastic Raynaud phenomenon do not benefit from the traditional treatments. The use of botulinum toxin type A (BTX-A) for Raynaud phenomenon has been reported for several years; however, there are few reports regarding botulinum toxin type A in the treatment of paraneoplastic Raynaud phenomenon. We describe a case report of the beneficial effects of botulinum toxin type A on Raynaud phenomenon in a patient with lung cancer. Methods: A 63-year-old male complained of pain and discoloration of his fingers and indicated that oral nifedipine and low-dose aspirin were not effective. After approximately 8 months, he was diagnosed with lung cancer. Chemotherapy partially reduced the pain and discoloration of his fingers; however, no significant changes occurred in his fingers after the fourth cycle. We used BTX-A to treat this patient with paraneoplastic RP. A visual analogue scale (VAS) was used to assess the clinical response. Results: After approximately 2 months, the patient reported relief from pain, stiffness, numbness, and cold sensation. Furthermore, no local or general adverse effects were exhibited by the patient. Conclusion: This study used botulinum toxin type A for a patient with paraneoplastic Raynaud phenomenon. Botulinum toxin type A significantly improved the patient's clinical symptoms without significant complications. These findings suggest that BTX-A may represent a good option for the treatment of paraneoplastic RP. PMID:27749585

  7. Inhibition of toxinogenesis of type a Clostridium botulinum in beef using combined treatments

    International Nuclear Information System (INIS)

    A shelf stable beef product was developed on the basis of combined treatments involving a reduction in water activity and using specific anti-microbial agents, thermal processing, vacuum packaging and irradiation. Beef foreshanks were cured, thermally processed and irradiated at 7.5 and 15 kGy. Sensory analysis and microbiological, chemical and lipid stability studies were carried out on the non-inoculated samples. In addition, challenge studies involving samples inoculated with 103 or 105 spores of Clostridium botulinum/g were performed. The samples challenged with 103 spores of C. botulinum and irradiated with 15 kGy did not show toxin production during a storage time of 8 months at 28 deg. C. The non-challenged samples showed low 2-thiobarbituric acid numbers and a significant decrease in residual nitrite during storage. Sensory studies carried out by a trained panel indicated that these samples developed some mild off-flavours, which diminished as the storage time increased, and showed good overall acceptance. Studies to introduce a further safety factor, by adding sodium propionate, are currently in progress. (author)

  8. Treating glabellar lines with botulinum toxin type A-hemagglutinin complex: A review of the science, the clinical data, and patient satisfaction

    Directory of Open Access Journals (Sweden)

    Koenraad De Boulle

    2010-04-01

    Full Text Available Koenraad De Boulle1, Steven Fagien2, Boris Sommer3, Richard Glogau41Aalst Dermatology Clinic, Aalst, Belgium; 2Aesthetic Eyelid Plastic, Surgery, Boca Raton, FL, USA; 3Aesthetic Dermatology, Frankfurt, Germany; 4University of California, San Francisco, USAAbstract: Botulinum toxin type A treatment is the foundation of minimally invasive aesthetic facial procedures. Clinicians and their patients recognize the important role, both negative and positive, that facial expression, particularly the glabellar frown lines, plays in self-perception, emotional well-being, and perception by others. This article provides up-to-date information on fundamental properties and mechanisms of action of the major approved formulations of botulinum toxin type A, summarizes recent changes in naming conventions (nonproprietary names mandated by the United States Food and Drug Administration, and describes the reasons for these changes. The request for these changes provides recognition that formulations of botulinum toxins (eg, onabotulinumtoxinA and abobotulinumtoxinA are not interchangeable and that dosing recommendations cannot be based on any one single conversion ratio. The extensive safety, tolerability, and efficacy data are summarized in detail, including the patient-reported outcomes that contribute to overall patient satisfaction and probability treatment continuation. Based on this in-depth review, the authors conclude that botulinum toxin type A treatment remains a cornerstone of facial aesthetic treatments, and clinicians must realize that techniques and dosing from one formulation cannot be applied to others, that each patient should undergo a full aesthetic evaluation, and that products and procedures must be selected in the context of individual needs and goals.Keywords: onabotulinumtoxinA, botulinum toxin type A, cosmetic, glabellar, patient satisfaction

  9. An aptamer beacon responsive to botulinum toxins.

    Science.gov (United States)

    Bruno, John G; Richarte, Alicia M; Carrillo, Maria P; Edge, Allison

    2012-01-15

    Sixty candidate DNA aptamers were developed against botulinum neurotoxin (BoNT) type A light chain (LC) from ten rounds of selection, resulting in several identical sequences. Secondary structures of the identical aptamers were compared to structures of previously reported BoNT A DNA aptamers. A series of ten candidate loop structures were selected from this comparison as potential binding pockets and aptamer beacons. These candidate beacons were synthesized with 5'-TYE 665 and 3'-Iowa Black quencher labels for comparison of fluorescence levels as a function of BoNT A LC concentration. Only three of the ten candidates exhibited any fluorescence response to increasing levels of BoNT A LC. However, of the two most responsive candidates, one represented a subset loop of the larger more intensely fluorescent double-looped structure, designated Beacon 10. This beacon yielded a lower limit of detection of 1 ng/mL in buffer using a spectrofluorometer and a portable handheld fluorometer, but also responded substantially to BoNT A, B, E holotoxins and heavy or light chain components even in a dilute soil suspension, but not in 50% human serum. Beacon 10 did not respond strongly to a variety of other divergent peptides, suggesting that it is relatively specific to the level of botulinum toxins and is only useful for environmental testing. Beacon 10 also shared short sequence segments with other published BoNT aptamer DNA sequences, suggesting that these may be points of physical contact between the aptamers and BoNTs.

  10. An in vivo analysis of facial muscle change treated with botulinum toxin type A using digital image speckle correlation

    Science.gov (United States)

    Xu, Yan; Palmaccio, Samantha Palmaccio; Bui, Duc; Dagum, Alexander; Rafailovich, Miriam

    Been famous for clinical use from early 1980s, the neuromuscular blocking agent Botulinum toxin type A (BTX-A), has been used to reduce wrinkles for a long time. Only little research has been done to quantify the change of muscle contraction before and after injection and most research paper depend on subjective evaluation from both patients and surgeons. In our research, Digital Image Speckle Correlation (DISC) was employed to study the mechanical properties of skin, contraction mode of muscles (injected) and reaction of neighbor muscle group (un-injected).At the same time, displacement patterns (vector maps)generated by DISC can predict injection locus for surgeons who normally handle it depending only on visual observation.

  11. Effects of botulinum toxin type A for spastic foot in post-stroke patients enrolled in a rehabilitation program

    Directory of Open Access Journals (Sweden)

    Leonardo Halley Carvalho Pimentel

    2014-01-01

    Full Text Available The objective of this study was to evaluate the effects of botulinum toxin type A (BTX-A on spastic foot in stroke patients in a rehabilitation program. Method: Hemiparetic stroke patients (n=21 enrolled in a rehabilitation program were divided into two groups. The first group (n=11 received a total of 300UI BTX-A, and the second group (n=10 received 100 UI BTX-A. All patients were assessed at baseline and 2, 4, 8 and 12 weeks after injection for Modified Ashworth Score, time walking 10 meters, and the Functional Independence Measure (mFIM motor score. Results: The higher-dose group exhibited a significant improvement in spasticity, and both groups showed an improvement in time walking 10 meters and mFIM, with no significant differences between them. Conclusions: Our findings suggest that gains in gait velocity and functional independence were not correlated to BTX-A dose.

  12. Botox (Botulinum Toxin)

    Science.gov (United States)

    ... and rashes clinical tools newsletter | contact Share | Botox (Botulinum Toxin) A A A BEFORE: Crow's feet before Botox ... wrinkles. One such procedure involves the use of botulinum toxin injections. Botulinum toxin is produced by the fermentation ...

  13. Beneficial effects of botulinum toxin type A in trigeminal neuralgia Beneficio de la toxina botulínica tipo A en neuralgia del trigemino

    Directory of Open Access Journals (Sweden)

    Carlos Zúñiga

    2008-09-01

    Full Text Available Botulinum toxin has been thoroughly studied as a potential tool in the treatment of several pain syndromes. Therefore, we assessed the clinical effects of botulinum toxin type A injections in 12 patients with otherwise unresponsive idiopathic trigeminal neuralgia. Patients were infiltrated with 20-50 units of botulinum toxin in trigger zones. Those who presented with mandibular involvement were also infiltrated in the masseter muscle. The patients were assessed on a weekly basis using the Visual Analogic Scale for pain. Ten of our patients reported a significant benefit from botulinum toxin injections, with reduction or even disappearance of pain, and remained pain free for as long as 60 days. Our findings suggest that botulinum toxin may represent a useful therapeutic tool in the management of patients with this entity.La toxina botulínica ha sido estudiada en forma exhaustiva como una potencial herramienta en el tratamiento de múltiples síndromes dolorosos. Por lo tanto, evaluamos los efectos clínicos de la aplicación de toxina botulínica tipo A en 12 sujetos con neuralgia trigeminal idiopática resistente a manejo farmacológico. Se aplicaron en dichos sujetos entre 20 y 50 unidades de toxina botulínica tipo A en las zonas gatillo. Además se infiltró el músculo masetero en aquellos que presentaban involucro mandibular. Los sujetos fueron evaluados semanalmente con una escala visual análoga para dolor. Diez de los sujetos reportaron un beneficio significativo con el uso de toxina botulínica, con reducción e incluso desaparición del dolor, permaneciendo libres de dolor por un periodo de hasta 60 días. Nuestros hallazgos sugieren que la toxina botulínica puede representar una herramienta terapéutica útil en el manejo de pacientes con esta entidad.

  14. Botulinum toxin: application, safety, and limitations.

    Science.gov (United States)

    Bigalke, Hans

    2013-01-01

    Botulinum neurotoxin type A (BoNT/A), despite its high toxicity, is approved for therapy of many neurological (e.g., dystonia, spasticity) and non-neurological (e.g., achalasia, hyperhidrosis) disorders. Its mode of action is well understood. This has led to more and more indications (e.g., pain, gastrointestinal and urologic disorders), in which the toxin can reduce disturbing symptoms. In general the application is safe (pharmacological index 20-100, depending on indication). Few unwanted reactions may occur. In worst cases BoNT treated patients may develop neutralizing antibodies. These patients are excluded from further treatment. A more recently approved second serotype (BoNT/B) could be effective in those secondary non-responders, however, due to less potency in humans higher doses have to be applied leading to an only transient successful treatment. Other serotypes as BoNT/A and B, e.g., BoNT/C should be approved as medicines. PMID:23239359

  15. Type C botulism in swine fed on restaurant waste

    Directory of Open Access Journals (Sweden)

    Djeison L. Raymundo

    2012-11-01

    Full Text Available The paper addresses the epidemiologic data of the death of pigs during the period of 2002 to 2009 following the ingestion of botulinum neurotoxin type C. This neurotoxin was present in food residues originating from restaurant and hotel kitchens, stored in barrels without shelter from the sun and administered in a collective trough without prior thermal treatment. Animals which died at different ages showed clinical signs of botulism characterized by flaccid paralysis, weight loss, anorexia, weakness, lack of coordination, locomotion difficulties with the evolution of lateral recumbency with involuntary urination and defecation. No alterations were observed at postmortem and histological examination. The bioassay with serum neutralization in mice was carried out on samples of intestinal contents from pigs affected and revealed the presence of large quantities of botulinum toxin type C.

  16. Possible origin of the high incidence of Clostridium botulinum type E in an inland bay (Green Bay of Lake Michigan).

    Science.gov (United States)

    Bott, T L; Johnson, J; Foster, E M; Sugiyama, H

    1968-05-01

    Bottom and shoreline sediments of Green Bay, northern Lake Michigan, and rivers of the Green Bay drainage basin, as well as soils of the surrounding land mass, were examined for Clostridium botulinum type E. Detection was based on identification of type E toxin in enrichment cultures and was influenced by many factors. Testing smaller amounts of sample in multiple cultures was more productive than examining large inocula in fewer cultures. Incubation at 30 C was unsatisfactory, but 14 days at 20 C or 7 days at 25 C gave good results. Mild heating (60 C for 30 min) of specimens reduced the incidence of positive findings. Freezing enrichment cultures prior to testing for toxicity eliminated many nonbotulinal toxic substances that killed mice. A control culture inoculated with type E spores was employed to show whether a specimen contained factors which could mask the presence of type E. Samples from 708 stations were tested in 2,446 cultures. Type E was found in nearly all underwater specimens of Green Bay and northern Lake Michigan but was present less frequently in samples taken along their shores. The incidence was still lower in the rivers emptying into Green Bay with the organism being rare on the shores of these rivers and in the soils of the land mass proper. Samples from the upper reaches of the rivers practically never contained type E. Runoff could deposit type E spores in Green Bay, but this is not considered to be the major factor in the high incidence of the organism. Multiplication in the bay itself is indicated. PMID:4870273

  17. Unique Ganglioside Recognition Strategies for Clostridial Neurotoxins

    Energy Technology Data Exchange (ETDEWEB)

    Benson, Marc A.; Fu, Zhuji; Kim, Jung-Ja P.; Baldwin, Michael R. (MCW); (UMC)

    2012-03-15

    Botulinum neurotoxins (BoNTs) and tetanus neurotoxin are the causative agents of the paralytic diseases botulism and tetanus, respectively. The potency of the clostridial neurotoxins (CNTs) relies primarily on their highly specific binding to nerve terminals and cleavage of SNARE proteins. Although individual CNTs utilize distinct proteins for entry, they share common ganglioside co-receptors. Here, we report the crystal structure of the BoNT/F receptor-binding domain in complex with the sugar moiety of ganglioside GD1a. GD1a binds in a shallow groove formed by the conserved peptide motif E ... H ... SXWY ... G, with additional stabilizing interactions provided by two arginine residues. Comparative analysis of BoNT/F with other CNTs revealed several differences in the interactions of each toxin with ganglioside. Notably, exchange of BoNT/F His-1241 with the corresponding lysine residue of BoNT/E resulted in increased affinity for GD1a and conferred the ability to bind ganglioside GM1a. Conversely, BoNT/E was not able to bind GM1a, demonstrating a discrete mechanism of ganglioside recognition. These findings provide a structural basis for ganglioside binding among the CNTs and show that individual toxins utilize unique ganglioside recognition strategies.

  18. Safety Evaluation of Sous Vide-Processed Products with Respect to Nonproteolytic Clostridium botulinum by Use of Challenge Studies and Predictive Microbiological Models

    OpenAIRE

    Hyytiä-Trees, Eija; Skyttä, Eija; Mokkila, Mirja; Kinnunen, Arvo; Lindström, Miia; Lähteenmäki, Liisa; Ahvenainen, Raija; Korkeala, Hannu

    2000-01-01

    Sixteen different types of sous vide-processed products were evaluated for safety with respect to nonproteolytic group II Clostridium botulinum by using challenge tests with low (2.0-log-CFU/kg) and high (5.3-log-CFU/kg) inocula and two currently available predictive microbiological models, Food MicroModel (FMM) and Pathogen Modeling Program (PMP). After thermal processing, the products were stored at 4 and 8°C and examined for the presence of botulinal spores and neurotoxin on the sell-by da...

  19. Application of botulinum toxin in the department of dermatology and pain medicine%肉毒毒素在皮肤科及疼痛科领域的应用

    Institute of Scientific and Technical Information of China (English)

    张燕; 刘桂丽; 林元珠

    2015-01-01

    Botulinum toxin is a neurotoxin derived from clostridium botulinum.It has been widely used in the treatment of neuromuscular diseases.With the research and application of botulinum toxin,botulinum toxin type A has been used with a great program in the department of dermatology and pain medicine.In this article,the advances of the botulinum toxin in the above two fields are reviewed.%肉毒毒素是肉毒杆菌产生的一种强烈神经毒素,已广泛应用于神经肌肉等疾病的治疗.随着人们对肉毒毒素的研究及应用,A型肉毒毒素在皮肤科和疼痛科领域的应用也有了长足的进展.本文概述了肉毒毒素在皮肤科及疼痛科领域的研究及应用的新进展.

  20. A proposal to prevent the "Mephisto sign" side effect of botulinum toxin type A injection in chronic migraine.

    Science.gov (United States)

    Cho, Eunae S; Hwang, Jae Young; Kim, Seong Taek

    2013-11-01

    Botulinum toxin type A (BoNT-A) has been reported as an effective treatment for chronic migraine. When BoNT-A is injected on the frontalis muscle for chronic migraine, an unexpected clinical side effect called the "Mephisto sign" may occur. The aim of this article is to propose a method to eliminate or prevent the Mephisto sign side effect. A 25-year-old female patient visited the hospital and was diagnosed with chronic migraine. A total of 155 U of BoNT-A was injected into 31 sites. 2-weeks later, and the patient developed the Mephisto sign. An additional 2-U dose was administered bilaterally to the lateral-most point of the frontalis muscles, and the eyebrow morphology returned to normal within 2-3 weeks. We propose that the development of the Mephisto sign may be prevented with an additional BoNT-A injection of 2-4 U bilaterally to the lateral most point of the frontalis muscles during the primary injection process.

  1. Factors affecting growth and toxin production by Clostridium botulinum type E on irradiated (0.3 Mrad) chicken skins

    International Nuclear Information System (INIS)

    A model system (chicken skins with chicken exudate) was used to determine if Clostridium botulinum type E (Beluga) spores, stressed by low dose irradiation, would develop and produce toxin at abuse temperatures of 10 and 300C in the absence of characteristic spoilage. Unstressed spores germinated, multiplied, and produced toxin on vacuum-packed chicken skins, stored at either 30 or 100C. Cell numbers increased faster and toxin was evident sooner at 300C than at 100C. At 300C, growth occurred and toxin was produced more slowly when samples were incubated aerobically than anaerobically. When samples were incubated aerobically at 100C, no toxin was detected within a test period of 14 days. An irradiation dose of 0.3 Mrad at 50C reduced a spore population on vacuum-sealed chicken skins by about 90%. The surviving population produced toxin at 300C under either aerobic or anaerobic conditions, at 100C no toxin was detected even on skins incubated anaerobically. Under the worst conditions (300C, vacuum packed) toxin was not detected prior to characteristic spoilage caused by the natural flora surviving 0.3 Mrad

  2. Botulinum Toxin Type A for the Treatment of Neuropathic Pain in Neuro-Rehabilitation

    Directory of Open Access Journals (Sweden)

    Domenico Intiso

    2015-06-01

    Full Text Available Pain is a natural protective mechanism and has a warning function signaling imminent or actual tissue damage. Neuropathic pain (NP results from a dysfunction and derangement in the transmission and signal processing along the nervous system and it is a recognized disease in itself. The prevalence of NP is estimated to be between 6.9% and 10% in the general population. This condition can complicate the recovery from stroke, multiple sclerosis, spinal cord lesions, and several neuropathies promoting persistent disability and poor quality of life. Subjects suffering from NP describe it as burning, itching, lancing, and numbness, but hyperalgesia and allodynia represent the most bothersome symptoms. The management of NP is a clinical challenge and several non-pharmacological and pharmacological interventions have been proposed with variable benefits. Botulinum toxin (BTX as an adjunct to other interventions can be a useful therapeutic tool for the treatment of disabled people. Although BTX-A is predominantly used to reduce spasticity in a neuro-rehabilitation setting, it has been used in several painful conditions including disorders characterized by NP. The underlying pharmacological mechanisms that operate in reducing pain are still unclear and include blocking nociceptor transduction, the reduction of neurogenic inflammation by inhibiting neural substances and neurotransmitters, and the prevention of peripheral and central sensitization. Some neurological disorders requiring rehabilitative intervention can show neuropathic pain resistant to common analgesic treatment. This paper addresses the effect of BTX-A in treating NP that complicates frequent disorders of the central and peripheral nervous system such as spinal cord injury, post-stroke shoulder pain, and painful diabetic neuropathy, which are commonly managed in a rehabilitation setting. Furthermore, BTX-A has an effect in relief pain that may characterize less common neurological disorders

  3. Toxin production of non-proteolytic Cl. botulinum type B in radurized fish. Part of a coordinated programme on the wholesomeness of the process of food irradiation

    International Nuclear Information System (INIS)

    Toxin formation by proteolytic and nonproteolytic strains of C. botulinum type B in radurized raw fish and in radurized Pindang fish was investigated. In radurized Pindang fish samples, inoculation was done either before or after cooking. Radurization process with 2 and 3 kGy caused the extension of storage life of Rastrelliger sp., Euthynnus sp., and Scomberomorus sp. by factors of 2 and 2.5 at storage temperatures between 5 and 100 C. In general at 10.5 +- 0.30 C, no toxin was formed before the samples were spoiled both in irradiated and unirradiated samples. At 5.6 +- 0.50 C no toxin was formed until after the samples were spoiled. The earliest toxin formation in unirradiated Pindang samples stored at ambient temperature was detected after the samples were spoiled. In irradiated Pindang samples inoculated with C. botulinum spores after cooking and stored at ambient temperature the toxin formation was detected before the samples were spoiled. However, if the inoculation was done before the fish was processed into Pindang, the toxin was always detected after the samples were spoiled regardless of the irradiation dose, strain and inoculation level. As fish may be contaminated - if at all - with spores of C. botulinum in its raw state, processing of fish into Pindang and irradiation would not contribute to the health hazard concerning botulism even if the samples are stored at ambient temperature

  4. Botulinum toxin injection - larynx

    Science.gov (United States)

    Injection laryngoplasty; Botox-larynx: spasmodic dysphonia-BTX; Essential voice tremor (EVT)-btx; Glottic insufficiency; Percutaneous electromyography-guided botulinum toxin treatment; Percutaneous indirect laryngoscopy-guided botulinum toxin Treatment; ...

  5. Botulinum toxin type A for cephalic cutaneous allodynia in chronic migraine: a randomized, double-blinded, placebo-controlled trial

    Directory of Open Access Journals (Sweden)

    Luciano Hollanda

    2014-12-01

    Full Text Available Cephalic allodynia (CA can be observed in 50-70% of patients with chronic migraine (CM. The aim of this trial was to assess the efficacy of botulinum toxin type A (Botx-A in the treatment of CA associated with CM. In this placebo-controlled trial, patients were randomized either into Botx-A or 0.9% saline injections and efficacy measures were assessed every 4 weeks for 3 months. Efficacy endpoints were number of migraine episodes associated with CA, changes from baseline in visual analogical scale scores for pain (VAS and frequency of common analgesics use for migraine. A total of 38 subjects were randomized to saline (n=18 or Botx-A (n=20. There were no significant differences in baseline between active intervention or placebo groups regarding mean age, number of headache episodes [mean 12.1 (9.22 and 17.00 (9.69 respectively; P=0.12], pain severity as measured by the VAS or frequency of analgesic use for headache episodes. Efficacy analysis showed that Botx-A injections led to an important decrease from baseline in the mean migraine episodes associated with CA after 12 weeks (5.20 versus 11.17; P=0.01. Also, VAS scores and frequency of analgesics use for headache were significantly reduced in the Botx-A group. This study suggests that Botx-A injections are superior to saline in the treatment of CA associated with CM, with mild self limited side effects.

  6. Multilevel botulinum toxin type a as a treatment for spasticity in children with cerebral palsy: a retrospective study

    Directory of Open Access Journals (Sweden)

    Ece Unlu

    2010-01-01

    Full Text Available INTRODUCTION: Cerebral palsy is the most common cause of physical disability in children. Spasticity is a disabling clinical symptom that is prevalent among patients suffering from cerebral palsy. The treatment of spasticity with botulinum toxin type A (BTX-A is a well-established option in the interdisciplinary management of spasticity, providing focal reductions in muscle tone in cerebral palsy patients. OBJECTIVE: The aim of this retrospective study was to describe the effect of multilevel BTX-A injections in the lower extremities, focusing mainly on gross motor function and functional status in cerebral palsy patients. METHODS: Data from 71 cerebral palsy patients (64% male, 36% female, mean age 6.7 ±3.2 years were analyzed retrospectively. We used the Ashworth and Tardieu scales to evaluate the degree of spasticity. Motor function was measured by the Gross Motor Function Measure (GMFM-88, and functional status was classified by the Gross Motor Function Classification System (GMFCS I-V. Multilevel BTX-A injections were applied after sedation and with electrostimulation guidance. The evaluations were repeated every three months, and the patients were followed for six months. RESULTS: We found that the Ashworth and Tardieu scores decreased significantly at the three-month evaluation (p0.05. Although the improvement in spasticity was not maintained at the six-month evaluation, GMFM-88 scores increased significantly at the three- and six-month assessments. GMFSC levels showed no change in the three- and six-month assessments. CONCLUSION: We believe that a single multilevel BTX-A injection reduces spasticity and improves motor function in children with cerebral palsy.

  7. Botulinum toxin type A-a novel treatment for provoked vestibulodynia? Results from a randomized, placebo controlled, double blinded study

    DEFF Research Database (Denmark)

    Petersen, Christina Damsted; Giraldi, Annamaria; Lundvall, Lene;

    2009-01-01

    receive Botox (N = 32) or saline placebo (N = 32). Botulinum toxin A (20 I.E.) diluted in 0.5 mL saline or 0.5 mL saline was injected in the musculus bulbospongiosus at baseline. MAIN OUTCOME MEASURES: Pain was measured monthly on a visual analog scale (VAS) Likert scale. Sexual function was measured...

  8. 注射用A型肉毒毒素的生物学特性及质量分析%Biological characteristics and quality of botulinum toxin type A for injection

    Institute of Scientific and Technical Information of China (English)

    何星; 苗承辉; 李小娟; 王云天; 梁琪; 张雪平; 王荫椿

    2012-01-01

    Objective To analyze the property of active pharmaceutical ingredient (API) of botulinum toxin type A (BTXA) for injection, as well as the specific activities of the preparations manufactured in 2009~2011. Methods Various components were separated from the harvested BTXA by anion exchange chromatography. The properties of BTXA complex and its components were analyzed by hemagglutination test, HPLC, SDS-PAGE, isoelectric focusing eleetrophoresis and amino acid sequencing at N-terminus. The data on specific activity of bulks of BTXA manufactured in 2009~2011 were analyzed, based on which the stability of production procedure and the reproducibility of quality were evaluated. Results The harvested BTXA was dissociated in basic environment. The obtained neurotoxin showed no hemagglutination titer, of which the relative molecular mass was about 150 000. The API of BTXA was an intact monomer, of which the purity was more than 99. 5%. The isoelectric points of BTXA complex and neurotoxin were 4. 97 and 4. 91 respectively, of which the amino acid sequences at N-terminus were basically consistent with those reported in GenBank. The mean specific activity of bulks of BTXA manufactured in 2009~2011 was 3. 0×107 LD50/mg protein. The load of API in final product of BTXA was about 5 ng per container. Conclusion The API of BTXA was a specific complex which was dissociated in basic environment to obtain neurotoxin. The specific activities of bulk BTXA manufactured in 2009~2011 were stable, indicating good persistency of quality, which provided a basis for safety of BTXA in clinic.%目的 对注射用A型肉毒毒素(Botulinum toxin type A for injection,商品名衡力,出口商品名BTXA)制品中的活性药用成分(Active pharmaceutical ingredient,API)的性质及2009 ~ 2011年生产的注射用A型肉毒毒素原液的比活性进行分析.方法 对BTXA收获物样品进行阴离子交换层析,分离毒素复合体各组分;采用血凝试验、HPLC法、SDS

  9. Studies on the irradiation of toxins of Clostridium botulinum and Staphylococcus aureus

    Energy Technology Data Exchange (ETDEWEB)

    Rose, S.A.; Bailey, N.E.; Stringer, M.F. (Campden Food and Drink Research Association, Chipping Campden (UK)); Modi, N.K.; Tranter, H.S. (Porton International plc., London (UK)); Hambleton, P. (Centre for Applied Microbiological Research, Porton Down (UK))

    1988-10-01

    The effects of irradiation of Clostridium botulinum neurotoxin type A (BNTA) and staphylococcal enterotoxin A (SEA) in gelatin phosphate buffer and cooked mince beef slurries were investigated. Estimation of toxins by immunoassays showed that in buffer, toxins were destroyed by irradiation at 8.0 kGy; in mince slurries however, 45% of BTNA and 27-34% of SEA remained after this level of irradiation. At 23.7 kGy, over twice the dose of irradiation proposed for legal acceptance in the UK, 15% of BNTA and 16-26% of SEA still remained. Increasing concentrations of mince conferred increased protection against the effect of irradiation on both toxins. The biological activity of BNTA was more sensitive to irradiation than the immunological activity. Staphylococcal enterotoxin was more resistant to irradiation than BNTA. Irradiation should therefore only be used in conjunction with good manufacturing practices to prevent microbial proliferation and toxin production prior to irradiation. (author).

  10. Studies on the irradiation of toxins of Clostridium botulinum and Staphylococcus aureus

    International Nuclear Information System (INIS)

    The effects of irradiation of Clostridium botulinum neurotoxin type A (BNTA) and staphylococcal enterotoxin A (SEA) in gelatin phosphate buffer and cooked mince beef slurries were investigated. Estimation of toxins by immunoassays showed that in buffer, toxins were destroyed by irradiation at 8.0 kGy; in mince slurries however, 45% of BTNA and 27-34% of SEA remained after this level of irradiation. At 23.7 kGy, over twice the dose of irradiation proposed for legal acceptance in the UK, 15% of BNTA and 16-26% of SEA still remained. Increasing concentrations of mince conferred increased protection against the effect of irradiation on both toxins. The biological activity of BNTA was more sensitive to irradiation than the immunological activity. Staphylococcal enterotoxin was more resistant to irradiation than BNTA. Irradiation should therefore only be used in conjunction with good manufacturing practices to prevent microbial proliferation and toxin production prior to irradiation. (author)

  11. Botulinum Toxin Type A for Treatment of Facial Wrinkles%A型肉毒毒素用于面部除皱的护理

    Institute of Scientific and Technical Information of China (English)

    党艳茹; 卢宁; 刘玉琴; 李芊

    2014-01-01

    Objective:To explore the nursing effect of the face lift and injection of botulinum toxin A treatment,in order to improve the level and effect of facial wrinkles rhytidectomy. Method:The receiving and treatment of facial wrinkles randomness extraction in 166 cases from February,2010 to December,2013 in the hospital were analyzed retrospectively,including 150 women and 16 men. They were treated with botulinum toxin type A. The treatment effect and characteristics were analyzed. Result:After the use of botulinum toxin injection in the treatment of facial wrinkles,the effect is good,effective rate of the treatment of crow’s feet is 98%,the effective rate in treatment of forehead wrinkles is 100%,and the effective rate in treatment of wrinkles between brow is 96%. Conclusion:The facial wrinkles with botulinum toxin A injection for treatment has good effect and safety,which is under convenient operation. So it is worthy of application and popularization in facial wrinkle elimination.%目的:分析针对面部除皱采用A型肉毒毒素注射治疗的护理效果和特点,以提高面部除皱的效果。方法:从我院2010年2月至2013年12月收治的面部除皱患者中随机性抽取166例进行回顾性分析和研究,其中女性150例,男性有16例,均采用A型肉毒毒素注射治疗,分析治疗的效果和特点。结果:采用肉毒毒素注射治疗面部皱纹后,效果较好,鱼尾纹的治疗有效率为98%,额部皱纹的治疗有效率为100%,眉间皱纹的治疗有效率为96%。结论:针对面部皱纹采用注射A型肉毒毒素进行治疗,效果和安全性较好,操作方便,值得在面部皱纹消除中应用和推广。

  12. A型肉毒毒素过敏事件的调查分析%Allergic reaction of the botulinum toxin type A

    Institute of Scientific and Technical Information of China (English)

    孙志成; 田茂生; 高锷; 王历; 鲁树荣

    2016-01-01

    Objective To investigate the cases of allergic reaction caused by botulinum toxin type A and the corresponding measures for its prevention and treatment.Methods The cases of allergic reaction caused by botulinum toxin type A were surveyed from 20 domestic medical cosmetic institutes from Jan.2013 to Jun.2016.Skin test was preformed for high risk population.Clinical data including customers' age,sex,injected site and times were collected in the hospitals,and cases of allergic reaction and their clinical manifestations were analyzed.Results Of 98000 cases injected with botulinum toxin type A,allergic shock occurred in 2 cases and allergic reaction in 21 cases,including wheal,redness,itching and swelling etc.Conclusions Allergic reaction caused by botulinum toxin type A is mainly related with the composition of botulinum toxin type A,cosmetics and physical constitution.Severe complications such as allergic shock should be paid high attention.Screening of the high risk population should be emphasized to avoid the allergic events.%目的 通过调查A型肉毒毒素过敏事件发生的基本情况,探讨其发生的原因及相应的防治措施.方法 调查国内20家医疗美容机构2013年1月至2016年6月A型肉毒毒素过敏反应发生情况,对高危人群进行皮试观察过敏情况.对注射A型肉毒毒素的美容就医者年龄、性别、注射部位以及被调查医疗美容机构年注射人次、有无A型肉毒毒素过敏病例,过敏病例的临床表现等进行分析.结果 9.8万人次A型肉毒毒素注射病例中,出现休克表现为2例,出现局部皮肤过敏表现为21例,皮肤过敏主要症状为局部风团、红肿、瘙痒和水肿等.A型肉毒毒素皮肤过敏反应主要与材料成分、化妆品和体质有关.结论 A型肉毒毒素过敏反应及过敏性休克等严重不良反应要引起高度重视.加强注射前高危人群筛查,避免过敏事件发生.

  13. Studies on growth and toxin production of C. botulinum type E on cod homogenate treated with a combination of spices, sodium chloride and gamma-radiation

    Energy Technology Data Exchange (ETDEWEB)

    Siddiqui, A.K. (Atomic Energy Centre, Dacca (Bangladesh)); Ando, Y.; Karashimada, T.; Kameyama, K.

    1979-09-01

    Cod homogenates inoculated with spores of C. botulinum type E strain Erimo at 10/sup 2/ and 10/sup 4//g were treated with 1% and 2% sodium chloride, 0.25% each of mustard, garlic and turmeric and 0.3 Mrad ..gamma..-radiation either in single or combination treatments. The growth and toxin production of type E spores in the inoculated homogenates were followed at incubation temperatures of 30/sup 0/, 10/sup 0/ and 5/sup 0/C for 7, 28 and 56 days respectively. Growth and gas formation were noted in all the samples but type E toxin could not be detected. The reason for the absence of toxin in both the untreated and treated homogenates could not be ascertained. Inadequate detection method, unfavourable growth conditions in the homogenate and weak toxigenicity of the strain employed have been advanced as probable factors that contributed to the negative results on the toxin assay.

  14. Phospholipase C Produced by Clostridium botulinum Types C and D:Comparison of Gene, Enzymatic, and Biological Activities with Those of Clostridium perfringens Alpha-toxin

    Directory of Open Access Journals (Sweden)

    Sakurai,Jun

    2013-02-01

    Full Text Available Clostridium botulinum type C and D strains recently have been found to produce PLC on egg yolk agar plates. To characterize the gene, enzymatic and biological activities of C. botulinum PLCs (Cb-PLCs, the cb-plc genes from 8 strains were sequenced, and 1 representative gene was cloned and expressed as a recombinant protein. The enzymatic and hemolytic activities of the recombinant Cb-PLC were measured and compared with those of the Clostridium perfringens alpha-toxin. Each of the eight cb-plc genes encoded a 399 amino acid residue protein preceded by a 27 residue signal peptide. The protein consists of 2 domains, the N- and C-domains, and the overall amino acid sequence identity between Cb-PLC and alpha-toxin was greater than 50%, suggesting that Cb-PLC is homologous to the alpha-toxin. The key residues in the N-domain were conserved, whereas those in the C-domain which are important in membrane interaction were different than in the alpha-toxin. As expected, Cb-PLC could hydrolyze egg yolk phospholipid, p-nitrophenylphosphorylcholine, and sphingomyelin, and also exhibited hemolytic activity;however, its activities were about 4- to over 200-fold lower than those of alpha-toxin. Although Cb-PLC showed weak enzymatic and biological activities, it is speculated that Cb-PLC might play a role in the pathogenicity of botulism or for bacterial survival.

  15. Mechanisms of food processing and storage-related stress tolerance in Clostridium botulinum.

    Science.gov (United States)

    Dahlsten, Elias; Lindström, Miia; Korkeala, Hannu

    2015-05-01

    Vegetative cultures of Clostridium botulinum produce the extremely potent botulinum neurotoxin, and may jeopardize the safety of foods unless sufficient measures to prevent growth are applied. Minimal food processing relies on combinations of mild treatments, primarily to avoid deterioration of the sensory qualities of the food. Tolerance of C. botulinum to minimal food processing is well characterized. However, data on effects of successive treatments on robustness towards further processing is lacking. Developments in genetic manipulation tools and the availability of annotated genomes have allowed identification of genetic mechanisms involved in stress tolerance of C. botulinum. Most studies focused on low temperature, and the importance of various regulatory mechanisms in cold tolerance of C. botulinum has been demonstrated. Furthermore, novel roles in cold tolerance were shown for metabolic pathways under the control of these regulators. A role for secondary oxidative stress in tolerance to extreme temperatures has been proposed. Additionally, genetic mechanisms related to tolerance to heat, low pH, and high salinity have been characterized. Data on genetic stress-related mechanisms of psychrotrophic Group II C. botulinum strains are scarce; these mechanisms are of interest for food safety research and should thus be investigated. This minireview encompasses the importance of C. botulinum as a food safety hazard and its central physiological characteristics related to food-processing and storage-related stress. Special attention is given to recent findings considering genetic mechanisms C. botulinum utilizes in detecting and countering these adverse conditions.

  16. [Intoxication of botulinum toxin].

    Science.gov (United States)

    Chudzicka, Aleksandra

    2015-09-01

    Botulinum toxin is an egzotoxin produced by Gram positive bacteria Clostridium botulinum. It is among the most potent toxins known. The 3 main clinical presentations of botulism are as follows: foodborne botulism, infant botulism and wound botulism. The main symptom of intoxication is flat muscles paralysis. The treatment is supportive care and administration of antitoxin. In prevention the correct preparing of canned food is most important. Botulinum toxin is accepted as a biological weapon.

  17. Synaptobrevin/vesicle-associated membrane protein (VAMP) of Aplysia californica: structure and proteolysis by tetanus toxin and botulinal neurotoxins type D and F.

    Science.gov (United States)

    Yamasaki, S; Hu, Y; Binz, T; Kalkuhl, A; Kurazono, H; Tamura, T; Jahn, R; Kandel, E; Niemann, H

    1994-01-01

    Synaptobrevin/vesicle-associated membrane protein (VAMP) and syntaxin are potential vesicle donor and target membrane receptors of a docking complex that requires N-ethylmaleimide-sensitive factor (NSF) and soluble NSF-attachment proteins as soluble factors for vesicle fusion with target membranes. Members of this docking complex are the target of clostridial neurotoxins that act as zinc-dependent proteases. Molecular cloning of the Aplysia californica synaptobrevin cDNA revealed a 180-residue polypeptide (M(r), 19,745) with a central transmembrane region and an atypically large C-terminal intravesicular domain. This polypeptide integrates into membranes at both the co- and posttranslational level, as shown by modification of an artificially introduced N-glycosylation site. The soluble and membrane-anchored forms of synaptobrevin are cleaved by the light chains of the botulinal toxins type D and F and by tetanus toxin involving the peptide bonds Lys49-Ile50, Gln48-Lys49, and Gln66-Phe67, respectively. The active center of teh tetanus toxin light chain was identified by site-specific mutagenesis. His233, His237, Glu234, and Glu270/271 are essential to this proteolytic activity. Modification of histidine residues resulted in loss of zinc binding, whereas a replacement of Glu234 only slightly reduced the zinc content. Images PMID:8197120

  18. Use of Botulinum Neurotoxin for the Treatment of Movement Disorders

    Science.gov (United States)

    ... of muscles. Neurologists from the American Academy of Neurology are doctors who identify and treat diseases of ... evidence-based information* is provided by experts in neurology who carefully reviewed all available scientific studies on ...

  19. Use of Botulinum Neurotoxin Injections to Treat Spasticity

    Science.gov (United States)

    ... and pain. Neurologists from the American Academy of Neurology (AAN) are doctors who identify and treat diseases ... evidence-based information* is provided by experts in neurology who carefully reviewed all available evidence on the ...

  20. Technologies for detecting botulinum neurotoxins in biological and environmental matrices

    Science.gov (United States)

    Biomonitoring of food and environmental matrices is critical for the rapid and sensitive diagnosis, treatment, and prevention of diseases caused by toxins. The United States Centers for Disease Control and Prevention (CDC) has noted that toxins from bacteria, fungi, algae, and plants present an ongo...

  1. Botulinum Toxin Type A Injection Combined With Cast Immobilization for Treating Recurrent Peroneal Spastic Flatfoot Without Bone Coalitions: A Case Report and Review of the Literature.

    Science.gov (United States)

    Xu, Jian; Muhammad, Hassan; Wang, Xu; Ma, Xin

    2015-01-01

    Peroneal spastic flatfoot is an uncommon condition. It often presents as a rigid and usually painful valgus deformity in the hindfoot with peroneal muscles spasms. Although tarsal coalition is an important cause, a few patients have not undergone bone coalitions. We describe a 27-year-old female who experienced recurrent peroneal spastic flatfoot after an injury. She was treated successfully with a combination of botulinum toxin type A and immobilization of the foot in a neutral position with a cast. After 3 years, the condition had not recurred, and she was pain free and walked normally, with no increase in muscle tone. This unique treatment could be of potential use to treat many patients with such conditions.

  2. A 型肉毒毒素 ELISA 鉴别试验方法的建立%Establishment of an ELISA identification for botulinum toxin type A

    Institute of Scientific and Technical Information of China (English)

    萧在澜; 王媛; 顾磊; 杨雨生; 赵建荣; 徐永浩; 张华捷

    2015-01-01

    Objective To establish a new identification method for botulinum toxin type A ( botA) to replace the traditional animal test.Methods A double antibody sandwich ELISA was established based on the modern immunological technolo-gy, by using prepared two kinds of anti-botA polyclonal antibodies derived from equine and rabbit origin, and a preliminary verification test was performed.Results The established identification ELISA could be a potential alternative method in re-placement of animal test, it is of good specificity, sensitivity, precision and robustness.Conclusion The established method will be expected to be approved for the identification test of botulinum toxin type A as the replacement method for traditional animal test by a further verification and validation.%目的:建立鉴定A型肉毒毒素的ELISA鉴别试验方法以替代传统的动物试验法。方法采用现代免疫学技术,制备马源性和兔源性抗A型肉毒毒素多克隆抗体,建立了双抗体夹心ELISA,并就此初步进行方法学验证。结果所建立的ELISA具有良好的特异性、灵敏度、精密度和耐用性,具有替代动物试验方法的良好前景。结论在进一步验证和确认之后,该方法有望正式成为可用于鉴定A型肉毒毒素的试验方法以替代动物试验法。

  3. [Dose-response relationship in the treatment of cervical dystonia with botulinum toxin type A (AGN 191622)--a phase II study].

    Science.gov (United States)

    Mezaki, T; Kaji, R; Kimura, J; Mannen, T

    1995-09-01

    Injection of botulinum toxin type A has been the treatment of choice for spasmodic torticollis for several years. Although previous reports demonstrate its effectiveness and safety, the treatment strategy has been empirical. The present study, using the freeze-dried crystalline botulinum toxin type A (AGN 191622; Allergan Inc., Irvine, CA), aimed to compare the efficacy among three treatment groups divided into low, medium and high dosage levels. Fifty-one patients who entered the study were grouped into low-dose (60 units/session), medium-dose (120 units/session) and high-dose (240 units/session) groups. Two patients (one in low-dose group and the other in high-dose group) were excluded from the assessment of efficacy because they dropped out in the early phase of the study. One experienced worsening of an existing psychosis and the other developed an acute respiratory infection. Injection sites were decided individually by palpation. If the clinical response was not satisfactory four weeks after an injection, the patient was re-injected with the same dose of toxin. The follow-up period was 14 weeks from the initial injection. The results showed that the high-dose group improved more than the other groups in the parameters of severity of symptoms and subjective benefit (p = 0.000). Also, fewer injections were required in the high-dose group to achieve substantial clinical benefit. Although the mean reduction in Tsui's score was not statistically significant among the groups, the "marked improvement" was seen more frequently in the high-dose group (p = 0.033). Unfavorable adverse effects including excessive weakness and dysphasia were always mild and transient.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. Botulinum Toxin and Muscle Atrophy: A Wanted or Unwanted Effect.

    Science.gov (United States)

    Durand, Paul D; Couto, Rafael A; Isakov, Raymond; Yoo, Donald B; Azizzadeh, Babak; Guyuron, Bahman; Zins, James E

    2016-04-01

    While the facial rejuvenating effect of botulinum toxin type A is well known and widespread, its use in body and facial contouring is less common. We first describe its use for deliberate muscle volume reduction, and then document instances of unanticipated and undesirable muscle atrophy. Finally, we investigate the potential long-term adverse effects of botulinum toxin-induced muscle atrophy. Although the use of botulinum toxin type A in the cosmetic patient has been extensively studied, there are several questions yet to be addressed. Does prolonged botulinum toxin treatment increase its duration of action? What is the mechanism of muscle atrophy and what is the cause of its reversibility once treatment has stopped? We proceed to examine how prolonged chemodenervation with botulinum toxin can increase its duration of effect and potentially contribute to muscle atrophy. Instances of inadvertent botulinum toxin-induced atrophy are also described. These include the "hourglass deformity" secondary to botulinum toxin type A treatment for migraine headaches, and a patient with atrophy of multiple facial muscles from injections for hemifacial spasm. Numerous reports demonstrate that muscle atrophy after botulinum toxin type A treatment occurs and is both reversible and temporary, with current literature supporting the notion that repeated chemodenervation with botulinum toxin likely responsible for both therapeutic and incidental temporary muscle atrophy. Furthermore, duration of response may be increased with subsequent treatments, thus minimizing frequency of reinjection. Practitioners should be aware of the temporary and reversible effect of botulinum toxin-induced muscle atrophy and be prepared to reassure patients on this matter. PMID:26780946

  5. Tratamiento del Síndrome de Dolor Miofascial con Toxina Botulínica tipo A Botulinum toxin type A in the management of Myofascial pain syndrome

    Directory of Open Access Journals (Sweden)

    M. Castro

    2006-03-01

    during 48 hours following the injection that improved spontaneously. The satisfaction was excellent in 10 patients (50%, good in 7 (35% and average in 3 (15%. None of the patient qualified the experience as bad. Conclusion: The muscular injection of botulinum toxin type A in the treatment of MSP is effective and safe.

  6. Interaction of Botulinum Toxin with the Epithelial Barrier

    Directory of Open Access Journals (Sweden)

    Yukako Fujinaga

    2010-01-01

    Full Text Available Botulinum neurotoxin (BoNT is a protein toxin (~150 kDa, which possesses a metalloprotease activity. Food-borne botulism is manifested when BoNT is absorbed from the digestive tract to the blood stream and enters the peripheral nerves, where the toxin cleaves core proteins of the neuroexocytosis apparatus and elicits the inhibition of neurotransmitter release. The initial obstacle to orally ingested BoNT entering the body is the epithelial barrier of the digestive tract. Recent cell biology and molecular biology studies are beginning to elucidate the mechanism by which this large protein toxin crosses the epithelial barrier. In this review, we provide an overview of the structural features of botulinum toxins (BoNT and BoNT complex and the interaction of these toxins with the epithelial barrier.

  7. A型肉毒素治疗面部皱纹的临床疗效观察%The Observation effect of Botulinum Toxin type A injections in treatment of facial wrinkle

    Institute of Scientific and Technical Information of China (English)

    袁亮飞; 刘静; 刘安久; 曹进军

    2012-01-01

      Objective:To observe the effect of Botulinum Toxin type A injections in treatment of facial wrinkle. Methods:A total of 91 patients were included. Botulinum Toxin type A inject their facial wrinkle,the different local with different doses,fixed uniform inject. Results:91 patients were satisfied with the treatment. Conclusions:Botulinum Toxin type A injections therapy is a effective methods for treatment facial wrinkle. The therapeutic effect is obvious ,rapid and the side effects minimal.%  目的观察A型肉毒素(BTXA)治疗面部皱纹的疗效。方法对91例面部皱纹患者利用A型肉毒素根据部位不同,分别采用不同剂量,定点均匀注射。结果91例患者经治疗后均获得满意疗效。结论 A型肉毒素在面部除皱中有确切的疗效,且痛苦小、见效快、副作用少。

  8. Possible Origin of the High Incidence of Clostridium botulinum Type E in an Inland Bay (Green Bay of Lake Michigan)1

    Science.gov (United States)

    Bott, Thomas L.; Johnson, Jodie; Foster, E. M.; Sugiyama, H.

    1968-01-01

    Bottom and shoreline sediments of Green Bay, northern Lake Michigan, and rivers of the Green Bay drainage basin, as well as soils of the surrounding land mass, were examined for Clostridium botulinum type E. Detection was based on identification of type E toxin in enrichment cultures and was influenced by many factors. Testing smaller amounts of sample in multiple cultures was more productive than examining large inocula in fewer cultures. Incubation at 30 C was unsatisfactory, but 14 days at 20 C or 7 days at 25 C gave good results. Mild heating (60 C for 30 min) of specimens reduced the incidence of positive findings. Freezing enrichment cultures prior to testing for toxicity eliminated many nonbotulinal toxic substances that killed mice. A control culture inoculated with type E spores was employed to show whether a specimen contained factors which could mask the presence of type E. Samples from 708 stations were tested in 2,446 cultures. Type E was found in nearly all underwater specimens of Green Bay and northern Lake Michigan but was present less frequently in samples taken along their shores. The incidence was still lower in the rivers emptying into Green Bay with the organism being rare on the shores of these rivers and in the soils of the land mass proper. Samples from the upper reaches of the rivers practically never contained type E. Runoff could deposit type E spores in Green Bay, but this is not considered to be the major factor in the high incidence of the organism. Multiplication in the bay itself is indicated. PMID:4870273

  9. Substrate recognition mechanism of VAMP/synaptobrevin-cleaving clostridial neurotoxins.

    Science.gov (United States)

    Sikorra, Stefan; Henke, Tina; Galli, Thierry; Binz, Thomas

    2008-07-25

    Botulinum neurotoxins (BoNTs) and tetanus neurotoxin (TeNT) inhibit neurotransmitter release by proteolyzing a single peptide bond in one of the three soluble N-ethylmaleimide-sensitive factor attachment protein receptors SNAP-25, syntaxin, and vesicle-associated membrane protein (VAMP)/synaptobrevin. TeNT and BoNT/B, D, F, and G of the seven known BoNTs cleave the synaptic vesicle protein VAMP/synaptobrevin. Except for BoNT/B and TeNT, they cleave unique peptide bonds, and prior work suggested that different substrate segments are required for the interaction of each toxin. Although the mode of SNAP-25 cleavage by BoNT/A and E has recently been studied in detail, the mechanism of VAMP/synaptobrevin proteolysis is fragmentary. Here, we report the determination of all substrate residues that are involved in the interaction with BoNT/B, D, and F and TeNT by means of systematic mutagenesis of VAMP/synaptobrevin. For each of the toxins, three or more residues clustered at an N-terminal site remote from the respective scissile bond are identified that affect solely substrate binding. These exosites exhibit different sizes and distances to the scissile peptide bonds for each neurotoxin. Substrate segments C-terminal of the cleavage site (P4-P4') do not play a role in the catalytic process. Mutation of residues in the proximity of the scissile bond exclusively affects the turnover number; however, the importance of individual positions at the cleavage sites varied for each toxin. The data show that, similar to the SNAP-25 proteolyzing BoNT/A and E, VAMP/synaptobrevin-specific clostridial neurotoxins also initiate substrate interaction, employing an exosite located N-terminal of the scissile peptide bond.

  10. Thermal and Pressure-Assisted Thermal Destruction Kinetics for Spores of Type A Clostridium botulinum and Clostridium sporogenes PA3679.

    Science.gov (United States)

    Reddy, N Rukma; Patazca, Eduardo; Morrissey, Travis R; Skinner, Guy E; Loeza, Viviana; Schill, Kristin M; Larkin, John W

    2016-02-01

    The purpose of this study was to determine the inactivation kinetics of the spores of the most resistant proteolytic Clostridium botulinum strains (Giorgio-A and 69-A, as determined from an earlier screening study) and of Clostridium sporogenes PA3679 and to compare the thermal and pressure-assisted thermal resistance of these spores. Spores of these strains were prepared using a biphasic medium method. C. sporogenes PA3679 spores were heat treated before spore preparation. Using laboratory-scale and pilot-scale pressure test systems, spores of Giorgio-A, 69-A, and PA3679 suspended in ACES [N-(2-acetamido)-2-aminoethanesulfonic acid] buffer (pH 7.0) were exposed to various combinations of temperature (93 to 121°C) and pressure (0.1 to 750 MPa) to determine their resistance. More than a 5-log reduction occurred after 3 min at 113°C for spores of Giorgio-A and 69-A and after 5 min at 117°C for spores of PA3679. A combination of high temperatures (93 to 121°C) and pressures yielded greater log reductions of spores of Giorgio-A, 69-A, and PA3679 compared with reduction obtained with high temperatures alone. No survivors from initial levels (>5.0 log CFU) of Giorgio-A and 69-A were detected when processed at a combination of high temperature (117 and 121°C) and high pressure (600 and 750 MPa) for 4.5-log reduction of PA3679 spores. Thermal D-values of Giorgio-A, 69-A, and PA3679 spores decreased (i.e., 29.1 to 0.33 min for Giorgio-A, 40.5 to 0.27 min for 69-A, and 335.2 to 2.16 min for PA3679) as the temperature increased from 97 to 117°C. Pressure-assisted thermal D-values of Giorgio-A, 69-A, and PA3679 also decreased as temperature increased from 97 to 121°C at both pressures (600 and 750 MPa) (i.e., 17.19 to 0.15 min for Giorgio-A, 9.58 to 0.15 min for 69-A, and 12.93 to 0.33 min for PA3679 at 600 MPa). At higher temperatures (117 or 121°C), increasing pressure from 600 to 750 MPa had an effect on pressure-assisted thermal D-values of PA3679 (i.e., at 117

  11. Practical guidance for CD management involving treatment of botulinum toxin: a consensus statement

    OpenAIRE

    Albanese, Alberto; Abbruzzese, Giovanni; Dressler, Dirk; Duzynski, Wojciech; Khatkova, Svetlana; Marti, Maria-José; Mir, Pablo; Montecucco, Cesare; Moro, Elena; Pinter, Michaela; Relja, Maja; Roze, Emmanuel; Skogseid, Inger Marie; Timerbaeva, Sofiya; Tzoulis, Charalampos

    2015-01-01

    Cervical dystonia is a neurological movement disorder causing abnormal posture of the head. It may be accompanied by involuntary movements which are sometimes tremulous. The condition has marked effects on patients’ self-image, and adversely affects quality of life, social relationships and employment. Botulinum neurotoxin (BoNT) is the treatment of choice for CD and its efficacy and safety have been extensively studied in clinical trials. However, current guidelines do not provide enough pra...

  12. Striving for more good days: patient perspectives on botulinum toxin for the treatment of cervical dystonia

    OpenAIRE

    Taylor, Joanna; Poliziani,Michele; Liu, Xierong; Koch, Marco

    2016-01-01

    Michele Poliziani,1 Marco Koch,2 Xierong Liu1 1Opinion Health, London, UK; 2Merz Pharmaceuticals GmbH, Frankfurt am Main, Germany Background: The recommended reinjection interval for botulinum neurotoxin (BoNT) formulations in the treatment of cervical dystonia (CD) is generally ≥12 weeks, though intervals ≥10 weeks are approved for incobotulinumtoxinA in Europe. However, recurring symptoms can occur before the end of this period. Using qualitative research, we sought a greate...

  13. Identification and genetic characterization of Clostridium botulinum serotype A strains from commercially pasteurized carrot juice.

    Science.gov (United States)

    Marshall, Kristin M; Nowaczyk, Louis; Raphael, Brian H; Skinner, Guy E; Rukma Reddy, N

    2014-12-01

    Clostridium botulinum is an important foodborne pathogen capable of forming heat resistant endospores and producing deadly botulinum neurotoxins (BoNTs). In 2006, C. botulinum was responsible for an international outbreak of botulism attributed to the consumption of commercially pasteurized carrot juice. The purpose of this study was to isolate and characterize strains of C. botulinum from the adulterated product. Carrot juice bottles retrieved from the manufacturing facility were analyzed for the presence of BoNT and BoNT-producing isolates using DIG-ELISA. Toxigenic isolates from the carrot juice were analyzed using pulsed-field gel electrophoresis (PFGE) and DNA microarray analysis to determine their genetic relatedness to the original outbreak strains CDC51348 and CDC51303. PFGE revealed that isolates CJ4-1 and CJ10-1 shared an identical pulsotype with strain CDC51303, whereas isolate CJ5-1 displayed a unique restriction banding pattern. DNA microarray analysis identified several phage related genes unique to strain CJ5-1, and Southern hybridization analysis of XhoI digested and nondigested DNA showed their chromosomal location, while a homolog to pCLI_A009 of plasmid pCLI of C. botulinum serotype Langeland F, was located on a small plasmid. The acquisition or loss of bacteriophages and other mobile genetic elements among C. botulinum strains has epidemiological and evolutionary implications.

  14. Systematic Assessment of Nonproteolytic Clostridium botulinum Spores for Heat Resistance

    Science.gov (United States)

    Stringer, Sandra C.; Barker, Gary C.; Peck, Michael W.

    2016-01-01

    ABSTRACT Heat treatment is an important controlling factor that, in combination with other hurdles (e.g., pH, aw), is used to reduce numbers and prevent the growth of and associated neurotoxin formation by nonproteolytic C. botulinum in chilled foods. It is generally agreed that a heating process that reduces the spore concentration by a factor of 106 is an acceptable barrier in relation to this hazard. The purposes of the present study were to review the available data relating to heat resistance properties of nonproteolytic C. botulinum spores and to obtain an appropriate representation of parameter values suitable for use in quantitative microbial risk assessment. In total, 753 D values and 436 z values were extracted from the literature and reveal significant differences in spore heat resistance properties, particularly those corresponding to recovery in the presence or absence of lysozyme. A total of 503 D and 338 z values collected for heating temperatures at or below 83°C were used to obtain a probability distribution representing variability in spore heat resistance for strains recovered in media that did not contain lysozyme. IMPORTANCE In total, 753 D values and 436 z values extracted from literature sources reveal significant differences in spore heat resistance properties. On the basis of collected data, two z values have been identified, z = 7°C and z = 9°C, for spores recovered without and with lysozyme, respectively. The findings support the use of heat treatment at 90°C for 10 min to reduce the spore concentration by a factor of 106, providing that lysozyme is not present during recovery. This study indicates that greater heat treatment is required for food products containing lysozyme, and this might require consideration of alternative recommendation/guidance. In addition, the data set has been used to test hypotheses regarding the dependence of spore heat resistance on the toxin type and strain, on the heating technique used, and on the

  15. Tratamento da doença de Hailey-Hailey com toxina botulínica tipo A Hailey-Hailey disease treatment with Botulinum toxin type A

    Directory of Open Access Journals (Sweden)

    Giancarlo Rezende Bessa

    2010-10-01

    Full Text Available Duas irmãs com doença de Hailey-Hailey, com lesões recorrentes - uma em axilas e outra em região inguinal -, e resposta limitada aos tratamentos clássicos. Elas foram tratadas com aplicação de toxina botulínica tipo A. Observamos que houve importante melhora na paciente tratada na região inguinal e remissão completa na paciente em cujas axilas sofreram tratamento. Além disso, foi possível poupar uso de antibióticos sistêmicos e corticoides tópicos. O alto custo é um fator restritivo para uso rotineiro e estudos maiores são necessários para definir eficácia e relação custo-benefício dessa intervenção.Two sisters with recurrent lesions, one on axillae and other on the groin, and with limited response to classical treatments were treated with injections botulinum toxin type A. We observed marked improvement in the patient treated in the groin and complete remission in the patient treated in the axillae. It was possible to spare the use of systemic antibiotics and topical corticosteroids. The high cost is a restrictive factor to routine use and large studies are necessary to access efficacy and cost benefit profile.

  16. Ultrasound-Guided Injection of Botulinum Toxin Type A for Piriformis Muscle Syndrome: A Case Report and Review of the Literature

    Directory of Open Access Journals (Sweden)

    Andrea Santamato

    2015-08-01

    Full Text Available Piriformis muscle syndrome (PMS is caused by prolonged or excessive contraction of the piriformis muscle associated with pain in the buttocks, hips, and lower limbs because of the close proximity to the sciatic nerve. Botulinum toxin type A (BoNT-A reduces muscle hypertonia as well as muscle contracture and pain inhibiting substance P release and other inflammatory factors. BoNT-A injection technique is important considering the difficult access of the needle for deep location, the small size of the muscle, and the proximity to neurovascular structures. Ultrasound guidance is easy to use and painless and several studies describe its use during BoNT-A administration in PMS. In the present review article, we briefly updated current knowledge regarding the BoNT therapy of PMS, describing also a case report in which this syndrome was treated with an ultrasound-guided injection of incobotulinumtoxin A. Pain reduction with an increase of hip articular range of motion in this patient with PMS confirmed the effectiveness of BoNT-A injection for the management of this syndrome.

  17. The Efficacy and Safety of Fractional CO2 Laser Combined with Topical Type A Botulinum Toxin for Facial Rejuvenation: A Randomized Controlled Split-Face Study

    Directory of Open Access Journals (Sweden)

    Jie Zhu

    2016-01-01

    Full Text Available Objective. We evaluated synergistic efficacy and safety of combined topical application of Botulinum Toxin Type A (BTX-A with fractional CO2 laser for facial rejuvenation. Methods. Twenty female subjects were included for this split-face comparative study. One side of each subject’s cheek was treated with fractional CO2 plus saline solution, and the other side was treated with fractional CO2 laser plus topical application of BTX-A. Patients received one session of treatment and evaluations were done at baseline, one, four, and twelve weeks after treatment. The outcome assessments included subjective satisfaction scale; blinded clinical assessment; and the biophysical parameters of roughness, elasticity, skin hydration, transepidermal water loss (TEWL, and the erythema and melanin index. Results. BTX-A combined with fractional CO2 laser sides showed higher physician’s global assessment score, subject satisfaction score, roughness, skin hydration, and skin elasticity compared to that of fractional CO2 plus saline solution side at 12 weeks after treatment. TEWL and erythema and melanin index showed no significant differences between two sides at baseline, one, four, and twelve weeks after treatment. Conclusion. Topical application of BTX-A could enhance the rejuvenation effect of fractional CO2 laser.

  18. Study design and methods of the BoTULS trial: a randomised controlled trial to evaluate the clinical effect and cost effectiveness of treating upper limb spasticity due to stroke with botulinum toxin type A

    Directory of Open Access Journals (Sweden)

    Graham Laura

    2008-10-01

    Full Text Available Abstract Background Following a stroke, 55–75% of patients experience upper limb problems in the longer term. Upper limb spasticity may cause pain, deformity and reduced function, affecting mood and independence. Botulinum toxin is used increasingly to treat focal spasticity, but its impact on upper limb function after stroke is unclear. The aim of this study is to evaluate the clinical and cost effectiveness of botulinum toxin type A plus an upper limb therapy programme in the treatment of post stroke upper limb spasticity. Methods Trial design : A multi-centre open label parallel group randomised controlled trial and economic evaluation. Participants : Adults with upper limb spasticity at the shoulder, elbow, wrist or hand and reduced upper limb function due to stroke more than 1 month previously. Interventions : Botulinum toxin type A plus upper limb therapy (intervention group or upper limb therapy alone (control group. Outcomes : Outcome assessments are undertaken at 1, 3 and 12 months. The primary outcome is upper limb function one month after study entry measured by the Action Research Arm Test (ARAT. Secondary outcomes include: spasticity (Modified Ashworth Scale; grip strength; dexterity (Nine Hole Peg Test; disability (Barthel Activities of Daily Living Index; quality of life (Stroke Impact Scale, Euroqol EQ-5D and attainment of patient-selected goals (Canadian Occupational Performance Measure. Health and social services resource use, adverse events, use of other antispasticity treatments and patient views on the treatment will be compared. Participants are clinically reassessed at 3, 6 and 9 months to determine the need for repeat botulinum toxin type A and/or therapy. Randomisation : A web based central independent randomisation service. Blinding : Outcome assessments are undertaken by an assessor who is blinded to the randomisation group. Sample size : 332 participants provide 80% power to detect a 15% difference in treatment

  19. Botulinum toxin type A in refractory chronic migraine: an open-label trial Toxina botulínica tipo A no tratamento da enxaqueca refratária: um estudo aberto

    OpenAIRE

    Carla Menezes; Bernardo Rodrigues; Elza Magalhães; Ailton Melo

    2007-01-01

    Botulinum toxin type A (BT-A) has been described as an important strategy to various types of pain such as cervical dystonia, myofascial pain syndrome and headache. Although BT-A efficacy has not been proven in tension type headache, its use in migraine continues controversial. In this open trial, we evaluated the efficacy of BT-A in refractory migraine. BT-A was injected in patients diagnosed with migraine who had previously used three classes of prophylactic drugs by at least one year with ...

  20. Functional characterisation of germinant receptors in Clostridium botulinum and Clostridium sporogenes presents novel insights into spore germination systems.

    Science.gov (United States)

    Brunt, Jason; Plowman, June; Gaskin, Duncan J H; Itchner, Manoa; Carter, Andrew T; Peck, Michael W

    2014-09-01

    Clostridium botulinum is a dangerous pathogen that forms the highly potent botulinum toxin, which when ingested causes a deadly neuroparalytic disease. The closely related Clostridium sporogenes is occasionally pathogenic, frequently associated with food spoilage and regarded as the non-toxigenic equivalent of Group I C. botulinum. Both species form highly resistant spores that are ubiquitous in the environment and which, under favourable growth conditions germinate to produce vegetative cells. To improve the control of botulinum neurotoxin-forming clostridia, it is imperative to comprehend the mechanisms by which spores germinate. Germination is initiated following the recognition of small molecules (germinants) by a specific germinant receptor (GR) located in the spore inner membrane. The present study precisely defines clostridial GRs, germinants and co-germinants. Group I C. botulinum ATCC3502 contains two tricistronic and one pentacistronic GR operons, while C. sporogenes ATCC15579 has three tricistronic and one tetracistronic GR operons. Insertional knockout mutants, allied with characterisation of recombinant GRs shows for the first time that amino acid stimulated germination in C. botulinum requires two tri-cistronic encoded GRs which act in synergy and cannot function individually. Spore germination in C. sporogenes requires one tri-cistronic GR. Two other GRs form part of a complex involved in controlling the rate of amino-acid stimulated germination. The suitability of using C. sporogenes as a substitute for C. botulinum in germination studies and food challenge tests is discussed. PMID:25210747

  1. Systemic colonization of clover (Trifolium repens by Clostridium botulinum strain 2301

    Directory of Open Access Journals (Sweden)

    Matthias eZeiller

    2015-10-01

    Full Text Available In recent years, cases of botulism in cattle and other farm animals and also in farmers increased dramatically. It was proposed, that these cases could be affiliated with the spreading of compost or other organic manures contaminated with Clostridium botulinum spores on farm land. Thus, soils and fodder plants and finally farm animals could be contaminated. Therefore, the colonization behavior and interaction of the botulinum neurotoxin (BoNT D producing C. botulinum strain 2301 and the non-toxin producing Clostridium sporogenes strain 1739 were investigated on clover (Trifolium repens in a field experiment as well as in phytochamber experiments applying axenic and additionally soil based systems under controlled conditions. Plants were harvested and divided into root and shoot parts for further DNA isolation and PCR assays; subsamples were fixed for fluorescence in situ hybridization (FISH analysis in combination with confocal laser scanning microscopy (CLSM. To target C. botulinum and C. sporogenes, 16S rDNA directed primers were used and to specifically detect C. botulinum, BoNT D toxin genes targeted primers, using a multiplex PCR approach, were applied. Our results demonstrate an effective colonization of roots and shoots of clover by C. botulinum strain 2301 and C. sporogenes strain 1739. Detailed analysis of colonization behavior showed that C. botulinum can occur as individual cells, in cell clusters and in microcolonies within the rhizosphere, lateral roots and within the roots tissue of clover. In addition, we observed significant differences in the growth behavior of clover plants when inoculated with Clostridia spores, indicating a plant growth promoting effect. Inoculated plants showed an increased growth index (shoot size, wet and dry weight and an enlarged root system, which suggests the involvement of phytohormonal effects induced by the systemic colonization of clover by C. botulinum strain 2301.

  2. The Efficacy of Botulinum Toxin Type a Injection in the Hamstring and Calf Muscles With and Without Serial Foot Casting in Gait Improvement in Children With Cerebral Palsy

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    Shamsodini A

    2011-11-01

    Full Text Available Background: The goal of this study was to compare the efficacy of botulinum toxin type A (BTX-A injection in the hamstring and calf muscles with and without ankle serial casting in the improvement of gait in children with cerebral palsy (CP.Methods : This double-blind prospective clinical trial was performed on 25, 2 to 8-year-old children with hemiplegic or diplegic CP in Tehran, Iran in 2010. The participants were chosen by simple randomized sampling and were matched for age, gross motor function classification system (GMFCS and type of CP and were randomly divided into two groups: children in the first group (13 only received BTX-A injection, but the second group (12 received BTX-A and serial foot casting starting one week after the injection.Results : Comparison of the gross motor function, right and left knee spasticities and passive ROM of both knees between the two groups before and 1, 3, 6 and 12 months after the injections were not statistically significant (P>0.1. Furthermore, comparison of the right and left ankle spasticities and passive ROM before the injections and in1 and 3-month follow-ups did not show a statistically significant difference (P>0.1, but the differences were significant in 6 and 12-month follow-ups (P<0.05.Conclusion: BTX-A injection with serial foot casting vs. BTX-A alone was more effective in decreasing spasticity and improving passive ROM in the ankle of children with CP, but such injections in the hamstrings were not useful in these regards.

  3. Treatment of scar contracture with intralesional botulinum toxin type A injection%A型肉毒毒素治疗挛缩性瘢痕

    Institute of Scientific and Technical Information of China (English)

    颜彤彤; 陈敏亮; 马奎; 梁黎明; 刘畅; 赖林英; 付小兵

    2013-01-01

    Objective To evaluate the validity of botulinum toxin type A (BTXA) injections for the treatment of scar contracture.Methods 26 patients with scar contracture were randomly assigned into BTXA group and triamcinolone acetonide (TAC) group.Pinpoint tattooing was performed on each side of each scar in the plane of its longest axis.A template was used to ensure consistent length.These two tattoo points were measured to assess scar contraction at baseline,at every month for a total of 6 months.Histological analysis was conducted to study the physiological environment and immunohistochemistry to detect the expression of α-SMA and myosin-Ⅱ at different groups.Results Scar contraction was more relaxed in BTXA group than that in TAC group after 1 month (P<0.05),especially in the 6th month (the D value in BTXA group and TAC group was (1.23±0.42) cm,and (0.56±0.33) cm respectively).For immunohistochemistry,the expression of α-SMA and myosin-Ⅱ also decreased in BTXA group (P<0.05).Conclusions The treatment of scar contracture by suitable BTXA injections is safe and effective.%目的 探索A型肉毒毒素(botulinum toxin type A,BTXA)治疗挛缩性瘢痕的疗效.方法 选取26例挛缩性瘢痕患者,随机分为A型肉毒毒素组(BTXA组)和曲安奈德组(TAC组,对照组),注射药物治疗前测量各组患者瘢痕长轴长度,并于注射后再次测量其长度,1次/月,共6次,通过比较治疗前后差值评价药物疗效.切取各组瘢痕组织行免疫组织化学检测,观察α平滑肌肌动蛋白(α-SMA)及肌球蛋白-Ⅱ的表达情况.结果 药物作用1个月后,BTXA组较TAC组瘢痕挛缩程度明显减轻(P<0.05),尤以6个月时差异最明显,BTXA组和TAC组瘢痕长轴长度差值分别为(1.23±0.42) cm和(0.56±0.33) cm.免疫组织化学结果显示,BTXA组瘢痕内α-SMA及肌球蛋白-Ⅱ表达较TAC组明显减少(P<0.05).结论 A型肉毒毒素治疗挛缩性瘢痕操作简单、效果明显,值得推广应用.

  4. Noncosmetic periocular therapeutic applications of botulinum toxin

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    Kaynak-Hekimhan Pelin

    2010-01-01

    Full Text Available Botulinum toxin blocks acetylcholine release at the neuromuscular junction. The drug which was initially found to be useful in the treatment of strabismus has been extremely effective in the treatment of variety of conditions, both cosmetic and noncosmetic. Some of the noncosmetic uses of botulinum toxin applications include treatment of spastic facial dystonias, temporary treatment of idiopathic or thyroid dysfunction-induced upper eyelid retraction, suppression of undesired hyperlacrimation, induction of temporary ptosis by chemodenervation in facial paralysis, and correction of lower eyelid spastic entropion. Additional periocular uses include control of synchronic eyelid and extraocular muscle movements after aberrant regeneration of cranial nerve palsies. Cosmetic effects of botulinum toxin were discovered accidentally during treatments of facial dystonias. Some of the emerging nonperiocular application for the drug includes treatment of hyperhidrosis, migraine, tension-type headaches, and paralytic spasticity. Some of the undesired side effects of periocular applications of botulinum toxin inlcude ecchymosis, rash, hematoma, headache, flu-like symptoms, nausea, dizziness, loss of facial expression, lower eyelid laxity, dermatochalasis, ectropion, epiphora, eyebrow and eyelid ptosis, lagophthalmos, keratitis sicca, and diplopia.

  5. Application progress of botulinum toxin type A in the nervous system diseases%A型肉毒毒素在神经系统疾病中的应用研究进展

    Institute of Scientific and Technical Information of China (English)

    江伟; 何传斌

    2011-01-01

    In recent years, with the deepening study on the mechanism of botulinum toxin type A, the range of its applica -tions in the nervous system diseases continues to expand. Because botulinum toxin type A produces partial chemical dener-vation of the muscle resulting in a localized reduction in muscle activity, it can be used to treat hemifacial spasm, idio -pathic blepharospasm, oromandibular dystonia, spasmodic torticollis, tic disorder, limb spasticity after stroke and spastic cerebral palsy. In addition, it has recently begun to show some benefit in the control of hyperhidrosis ang excessive saliva -tion disease on account of its effects of the autonomic nervous system. It may also prove useful in treatment of a variety of headache and neuralgia owing to its analgesic effects. Therefore, botulinum toxin type A has become an important treatment option in the field of neurology.%近年来随着A型肉毒毒素作用机制研究的不断深入,其在神经系统疾病中的应用范围日渐扩大.因其肌肉化学性去神经支配效应可用于治疗面肌痉挛、特发性眼睑痉挛、口下颌肌张力障碍、痉挛性斜颈、抽动障碍、脑卒中后肢体痉挛和痉挛性脑瘫;因其植物神经系统效应可用于治疗多涎病症和多汗证;因其镇痛效应可用于治疗各种头痛和神经痛,故A型肉毒毒素已逐渐成为神经科领域一种重要的治疗手段.

  6. An Open Study of Botulinum-A Toxin Treatment of Idiopathic Trigeminal Neuralgia

    Directory of Open Access Journals (Sweden)

    Karim Nikkhah

    2015-07-01

    Full Text Available Introduction: Trigeminal Neuralgia (TN is a unilateral, recurrent, sharp facial pain disorder that is limited to the distribution of divisions of the trigeminal nerve. The aim of this study was to evaluate the efficacy of Botulinum neurotoxin type A (BTX-A for alleviating the frequency and severity of TN pain. Materials and Methods: This trial was performed as a before and after study. We treated 31 patients (15 male and 16 female with mean age of 52 year old that their diagnosis was made at least 4.5 years before. We injected BTX-A in various parts of face and particularly in the origin of mandibular and maxillary branches of trigeminal nerve. Injection volume was determined by the necessity and pain intensity measured with visual analog scale up to 100U. Patients were evaluated before and after the injection and were followed after week, and each month, for a three months period. Other related variables were recorded such as: toxin complications, pain status variations by brushing, chewing, cold weather and patient’s satisfaction with their therapy. Results: showed that after injection, pain intensity and frequency decreased after tooth brushing, chewing and cold weather (P

  7. 肉毒毒素B在儿童脑性瘫痪中的应用%Application of Botulinum Toxin B in Children with Cerebral Palsy

    Institute of Scientific and Technical Information of China (English)

    王雅洁

    2012-01-01

    肉毒毒素(BTX)是由肉毒菌素产生的一种细菌外毒素,是已知最毒的微生物毒素之一.BTX-B是由B型厌氧梭状肉毒芽胞杆菌Bean株产生的神经毒素,BTX-B不干扰内化、易位过程,在细胞基质裂解N-乙基-马来酰亚胺-敏感因子-附着蛋白受体复合物中的囊泡相关膜蛋白,干扰突触囊泡锚靠、融合,从而导致肌肉化学性失神经,降低肌肉痉挛.BTX-B还可适用于对BTX-A无反应以及不随意肌肌紧张的患儿.%Botulinum toxin( BTX )is a protein and neurotoxin produced by the bacterium clostridium botulinum, which is one of the most potent toxins known. BTX acts by blocking neurotransmitter release at the neuromuscular junction. BTX-B is produced by fermentation of the bacterium clostridium botulinum type B ( Bean strain). BTX-B does not interfere intemalization, translocation process, but cleaves synaptobrevin, which is a part of a protein complex necessary for proper docking and fusion. BTX-B blocks nerve activity in the muscles, causing a temporary reduction in muscle activity. BTX-B can be used as an effective and powerful medication. Its efficacy extends to patients who are resistant to BTX-A,and it can be used to treat involuntary muscle contractions.

  8. “Non-Toxic” Proteins of the Botulinum Toxin Complex Exert In-vivo Toxicity

    Science.gov (United States)

    Miyashita, Shin-Ichiro; Sagane, Yoshimasa; Suzuki, Tomonori; Matsumoto, Takashi; Niwa, Koichi; Watanabe, Toshihiro

    2016-01-01

    The botulinum neurotoxin (BoNT) causes muscle paralysis and is the most potent toxin in nature. BoNT is associated with a complex of auxiliary “Non-Toxic” proteins, which constitute a large-sized toxin complex (L-TC). However, here we report that the “Non-Toxic” complex of serotype D botulinum L-TC, when administered to rats, exerts in-vivo toxicity on small-intestinal villi. Moreover, Serotype C and D of the “Non-Toxic” complex, but not BoNT, induced vacuole-formation in a rat intestinal epithelial cell line (IEC-6), resulting in cell death. Our results suggest that the vacuole was formed in a manner distinct from the mechanism by which Helicobacter pylori vacuolating toxin (VacA) and Vibrio cholerae haemolysin induce vacuolation. We therefore hypothesise that the serotype C and D botulinum toxin complex is a functional hybrid of the neurotoxin and vacuolating toxin (VT) which arose from horizontal gene transfer from an ancestral BoNT-producing bacterium to a hypothetical VT-producing bacterium. PMID:27507612

  9. "Non-Toxic" Proteins of the Botulinum Toxin Complex Exert In-vivo Toxicity.

    Science.gov (United States)

    Miyashita, Shin-Ichiro; Sagane, Yoshimasa; Suzuki, Tomonori; Matsumoto, Takashi; Niwa, Koichi; Watanabe, Toshihiro

    2016-01-01

    The botulinum neurotoxin (BoNT) causes muscle paralysis and is the most potent toxin in nature. BoNT is associated with a complex of auxiliary "Non-Toxic" proteins, which constitute a large-sized toxin complex (L-TC). However, here we report that the "Non-Toxic" complex of serotype D botulinum L-TC, when administered to rats, exerts in-vivo toxicity on small-intestinal villi. Moreover, Serotype C and D of the "Non-Toxic" complex, but not BoNT, induced vacuole-formation in a rat intestinal epithelial cell line (IEC-6), resulting in cell death. Our results suggest that the vacuole was formed in a manner distinct from the mechanism by which Helicobacter pylori vacuolating toxin (VacA) and Vibrio cholerae haemolysin induce vacuolation. We therefore hypothesise that the serotype C and D botulinum toxin complex is a functional hybrid of the neurotoxin and vacuolating toxin (VT) which arose from horizontal gene transfer from an ancestral BoNT-producing bacterium to a hypothetical VT-producing bacterium. PMID:27507612

  10. Botulinum Toxin Treatment of Neuropathic Pain.

    Science.gov (United States)

    Mittal, Shivam Om; Safarpour, Delaram; Jabbari, Bahman

    2016-02-01

    Neuropathic pain (NP), a common form of human pain, often poorly responds to analgesic medications. In this review the authors discuss the pathophysiology and conventional treatment of neuropathic pain and provide evidenced-based statements on the efficacy of botulinum neurotoxins (BoNTs) in this form of pain. The level of efficacy for BoNT treatment in each category of NP is defined according to the published guidelines of the American Academy of Neurology. The data indicate that BoNT treatment (most of the literature is with onabotulinumtoxinA) is effective (level A evidence) in postherpetic neuralgia and trigeminal neuralgia. It is probably effective (level B) in posttraumatic neuralgia and painful diabetic neuropathy. The data on complex regional pain syndrome, carpal tunnel syndrome, occipital neuralgia, and phantom limb pain are preliminary and await conduction of randomized, blinded clinical trials. Much remains to be learned about the most-effective dosage and technique of injection, optimum dilutions, and differences among BoNTs in the treatment of neuropathic pain. PMID:26866499

  11. 研究应用A型肉毒毒素进行面部除皱的临床疗效%Clinical Efficacy of Application of Botulinum Toxin Type A on Facial Wrinkles

    Institute of Scientific and Technical Information of China (English)

    郜泽誉; 朱艳; 张志磊; 赵鹏

    2014-01-01

    目的:探讨和研究应用A型肉毒毒素进行面部除皱的临床治疗效果。方法:选取新疆医科大学第五附属医院2012年12月~2013年12月间接收的48例进行面部除皱的患者作为观察对象,对患者使用A型肉毒毒素进行注射治疗,每次使用A型肉毒毒素的总剂量不超过100U。结果:48例患者通过使用A型肉毒毒素注射之后,达到了很好的治疗效果,其中有46例患者的治疗效果显著有效,显著有效率为95.65%,治疗有效的为3例患者,有效率为6.25%,总有效率达到了100%。结论:使用A型肉毒毒素来去除面部的皱纹,其疗效非常显著,见效时间短,副作用小,患者的痛苦较少,是一种非常安全和有效的除皱方式,值得在临床上推广及应用。%Objective:To investigate the application and research of botulinum toxin type A in clinical treatment of facial wrinkles. Method:48 cases with facial wrinkles in the Fifth Affiliated Hospital of Xinjiang Medical University from December,2012 to December,2013 were regarded as research object. They were treated with botulinum toxin A injection. The total dose per use of botulinum toxin type A does not exceed 100U. Result:48 patients through the use of botulinum toxin type A injection achieved good therapeutic effect. The treatment effect of 46 cases was significantly effective,and the significant efficiency was 95.65%. 3 patients had effective treatment,the effective rate was 6.25%,the total effective rate was 100%. Conclusion:The effect of botulinum toxin type A is significant,the effective time is short,the side effect is little, and the pain of patients is less. So it is a very safe and effective anti wrinkle method,and worth application in clinical practice.

  12. A型肉毒毒素注射治疗腋臭疗效探讨%Effect of Botulinum Toxin type A injection on axillary bromidrosis

    Institute of Scientific and Technical Information of China (English)

    茅付勇

    2015-01-01

    Objective To investigate the clinical value of Botulinum Toxin A on axillary bromidrosis by minimally invasive injection. Methods Using minimally invasive Botulinum Toxin A injection method in the 60 cases of axillary bromidrosis (all of them are bilateral axillary), and then observing the curative effect. Results Of all the 60 patients 58 cases were followed up from March 2011 to June 2014,on average one year and a half,in order to observe the effect and complications and therapeutic effect of them,and the follow- up rate was 96.7% .The curative effect is satisfied. Conclusion Minimally invasive Botulinum Toxin A injection,as a non-operation therapy for axillary bromidrosis,is a method which is simple to operation,easy to master,region without scar,and the curative effect is satisfactory.%目的:探讨微创A型肉毒毒素注射法治疗腋臭的应用价值。方法:采用微创A型肉毒毒素注射法治疗腋臭60例(均为双侧腋窝)观察治疗效果。结果:60例患者中,随访58例,随访率为96.7%。随访时间2011年3月-2014年6月,平均1.5年,疗效满意。结论:微创A型肉毒毒素注射法治疗腋臭操作简单、易于掌握、创区无瘢痕,且疗效满意。

  13. A型肉毒毒素联合糖皮质激素治疗良性咬肌肥厚%Treatment of masseter hypertrophy with botulinum toxin type A and glucocorticoids

    Institute of Scientific and Technical Information of China (English)

    潘德玲; 袁继龙; 肖明

    2013-01-01

    Objective To investigate the clinical therapeutic effect of botulinum toxin type A and glu-cocorticoide on benign masseter hypertrophy.Methods Forty-seven cases ( 89 sides) with masseter hypertrophy were treated by the botulinum toxin type with 50 U per side and triamcinolone with 10 to 20 mg per side, the masseter thickness measured by CT and complications and adverse reactions were recorded at different time after injection.Results Compared with preoperative masseter thickness before injection, they was reduced significantly at different time after injection ( P<0.001).Totally 47 patients were satisfied with few complications.Conclusion Botulinum toxin type A and glucocorticoids injection is a safety, effective method for masseter hypertrophy.Prostecdtive efficacy should take a further observation.%目的 评价A型肉毒毒素联合糖皮质激素治疗良性咬肌肥大的疗效.方法 联合应用A型肉毒毒素和曲安奈德对47例(89侧)咬肌肥大求美者进行治疗,每一侧注射A型肉毒毒素剂量为50U、曲安奈德剂量为10~20mg,CT测量治疗前后不同时期求美者的咬肌厚度,记录不良反应和并发症.结果 在治疗后不同时期,47例(89侧)求美者的咬肌治疗区厚度均较治疗前减小(P <0.001),效果满意;47例均无严重并发症发生.结论 A型肉毒毒素联合糖皮质激素治疗咬肌肥大是一种安全有效的治疗方法,远期疗效尚需进一步观察.

  14. 16S rRNA in identification and typing of Clostridium botulinum%16S rRNA在肉毒梭菌分型鉴定中的应用

    Institute of Scientific and Technical Information of China (English)

    卢卫嘉; 毛晓燕; 熊颖; 张超; 王建锋

    2011-01-01

    Objective To establish a rapid, reliable method for identifying and typing of Clostridium botulinum using 16S rDNA sequencing and phylogenetic tree construction. Methods Clostridium genome templates were extracted from 9 strains, including LCL063, 830110, LC175, LCL155, 66418, N153, 61082, ALASKA, and IWANAIs 16S rRNA genes were amplifed by PCR with the 16S rRNA specific primers, and then the PCR products were cloned to pGEM -T Easy vector and sequenced. Finally, the sequence of 16S rDNA was analyzed by Clustal and Mega program; the phylogenetic trees were constructed by Neighor joining and Maximum parsimony. Results It was found that LCL063, 830110, and LCL175 were BONT/E producing Clostridium butyricums; IWANAI and ALASKA were Clostridium botulinum type E. The results of the present method were consistent with those of the conventional method. Conclusion 16S rRNA sequencing combined with phylogenetic tree analysis is a rapid and accurate method in Clostridium botulinum identification, and the method may serve as a criterion for bacterial typing with the completion of ribosomal RNA data bank.%目的 建立一种快速、可靠的对肉毒梭菌进行分型鉴定的手段.方法 以从LCL063、830110、LC175、LCL155、66418、N153、61082、ALASKA、IWANAI共9株梭菌中提取的基因组DNA为模板,利用16S rRNA特异性引物分别进行PCR扩增并进行T-A克隆转化、测序.通过Clustal和Mega软件分析16S rDNA序列,以NJ法和MP法构建进化树,分析其种属特异性.结果 16S rRNA分型结果可判断出LCL063、830110、LCL175为产E型毒素的酪酸梭菌.IWANAI与ALASKA株为E型肉毒梭菌.与传统分型鉴定得到的结果一致.结论 16S rRNA在肉毒梭菌分型鉴定中具有快速、准确的优势,随着核糖体库的不断完善,有望成为细菌分型鉴定的标准依据.

  15. A critical appraisal of the evidence for botulinum toxin type A in the treatment for cervico-thoracic myofascial pain syndrome.

    Science.gov (United States)

    Desai, Mehul J; Shkolnikova, Tatyana; Nava, Andrew; Inwald, Danielle

    2014-02-01

    Myofascial pain syndrome (MPS) is a musculoskeletal condition characterized by regional pain and muscle tenderness associated with the presence of myofascial trigger points (MTrPs). The last decade has seen an exponential increase in the use of botulinum toxin (BTX) to treat MPS. To understand the medical evidence substantiating the role of therapeutic BTX injections and to provide useful information for the medical practitioner, we applied the principles of evidence-based medicine to the treatment for cervico-thoracic MPS. A search was conducted through MEDLINE (PubMed, OVID, MDConsult), EMBASE, SCOPUS and the Cochrane database for the period 1966 to 2012 using the following keywords: myofascial pain, muscle pain, botulinum toxin, trigger points, and injections. A total of 7 trials satisfied our inclusion criteria and were evaluated in this review. Although the majority of studies found negative results, our analysis identified Gobel et al.'s as the highest quality study among these prospectively randomized investigations. This was due to appropriate identification of diagnostic criteria, excellent study design and objective endpoints. The 6 other identified studies had significant failings due to deficiencies in 1 or more major criteria. We conclude that higher quality, rigorously standardized studies are needed to more appropriately investigate this promising treatment modality.

  16. Botulinum Toxin A for Bladder Pain Syndrome/Interstitial Cystitis

    Science.gov (United States)

    Chiu, Bin; Tai, Huai-Ching; Chung, Shiu-Dong; Birder, Lori A.

    2016-01-01

    Botulinum neurotoxin A (BoNT-A), derived from Clostridium botulinum, has been used clinically for several diseases or syndrome including chronic migraine, spasticity, focal dystonia and other neuropathic pain. Chronic pelvic or bladder pain is the one of the core symptoms of bladder pain syndrome/interstitial cystitis (BPS/IC). However, in the field of urology, chronic bladder or pelvic pain is often difficult to eradicate by oral medications or bladder instillation therapy. We are looking for new treatment modality to improve bladder pain or associated urinary symptoms such as frequency and urgency for patients with BPS/IC. Recent studies investigating the mechanism of the antinociceptive effects of BoNT A suggest that it can inhibit the release of peripheral neurotransmitters and inflammatory mediators from sensory nerves. In this review, we will examine the evidence supporting the use of BoNTs in bladder pain from basic science models and review the clinical studies on therapeutic applications of BoNT for BPS/IC. PMID:27376330

  17. Observation of Curative Effect to Type A Botulinum Toxin for Treatment of Hypertrophy of the Masseter%A型肉毒素治疗咬肌肥大25例疗效观察

    Institute of Scientific and Technical Information of China (English)

    朱倩; 黄巡

    2006-01-01

    目的:寻求保守治疗咬肌肥大的有效方法.方法:采用A型肉毒素(botulinum toxin type-A,BTX-A)对25例咬肌肥大的患者进行治疗,每一侧注射剂量为50U.结果:25例治疗均有效,其中4例出现不对称,经过追加调整剂量后,效果满意,25例均无并发症.结论:A型肉毒素治疗咬肌肥大是一种安全有效的保守方法,远期疗效尚需进一步观察.

  18. Overview of Botulinum Toxins for Aesthetic Uses.

    Science.gov (United States)

    Gart, Michael S; Gutowski, Karol A

    2016-07-01

    Botulinum toxin type A (BTA) can be used for facial aesthetics. The 3 currently available BTA types include onabotulinumtoxinA (Botox; Botox Cosmetic, Allergan, Irvine, CA), abobotulinumtoxinA (Dysport; Ipsen, Ltd, Berkshire, UK), and incobotulinumtoxinA (Xeomin; Merz Pharmaceuticals, Frankfurt, Germany). The mechanism of action and clinical uses for treatment of dynamic lines of the forehead, brow, glabella, lateral orbit, nose, and lips are presented, as well as treatment of masseter hypertrophy, platysmal bands, and improvements of the perioral region. Specific BTA injection sites and suggested doses are presented. PMID:27363760

  19. TYPE A BOTULINUM TOXIN IN COMBINATION WITH HYALURONIC ACID IN EYE WEEK YOUNGER CURATIVE EFFECT OBSERVATION OF TREATMENT%A型肉毒素联合透明质酸在眼周年轻化的疗效观察

    Institute of Scientific and Technical Information of China (English)

    赵冰冰

    2015-01-01

    Objective To observe the type A botulinum toxin in eye week younger treatment combined the effect of hyaluronic acid. SelectionMethods Our hospital from March 2013 to March 2013 were 78 patients with facial wrinkles, and duration were observed.Results 78 cases of patients with, and follow-up of patients after half a year has 100% efficient. Conclusion Type A botulinum toxin in combination with hyaluronic acid for the treatment of facial wrinkles, and has less traumatic, had A higher safety, suitable for popularization and application in clinic.%目的:观察在眼周年轻化治疗中A型肉毒素联合透明质酸的效果。方法选取我院在2013年3月至2015年3月收治的面部皱纹患者78例,患者都给予A型肉毒素联合透明质酸进行注射治疗,对治疗后的效果、不良反应发生率及持续时间进行观察。结果78例患者的皱纹处在治疗后改善明显,并且在半年后对患者进行随访,具有100%的有效率。结论 A型肉毒素联合透明质酸对于治疗面部动静态皱纹,具有微创、安全、快捷,适合在有面部年轻化愿望的求美者中推广应用。

  20. A型肉毒毒素治疗Meige综合征的方法及效果分析%Botulinum toxin type A treatment of Meige syndrome of the method and result analysis

    Institute of Scientific and Technical Information of China (English)

    汪健; 高平峰; 周发明; 席刚明

    2011-01-01

    Objective Discusses botulinum toxin type A treatment of Meige syndrome the method and the effect.Methods 47 patients under the guidance of the EMG needle electrode with a double as the syringe,Spasm sites in patients with botulinum toxin type A intramuscular injection,each injected volume 0.1 ~0.2mi (including botulinum toxin 2.5~5U),number of injection points to 8 points points.According to Cohen,Albert spasm intensity grading evaluation curative effect.Results After the injection,the general 3~4days of effect,the curative effect continued for 3~6 months,the recrudescence duplicate injection was still effective,After the treatment,the patient myospasm intensity obvious drop, the comparison has the extremely significance difference before the treatment (P<0.01).The symptom alleviates and the obvious alleviation completely reaches 89.4% ,around the treatment the curative effect comparison has the extremely significance difference (P<0.01).Adverse reactions are mainly: local edema,bite muscle weakness,ptosis and so on,generally all can recover within two weeks.Conclusions Local injection of botulinum toxin type A treatment of Meige syndrome is a safe and effective and feasible treatment.%目的:探讨A型肉毒毒素治疗Meige综合征的方法及疗效.方法:47例患者在肌电图引导下用兼作注射器的针电极进行,在患者肌痉挛部位用A型肉毒毒素肌肉内注射,每点注射量为0.1~0.2ml(含肉毒毒素2.5~5 U),注射点数为数点~8点.根据Cohen、Albert痉挛强度分级评估疗效.结果:注射后一般3~4天起效,疗效持续3~6个月,复发者重复注射仍有效.治疗后患者肌痉挛强度明显下降,与治疗前比较有极显著性差异(P<0.01);症状完全缓解和明显缓解者达89.4%,治疗前后疗效比较有极显著性差异(P<0.01).不良反应主要有:局部水肿、咬肌无力、眼睑下垂等,一般两周内均能恢复.结论:局部注射A型肉毒毒素治疗Meige综合征为

  1. Botulinum toxin assessment, intervention and aftercare for paediatric and adult niche indications including pain: international consensus statement.

    Science.gov (United States)

    Rawicki, B; Sheean, G; Fung, V S C; Goldsmith, S; Morgan, C; Novak, I

    2010-08-01

    Evidence is emerging for the use of botulinum neurotoxin type-A (BoNT-A) for niche indications including pain independent of spasticity. Pain indications such as chronic nociceptive back pain, piriformis syndrome, chronic myofascial pain, pelvic pain, complex regional pain syndrome, facial pain and neuropathic pain are outlined in this paper. Of these, class I evidence is available for the treatment of chronic nociceptive low back pain, piriformis syndrome, myofascial pain, facial pain, neuropathic pain and plantar fasciitis. Peri-operative use of BoNT-A is emerging, with indications including planning for surgery and facilitating surgery, as well as healing and improving analgesia post-operatively. Evidence is limited, although there are some reports that clinicians are successfully using BoNT-A peri-operatively. There is class I evidence showing pre-operative use of BoNT-A has a beneficial effect on outcomes following adductor-release surgery. The use of BoNT for treatment of tremor, other than neck tremor in the setting of cervical dystonia, including evidence for upper limb tremor, cranial tremor and non-dystonic neck tremor is reviewed. The evidence is variable at this stage, and further study is required to develop definitive recommendations for the clinical utility of BoNT-A for these indications.

  2. Role of tetanus neurotoxin insensitive vesicle-associated membrane protein (TI-VAMP) in vesicular transport mediating neurite outgrowth.

    Science.gov (United States)

    Martinez-Arca, S; Alberts, P; Zahraoui, A; Louvard, D; Galli, T

    2000-05-15

    How vesicular transport participates in neurite outgrowth is still poorly understood. Neurite outgrowth is not sensitive to tetanus neurotoxin thus does not involve synaptobrevin-mediated vesicular transport to the plasma membrane of neurons. Tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI-VAMP) is a vesicle-SNARE (soluble N-ethylmaleimide-sensitive fusion protein [NSF] attachment protein [SNAP] receptor), involved in transport to the apical plasma membrane in epithelial cells, a tetanus neurotoxin-resistant pathway. Here we show that TI-VAMP is essential for vesicular transport-mediating neurite outgrowth in staurosporine-differentiated PC12 cells. The NH(2)-terminal domain, which precedes the SNARE motif of TI-VAMP, inhibits the association of TI-VAMP with synaptosome-associated protein of 25 kD (SNAP25). Expression of this domain inhibits neurite outgrowth as potently as Botulinum neurotoxin E, which cleaves SNAP25. In contrast, expression of the NH(2)-terminal deletion mutant of TI-VAMP increases SNARE complex formation and strongly stimulates neurite outgrowth. These results provide the first functional evidence for the role of TI-VAMP in neurite outgrowth and point to its NH(2)-terminal domain as a key regulator in this process.

  3. A study on the toxigenesis by Clostridium botulinum in nitrate and nitrite-reduced dry fermented sausages.

    Science.gov (United States)

    Hospital, Xavier F; Hierro, Eva; Stringer, Sandra; Fernández, Manuela

    2016-02-01

    Nitrite has been traditionally used to control Clostridium botulinum in cured meat products. However, in the case of dry fermented sausages, environmental factors such as pH, aw and the competitive microbiota may exert a more relevant role than nitrite in the inhibition of the growth and toxin production by C. botulinum. In this challenge test study, two varieties of Mediterranean dry sausages (salchichón and fuet) were inoculated with spores of C. botulinum Group I (proteolytic) and C. botulinum Group II (nonproteolytic). Sausages were prepared with 150 mg/kg of NaNO3 and 150 mg/kg of NaNO2 (maximum ingoing amounts allowed by the European Union regulation), with a 25% and 50% reduction, and without nitrate/nitrite. The initial pH in both products was 5.6, and decreased to values below 5.0 in salchichón and to 5.2 in fuet. Lactic acid bacteria counts reached 8-9 log cfu/g after fermentation. The aw decreased from initial values of 0.96 to about 0.88-0.90 at the end of ripening. Botulinum neurotoxin was not detected in any of the sausages, including those manufactured without nitrate and nitrite. Despite the environmental conditions were within the range for germination and growth of C. botulinum Group I during the first 8 days of the ripening process in fuet and 10-12 days in salchichón, acidity, aw and incubation temperature combined to inhibit the production of toxin, independently of the concentration of curing agents. Although decreasing or even removing nitrate/nitrite from the formula did not compromise safety regarding C. botulinum in the conditions tested in this study, their antimicrobial role should not be underestimated in the case that other hurdles could fail or other ripening conditions were used, and also considering the effect of nitrite on other pathogens. PMID:26619314

  4. Intramural injection with botulinum toxin significantly elongates the pig esophagus

    DEFF Research Database (Denmark)

    Larsen, Heidi Fhær; Jensen, Thorbjørn Søren Rønn; Rasmussen, Lars;

    2013-01-01

    Surgical treatment of long-gap esophageal atresia (LGEA) is challenging. Methods which facilitate stretching of the esophageal pouches may allow primary anastomosis. Botulinum toxin type A (BTX-A) blocks acetylcholine release in neuromuscular junctions, thereby causing muscle relaxation. We hypot...

  5. Botulinum toxin type A plus rehabilitative training for improving the motor function of the upper limbs and activities of daily life in patients with stroke and brain injury

    Institute of Scientific and Technical Information of China (English)

    Fei Guo; Wei Yue; Li Ren; Yumiao Zhang; Jing Yang

    2006-01-01

    BACKGROUND: Botulinum toxin type A (BTX-A) is mostly to be used to treat various diseases of motor disorders, whereas its effect on muscle spasm after stroke and brain injury needs further observation.OBJECTIVE : To observe the effect of BTX-A plus rehabilitative training on treating muscle spasm after stroke and brain injury.DESIGN: A randomized controlled observation.SETTINGS: Department of Rehabilitation, Department of Neurology and Department of Neurosurgery, the Second Hospital of Hebei Medical University.PARTICIPANTS: Sixty inpatients with brain injury and stroke were selected from the Department of Rehabilitation, Department of Neurology and Department of Neurosurgery, the Second Hospital of Hebei Medical University from January 2001 to August 2006. They were all confirmed by CT and MRI, and had obvious increase of spastic muscle strength in upper limbs, their Ashworth grades were grade 2 or above. The patients were randomly divided into treatment group (n =30) and control group (n =30).METHODS: ① Patients in the treatment group undertook comprehensive rehabilitative trainings, and they were administrated with domestic BTX-A, which was provided by Lanzhou Institute of Biological Products, Ministry of Health (S10970037), and the muscles of flexion spasm were selected for upper limbs, 20-25 IU for each site.② Patients in the treatment group were assessed before injection and at 1 and 2 weeks, 1 and 3 months after injection respectively, and those in the control group were assessed at corresponding time points. The recovery of muscle spasm was assessed by modified Ashworth scale (MAS, grade 0-Ⅳ; Grade 0 for without increase of muscle strength; Grade Ⅳ for rigidity at passive flexion and extension); The recovery of motor function of the upper limbs was evaluated with Fugl-Meyer Assessment (FMA, total score was 226 points, including 100 for exercise, 14 for balance, 24 for sense, 44 for joint motion, 44 for pain and 66 for upper limb); The ADL were

  6. Substrate Recognition Mechanism of VAMP/Synaptobrevin-cleaving Clostridial Neurotoxins*S⃞

    Science.gov (United States)

    Sikorra, Stefan; Henke, Tina; Galli, Thierry; Binz, Thomas

    2008-01-01

    Botulinum neurotoxins (BoNTs) and tetanus neurotoxin (TeNT) inhibit neurotransmitter release by proteolyzing a single peptide bond in one of the three soluble N-ethylmaleimide-sensitive factor attachment protein receptors SNAP-25, syntaxin, and vesicle-associated membrane protein (VAMP)/synaptobrevin. TeNT and BoNT/B, D, F, and G of the seven known BoNTs cleave the synaptic vesicle protein VAMP/synaptobrevin. Except for BoNT/B and TeNT, they cleave unique peptide bonds, and prior work suggested that different substrate segments are required for the interaction of each toxin. Although the mode of SNAP-25 cleavage by BoNT/A and E has recently been studied in detail, the mechanism of VAMP/synaptobrevin proteolysis is fragmentary. Here, we report the determination of all substrate residues that are involved in the interaction with BoNT/B, D, and F and TeNT by means of systematic mutagenesis of VAMP/synaptobrevin. For each of the toxins, three or more residues clustered at an N-terminal site remote from the respective scissile bond are identified that affect solely substrate binding. These exosites exhibit different sizes and distances to the scissile peptide bonds for each neurotoxin. Substrate segments C-terminal of the cleavage site (P4-P4′) do not play a role in the catalytic process. Mutation of residues in the proximity of the scissile bond exclusively affects the turnover number; however, the importance of individual positions at the cleavage sites varied for each toxin. The data show that, similar to the SNAP-25 proteolyzing BoNT/A and E, VAMP/synaptobrevin-specific clostridial neurotoxins also initiate substrate interaction, employing an exosite located N-terminal of the scissile peptide bond. PMID:18511418

  7. A型肉毒杆菌毒素的壳聚糖/海藻酸钠缓释微囊研制和药物的体外释放速率研究%The Study In Vitro Release Rate Study of Botulinum Toxin type A Chitosan/Alginate Microcapsules Using Research and Drugs

    Institute of Scientific and Technical Information of China (English)

    吴河坪; 林小铭; 曾明兵; 陈静嫦

    2015-01-01

    目的:研制包裹A型肉毒杆菌毒素的壳聚糖/海藻酸钠微囊,并测定其体外药物释放动力学。方法:采用注射器滴注法制成包裹A型肉毒杆菌毒素的壳聚糖/海藻酸钠微囊混悬液。采用高效液相色谱法测定A型肉毒杆菌毒素的标准曲线,每隔2周测定微囊所包裹的A型肉毒杆菌毒素体外释放浓度。结果:微囊形态为圆形透明颗粒状,粒径在1mm左右。用SPSS统计软件进行分析微囊中A型肉毒杆菌毒素的体外每2周释放量,显示药物累积释放量与时间之间为直线关系,基本符合药物零级释放动力学。结论:滴注法制备包裹A型肉毒杆菌毒素的壳聚糖/海藻酸钠缓释微囊制作方法简便,其药物体外释放稳定。%Objective:To develop parcels of botulinum toxin type A chitosan/alginate and tested for in vitro drug release kinetics. Method:using a syringe infusion into the legal package botulinum toxin type A chitosan/alginate suspension. Determination of botu-linum A toxin standard curve, two weeks entrapped microcapsules measured botulinum toxin type A in vitro release every concentra-tion using high performance liquid chromatography. Results: The microcapsules form a circular transparent granular diameter of about 1mm. Using SPSS statistical software for analysis microcapsules A botulinum toxin in vitro release every two weeks, show cumu-lative drug release and the time between a linear relationship, in line with the zero-order release kinetics of drugs. Conclusion:chito-san infusion prepared package of botulinum toxin type A/alginate microcapsules using the production method is simple, its in vitro drug release and stability.

  8. Tratamiento de la cefalea tensional con toxina botulínica tipo A Botulinum toxin type A for the treatment of tension-type headache

    Directory of Open Access Journals (Sweden)

    D. Arbaiza

    2006-06-01

    el área total de aplicación, como probables factores en la respuesta analgésica.Objectives: To determine the time of onset of the analgesia, the < ficacy and the security of the application of the botulinu toxin A (BTA in patient with tension-type headache (TH. Material and methods: Inclusion criteria: patients between 18 and 70 yeai diagnosis of TH according to the International Headache S ciety, two or more episodes of TH per month during at lea 3 months, non controlled TH with the standard treatmei normal neurological examination and signed inform consei Variables analyzed: Intensity of headache (IH, frequency presentation of headache (FH, duration of headache (DH sensibility to digit pressure (SDP of the cranial musculatur number of painful points (NPD of the cranial musculatur time of onset of the analgesia (TBA, time until the ma: mum effect of analgesia (TMA and decrease of use of an.gesics (DUA. Evaluations in the days 0, 15 and 30. Fifty units of BT were applied to cranial musculature. The patients could continue with their previous analgesic therapy according to the evolution of the headache. Results: Twenty female patients were included with a median age of 47 years. IH 2.15 in the first evaluation diminished at 1.05 and 0.9 in the second and third evaluations (p=0.009 and 0.008. FH of 10.25 days diminished to 3.6 and 2.75 days (p=0.008 and 0.012. DH of 2.3 daily hours it diminished at 1.3 and 1.1 (p=0.009 in both. SDP 1.55 diminished at 0.5 and 0.55 (p=0.008 in both. NPD 2.65 diminished at 0.8 and 0.6 (p=0.010 and 0.006. TBA was 8 days and TMA was 25 days. DUA 5 diminished at 1.9 and 1.5. Around of 35% of the patients presented a probable adverse event of mild or moderate severity. Conclusion: TB-TO it is a simple, effective treatment and provides control of the TH. The technique is easy to reproduce. More studies are needed to value the number of applications and the total area of application, as probable factors in the analgesic response.

  9. Prevalence of toxin-producing Clostridium botulinum associated with the macroalga Cladophora in three Great Lakes: growth and management.

    Science.gov (United States)

    Lan Chun, Chan; Kahn, Chase I; Borchert, Andrew J; Byappanahalli, Muruleedhara N; Whitman, Richard L; Peller, Julie; Pier, Christina; Lin, Guangyun; Johnson, Eric A; Sadowsky, Michael J

    2015-04-01

    The reemergence of avian botulism caused by Clostridium botulinum type E has been observed across the Great Lakes in recent years. Evidence suggests an association between the nuisance algae, Cladophora spp., and C. botulinum in nearshore areas of the Great Lakes. However, the nature of the association between Cladophora and C. botulinum is not fully understood due, in part, to the complex food web interactions in this disease etiology. In this study, we extensively evaluated their association by quantitatively examining population size and serotypes of C. botulinum in algal mats collected from wide geographic areas in lakes Michigan, Ontario, and Erie in 2011-2012 and comparing them with frequencies in other matrices such as sand and water. A high prevalence (96%) of C. botulinum type E was observed in Cladophora mats collected from shorelines of the Great Lakes in 2012. Among the algae samples containing detectable C. botulinum, the population size of C. Botulinum type E was 10(0)-10(4) MPN/g dried algae, which was much greater (up to 10(3) fold) than that found in sand or the water column, indicating that Cladophora mats are sources of this pathogen. Mouse toxinantitoxin bioassays confirmed that the putative C. botulinum belonged to the type E serotype. Steam treatment was effective in reducing or eliminating C. botulinum type E viable cells in Cladophora mats, thereby breaking the potential transmission route of toxin up to the food chain. Consequently, our data suggest that steam treatment incorporated with a beach cleaning machine may be an effective treatment of Cladophora-borne C. botulinum and may reduce bird mortality and human health risks.

  10. Prevalence of toxin-producing Clostridium botulinum associated with the macroalga Cladophora in three Great Lakes: growth and management

    Science.gov (United States)

    Chun, Chan Lan; Kahn, Chase I.; Borchert, Andrew J.; Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Peller, Julie R.; Pier, Christina; Lin, Guangyun; Johnson, Eric A.; Sadowsky, Michael J.

    2015-01-01

    The reemergence of avian botulism caused by Clostridium botulinum type E has been observed across the Great Lakes in recent years. Evidence suggests an association between the nuisance algae, Cladophoraspp., and C. botulinum in nearshore areas of the Great Lakes. However, the nature of the association between Cladophora and C. botulinum is not fully understood due, in part, to the complex food web interactions in this disease etiology. In this study, we extensively evaluated their association by quantitatively examining population size and serotypes of C. botulinum in algal mats collected from wide geographic areas in lakes Michigan, Ontario, and Erie in 2011–2012 and comparing them with frequencies in other matrices such as sand and water. A high prevalence (96%) of C. botulinum type E was observed inCladophora mats collected from shorelines of the Great Lakes in 2012. Among the algae samples containing detectable C. botulinum, the population size of C. Botulinum type E was 100–104 MPN/g dried algae, which was much greater (up to 103 fold) than that found in sand or the water column, indicating thatCladophora mats are sources of this pathogen. Mouse toxinantitoxin bioassays confirmed that the putativeC. botulinum belonged to the type E serotype. Steam treatment was effective in reducing or eliminating C. botulinum type E viable cells in Cladophora mats, thereby breaking the potential transmission route of toxin up to the food chain. Consequently, our data suggest that steam treatment incorporated with a beach cleaning machine may be an effective treatment of Cladophora-borne C. botulinum and may reduce bird mortality and human health risks.

  11. Botulinum toxin injection improved voluntary motor control in selected patients with post-stroke spasticity

    Institute of Scientific and Technical Information of China (English)

    Shuo-Hsiu Chang; Gerald E Francisco; Sheng Li

    2012-01-01

    The effect of botulinum toxin type A injection on voluntary grip control was examined in a 53-year-old female, who sustained a hemorrhagic right middle cerebral artery stroke 3 years previously, which resulted in finger flexor spasticity and residual weak finger/wrist extension. The patient received 50 units of botulinum toxin type A injection each to the motor points (2 sites/muscle) of the left flexor digitorum superficialis and flexor digitorum profundus, respectively. Botulinum toxin injection led to weakness and tone reduction in the spastic finger flexors, but improved grip release time in grip initiation/release reaction time tasks. Improved release time was accompanied by shortened extensor electromyography activity, and improved release time likely correlated with blocked co-contraction of finger flexors during voluntary finger extension. This case report demonstrated that botulinum toxin injection improved voluntary motor control of the hand in a chronic stroke patient with residual finger extension.

  12. Safety Evaluation of Sous Vide-Processed Products with Respect to Nonproteolytic Clostridium botulinum by Use of Challenge Studies and Predictive Microbiological Models

    Science.gov (United States)

    Hyytiä-Trees, Eija; Skyttä, Eija; Mokkila, Mirja; Kinnunen, Arvo; Lindström, Miia; Lähteenmäki, Liisa; Ahvenainen, Raija; Korkeala, Hannu

    2000-01-01

    Sixteen different types of sous vide-processed products were evaluated for safety with respect to nonproteolytic group II Clostridium botulinum by using challenge tests with low (2.0-log-CFU/kg) and high (5.3-log-CFU/kg) inocula and two currently available predictive microbiological models, Food MicroModel (FMM) and Pathogen Modeling Program (PMP). After thermal processing, the products were stored at 4 and 8°C and examined for the presence of botulinal spores and neurotoxin on the sell-by date and 7 days after the sell-by date. Most of the thermal processes were found to be inadequate for eliminating spores, even in low-inoculum samples. Only 2 of the 16 products were found to be negative for botulinal spores and neurotoxin at both sampling times. Two products at the high inoculum level showed toxigenesis during storage at 8°C, one of them at the sell-by date. The predictions generated by both the FMM thermal death model and the FMM and PMP growth models were found to be inconsistent with the observed results in a majority of the challenges. The inaccurate predictions were caused by the limited number and range of the controlling factors in the models. Based on this study, it was concluded that the safety of sous vide products needs to be carefully evaluated product by product. Time-temperature combinations used in thermal treatments should be reevaluated to increase the efficiency of processing, and the use of additional antibotulinal hurdles, such as biopreservatives, should be assessed. PMID:10618228

  13. Safety evaluation of sous vide-processed products with respect to nonproteolytic Clostridium botulinum by use of challenge studies and predictive microbiological models.

    Science.gov (United States)

    Hyytiä-Trees, E; Skyttä, E; Mokkila, M; Kinnunen, A; Lindström, M; Lähteenmäki, L; Ahvenainen, R; Korkeala, H

    2000-01-01

    Sixteen different types of sous vide-processed products were evaluated for safety with respect to nonproteolytic group II Clostridium botulinum by using challenge tests with low (2. 0-log-CFU/kg) and high (5.3-log-CFU/kg) inocula and two currently available predictive microbiological models, Food MicroModel (FMM) and Pathogen Modeling Program (PMP). After thermal processing, the products were stored at 4 and 8 degrees C and examined for the presence of botulinal spores and neurotoxin on the sell-by date and 7 days after the sell-by date. Most of the thermal processes were found to be inadequate for eliminating spores, even in low-inoculum samples. Only 2 of the 16 products were found to be negative for botulinal spores and neurotoxin at both sampling times. Two products at the high inoculum level showed toxigenesis during storage at 8 degrees C, one of them at the sell-by date. The predictions generated by both the FMM thermal death model and the FMM and PMP growth models were found to be inconsistent with the observed results in a majority of the challenges. The inaccurate predictions were caused by the limited number and range of the controlling factors in the models. Based on this study, it was concluded that the safety of sous vide products needs to be carefully evaluated product by product. Time-temperature combinations used in thermal treatments should be reevaluated to increase the efficiency of processing, and the use of additional antibotulinal hurdles, such as biopreservatives, should be assessed. PMID:10618228

  14. Botulinum Toxin Injections: A Treatment for Muscle Spasms

    Science.gov (United States)

    ... A Treatment for Muscle Spasms What is botulinum toxin? Botulinum toxin is a protein that helps stop muscle ... won't have any harmful effects from the toxin. Botulinum toxin has been used safely for a number ...

  15. Prevention and treatment of early hypertrophic scars with botulinum toxin type A%A型肉毒毒素预防与治疗早期增生性瘢痕的临床初探

    Institute of Scientific and Technical Information of China (English)

    于波; 陈敏亮; 刘文阁; 徐永成; 孙同柱

    2008-01-01

    Objective To explore the effect of botulinum toxin type A(BTXA)in the treatment of early hypertrophic scars(HTS).Methods BTXA was injected into and around the eady HTS,and then the modal and histological changes of the scars as well as the clinical reaction were observed in the patient.BTXA was also injected into muscle around the incision and effect on the cicatrization observed.Results Injection of BTXA could obviously alleviate ache and pruritus of eady HTS and could impel the atrophy and inteneration of eady HTS.Changes were found in paraffin-embedded tissue section by the hemetoxylin and eosin(HE)staining.Injection of BTXA into muscle around the cut could can reduce occurrence of HTS.Conclusion BTXA can help prevent the early HTS to a certain extent.The mechanism underlying this effect may be related to the reducing the tension around scars and proliferative activity,interfering with the signal transduction of small nerves,affecting the proliferation and apoptosis of fibroblasts and subsequently decreasing the collagen synthesis.%目的 探索A型肉毒毒素(botulinum toxin type A,BTXA)对早期增生性瘢痕(hypertrophic scar,HTS)的临床预防及治疗效果.方法 早期HTS周围及组织内注射BTXA,观察瘢痕注射药物后形态学变化、组织学变化及临床症状表现;手术切口缝合后即刻向周围肌肉浅层注射BTXA,观察远期瘢痕愈合情况.结果 局部注射BTXA可以明显减轻早期HTS疼痛和瘙痒症状,促使瘢痕组织萎缩、软化;组织切片HE染色显示HTS组织内结构有所变化.同时,手术切口周围肌肉浅层注射BTXA可以降低术后切口HTS的发生、发展概率.结论 BTXA对早期HTS具有一定程度的治疗和预防作用,其疗效可能是通过降低瘢痕两侧张力及活动,干扰瘢痕内小神经传导,以及影响成纤维细胞增殖分化,促进凋亡进而减少胶原合成而起作用.

  16. INJECTION BOTULINUM IN PARALYTIC STRABISMUS

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    Krishna Kishore

    2015-09-01

    Full Text Available AIM: To assess the effectiveness of botulinum toxin injection to the antagonist muscle in paralytic strabismus. OBJECTIVE: To study the effect of botulinum toxin a injection into the antagonist muscle in cases of paralytic strabismus to alleviate diplopia. MATERIAL AND METHODS: This was a tertiary eye care hospital based prospective interventional study in the department of Orthoptics over a period from October 2011 to October 2013. 36 patients with paralytic strabismus of recent onset within 3 months, with chief complaint of double vision were included. RESULTS: The study data analysis of 36 patients of paralytic strabismus of recent onset (within 3 months with chief complaints of double vision showed a ge wise distribution as 3(8.33% in 0 - 20 years, 16(44.44% in 21 - 40 years, 16(44.44% in 41 - 60 years, 1(2.78% > 60 years. g ender wise 26(72.22% males, 10 (27.28% females, a etiology wise 15(41.67% were diabetic, 4(11.11% were traumatic, 11(30.56% were Id iopathic, 2(5.56% were due to CSOM and 4(11.11% due to Diabetes and Hypertension. All patients were treated with botulinum toxin injection to the antagonist nonoperatic muscle and were followed at an interval of 1 week, 1 month and 3 months. Thorough cli nical examination and Diplopia charting were done before and after treatment. CONCLUSION: Injection botulinum into the antagonist muscle during the first three months after the onset allows the patients to enjoy and appreciate fusion in primary gaze without necessity for head turn, Prevents contracture of antagonist muscle. Thus botulinum toxin is useful in the treatment of acute paretic loss of ocular muscle function when surgical treatment of the ocular muscles is not yet possible but the patient is obviously disturbed by diplopia or forced head posture. The procedure is simple, safe and effective method.

  17. Efficacy and safety of a single botulinum type A toxin complex treatment (Dysport) for the relief of upper back myofascial pain syndrome: results from a randomized double-blind placebo-controlled multicentre study.

    Science.gov (United States)

    Göbel, Hartmut; Heinze, Axel; Reichel, Gerhard; Hefter, Harald; Benecke, Reiner

    2006-11-01

    Botulinum type A toxin (BoNT-A) has antinociceptive and muscle-relaxant properties and may help relieve the symptoms of myofascial pain syndrome. In this study we evaluated the efficacy and tolerability of BoNT-A (Dysport) in patients with myofascial pain syndrome of the upper back. We conducted a prospective, randomized, double-blind, placebo-controlled, 12-week, multicentre study. Patients with moderate-to-severe myofascial pain syndrome affecting cervical and/or shoulder muscles (10 trigger points, disease duration 6-24 months) were randomized to Dysport or saline. Injections were made into the 10 most tender trigger points (40 units per site). The primary outcome was the proportion of patients with mild or no pain at week 5. Secondary outcomes included changes in pain intensity and the number of pain-free days per week. Tolerability and safety were also assessed. At week 5, significantly more patients in the Dysport group reported mild or no pain (51%), compared with the patients in the placebo group (26%; p=0.002). Compared with placebo, Dysport resulted in a significantly greater change from baseline in pain intensity during weeks 5-8 (ppain between weeks 5 and 12 (p=0.036). Treatment was well tolerated, with most side effects resolving within 8 weeks. In conclusion, in patients with upper back myofascial pain syndrome, injections of 400 Ipsen units of Dysport at 10 individualised trigger points significantly improved pain levels 4-6 weeks after treatment. Injections were well tolerated.

  18. Evaluating Functional Outcomes of Botulinum Toxin Type A Injection Combined with Occupational Therapy in the Upper Limbs of Children with Cerebral Palsy: A 9-Month Follow-Up from the Perspectives of Both Child and Caregiver.

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    Yu-Ching Lin

    Full Text Available To assess the effectiveness of combining botulinum toxin type A (BoNT-A with functional occupational therapy (OT at 9-month follow-up in children with cerebral palsy (CP with bilateral upper limb impairments from the perspectives of both child and caregiver.Twelve children with CP and their caregivers were assessed across 5 time points over 9 months based on the ICF after BoNT-A injection and functional OT in this open-label study.Significant differences were found across the 5 time points (p < .05 for both grasp and visual-motor integration with small effects (effect sizes = 0.12-0.24 and the self-care capability and performance of social function (p < .05. However, based on the effect sizes (0.02-0.14, no significant effects were found at the 4 post-test time points. Small effects were found on the psychological domain (effect sizes = 0.25-0.37 and environmental domains (effect size = 0.27 at follow-ups.Combining a BoNT-A injection with OT not only reduced the muscle tone and increased ROM but also improved the upper limb function and self-care capability in children with CP. More importantly, these effects persisted for up to 9 months. Functional OT extends the effectiveness of a BoNT-A injection.

  19. Reconstituting botulinum toxin drugs: shaking, stirring or what?

    Science.gov (United States)

    Dressler, Dirk; Bigalke, Hans

    2016-05-01

    Most botulinum toxin (BT) drugs are stored as powders which need to be reconstituted with normal saline before clinical use. As botulinum neurotoxin (BNT), the therapeutically active ingredient, is a large double-stranded protein the process of reconstitution should be performed with special attention to mechanical stress applied. We wanted to test the mechanical stability of BNT during the reconstitution process. For this, 100 MU onabotulinumtoxinA (Botox(®), Irvine, CA, USA) was reconstituted with 2.0 ml of NaCl/H2O. Gentle reconstitution (GR) was performed with a 5 ml syringe, a 0.90 × 70 mm injection needle, one cycle of injection-aspiration-injection and two gentle shakes of the vial. Aggressive reconstitution (AR) was performed with a 5 ml syringe, a 0.40 × 40 mm injection needle, ten injection-aspiration-injection cycles and 30 s of continuous shaking of the vial. AR increased the time to paralysis in the mouse hemidiaphragm assay (HDA) from 72.0 ± 4.6 to 106.0 ± 16.0 min (*p = 0.002, two-tailed t test after Kolmogorov-Smirnova test with Lilliefors correction for normal distribution). Construction of a calibration curve revealed that the increase in the time to paralysis was correlated with a loss of potency of from 100 to 58 MU (-42 %). BT users should use large diameter injection needles for reconstitution, apply two or three injection-aspiration-injection cycles and, maybe, shake the vials a few times to rinse the entire glass wall. Aggressive reconstitution with small diameter needles, prolonged injection-aspiration-injection and violent shaking should be avoided.

  20. Effects of Botulinum Toxin Type A on Cell Apoptosis in Rat Benign Prostatic Hyperplasia Model%A型肉毒毒素注射引起大鼠前列腺增生模型细胞凋亡的研究

    Institute of Scientific and Technical Information of China (English)

    杨雄; 杨栋; 侯腾; 马彦; 汪隆旺; 韩晓敏; 李兵; 陈朝晖

    2011-01-01

    目的:探讨A型肉毒毒素注射后前列腺的组织形态学变化.方法:建立SD大鼠前列腺增生模型,模型大鼠前列腺内分别注射不同剂量的A型肉毒毒素后,TUNEL检测前列腺细胞的凋亡,免疫组织化学技术检测对照组与10 U肉毒毒素注射组Bcl-2、Fas及Caspase-3的表达差异.结果:随着注射剂量的加大,大鼠前列腺体积及湿重出现明显下降,细胞凋亡增加.相比对照组,10U肉毒毒素注射组Bcl-2的表达下调而Fas及Caspase-3的表达上调.结论:A型肉毒毒素能有效的缩小前列腺体积,引起前列腺细胞的凋亡.%Objective:To investigate the morphological changes of rat prostate after the injection of Botulinum toxin type A. Methods: We established prostatic hyperplasia model of SD rats and injected different doses of botulinum toxin type A into the prostates of BPH rat model. Prostatic cells apoptosis was detected by TUNEL assay.The expression of Bcl-2、Fas, and Caspase-3 were evaluated as well. Results:In the botulinum toxin type A injection group, it was observed that the volume and wet weight of rat prostates were decreased, that cell apoptosis was increased. The expressions of Bcl-2, Fas, and Caspase-3 in 10U botulinum toxin injection group were different compared to the control group. Conclusions: Botulinum toxin type A can diminish the volume and wet weight of rat prostate and induce apoptosis of prostatic cells.

  1. Diversity of Group I and II Clostridium botulinum Strains from France Including Recently Identified Subtypes

    Science.gov (United States)

    Mazuet, Christelle; Legeay, Christine; Sautereau, Jean; Ma, Laurence; Bouchier, Christiane; Bouvet, Philippe; Popoff, Michel R.

    2016-01-01

    In France, human botulism is mainly food-borne intoxication, whereas infant botulism is rare. A total of 99 group I and II Clostridium botulinum strains including 59 type A (12 historical isolates [1947–1961], 43 from France [1986–2013], 3 from other countries, and 1 collection strain), 31 type B (3 historical, 23 recent isolates, 4 from other countries, and 1 collection strain), and 9 type E (5 historical, 3 isolates, and 1 collection strain) were investigated by botulinum locus gene sequencing and multilocus sequence typing analysis. Historical C. botulinum A strains mainly belonged to subtype A1 and sequence type (ST) 1, whereas recent strains exhibited a wide genetic diversity: subtype A1 in orfX or ha locus, A1(B), A1(F), A2, A2b2, A5(B2′) A5(B3′), as well as the recently identified A7 and A8 subtypes, and were distributed into 25 STs. Clostridium botulinum A1(B) was the most frequent subtype from food-borne botulism and food. Group I C. botulinum type B in France were mainly subtype B2 (14 out of 20 historical and recent strains) and were divided into 19 STs. Food-borne botulism resulting from ham consumption during the recent period was due to group II C. botulinum B4. Type E botulism is rare in France, 5 historical and 1 recent strains were subtype E3. A subtype E12 was recently identified from an unusual ham contamination. Clostridium botulinum strains from human botulism in France showed a wide genetic diversity and seems to result not from a single evolutionary lineage but from multiple and independent genetic rearrangements. PMID:27189984

  2. [Use of botulinum toxin in strabismus].

    Science.gov (United States)

    Wabbels, B

    2016-07-01

    Botulinum toxin can be a useful tool for treating acute sixth nerve palsy and excessive eye deviations due to unstable Graves' disease, when surgery is not yet possible. The diagnostic injection for estimation of possible postoperative double vision also makes sense. In convergence spasms, periocular botulinum toxin injections can be a therapeutic option. Botulinum toxin is not a first line option in infantile esotropia without binocularity or in adult horizontal strabismus. Side effects include ptosis and vertical deviations. PMID:27369733

  3. Toxina botunílica tipo B no manejo de distonia não-responsiva a toxina botunílica tipo A Botulinum toxin type B in the management of dystonia non-responsive to botulinum toxin type A

    Directory of Open Access Journals (Sweden)

    Francisco Cardoso

    2003-09-01

    sessões de 17,4 semanas (variação 16-18. Ela se tornou não-respondedora após a nona sessão. Primeiro tratamento com BTX-B, 6000U, recebeu escore 0. A segunda sessão, 12000U, produziu escore 4.Paciente 4- Com a idade de 69 anos este homem desenvolveu distonia cranial idiopática. Antes de ser atendido por mim, ele recebeu 6 sessões de BTX-A em outros serviços. Na minha instituição ele foi tratado com dose acumulada de 730U em 4 sessões com intervalo médio entre sessões de 16,3 semanas (variação 15-18, havendo perdido a resposta na sexta sessão. Tratamento com BTX-B, 12000U, produziu escore 4 e durou 20 semanas. Efeitos colaterais: dor local (todos os pacientes e ressecamento da boca e ptose palpebral (um paciente cada. CONCLUSÃO: Meus achados confirmam que injeções de BTX-B são eficazes e seguras para pacientes não-respondedores secundários a BTX-A. Os resultados também mostram ser necessário individualizar a dose de BTX-B para se alcançar melhores resultados.BACKGROUND: Botulinum toxin (BTX injection is the first choice treatment for focal dystonias. However 10% or more of patients who receive repetitive injections of BTX type A (BTX-A lose response (secondary non-responders. One of the strategies to manage such patients is to treat them with another serotype. The aim of this article is to describe my experience with BTX type B (BTX-B in the management of patients with focal dystonia who became secondary non-responders to BTX-A. METHOD: Open-label non-controlled use of BTX-B injections to treat dystonia patients who developed secondary nonresponse to BTX-A Response to treatment was rated on a 0-4 scale (Jankovic. RESULTS: Four patients entered the study. Pacient 1- At age 48 this man developed idiopathic cervical dystonia. Five years later he also presented with blepharospasm and idiopathic oromandibular dystonia. He was treated with 7604U of BTX-A along 23 sessions separated by a mean interval of 18.8 weeks (range 6-39. Loss of response was

  4. The use of botulinum toxin as primary or adjunctive treatment for post acne and traumatic scarring

    Directory of Open Access Journals (Sweden)

    Greg J Goodman

    2010-01-01

    Full Text Available Background : Botulinum toxin has been utilised successfully in many facial and extra facial regions to limit superfluous movement. Scars, whether traumatic or disease-related, are treated with many modalities. Objective: To assess the available literature concerning the prophylactic use of botulinum toxin for the improvement in the cosmetic outcome of scars induced by surgery and to examine its role in the treatment of established scars alone, as also combined with other modalities. Material and Methods : The results of the prophylactic use of botulinum toxin to limit the resultant scarring from surgery are examined by a literature review. The primary and adjunctive use of botulinum toxin in the treatment of post acne and post surgical and traumatic scars is explored by case examples. Results : Literature review and personal experience shows good Improvement in the appearance of scars with the use of botulinum toxin alone or with other adjuvant modalities in the treatment of scars. Conclusion : Botulinum toxin would appear to be useful both in the prophylaxis and treatment of certain types of scars.

  5. Outbreak of Type C Botulism in Commercial Layer Chickens.

    Science.gov (United States)

    Sato, Yuko; Wigle, William L; Gallagher, Susan; Johnson, Amy L; Sweeney, Raymond W; Wakenell, Patricia S

    2016-03-01

    This report describes an outbreak of type C botulism in two organic, free-range commercial layer farms in the Midwest. Hens affected were 64-wk-old Hy-Line brown hens and 34-wk-old Hy-Line brown hens owned by the same company, but housed on different premises, with approximately 20,000 birds per house. Mortality over the 2 wk of investigation was estimated to be up to 8% and 2.8%, respectively, with birds acting listless, lethargic, and depressed. Clinical signs consisted of progressive paralysis, and severely affected birds were moribund and laterally recumbent. Hens had ruffled feathers that easily epilated, with loss of muscular tone in the neck, tail, and wings. Hens had closed eyes and were reluctant to move. There were no significant gross or histopathologic lesions. Intestinal samples were submitted to the University of Pennsylvania Botulism Diagnostic Laboratory for real-time PCR and were positive for Clostridium botulinum organisms containing the Type C neurotoxin gene. Speculations on the source of the botulinum toxins include poor mortality removal leading to cannibalism of decomposing carcasses, as well as birds on the farm having access to putrid carcasses in the compost pile from a hole in their outdoor access fence. PMID:26953952

  6. A型肉毒毒素治疗痉挛型脑瘫的研究进展%Research progress of botulinum toxin type A in treatment of spastic cerebral palsy

    Institute of Scientific and Technical Information of China (English)

    刘春明(综述); 刘芸(审校)

    2015-01-01

    Cerebral palsy refers to brain injury caused by various reasons from before birth to 1 month after birth or movement disorders and abnormal posture caused by brain developmental defects. Children cerebral palsy is divided into 5 kinds in the clinic:spastic type, dyskinetic type, ataxia type, muscle hypotonia type and mixed type. Among them, the spastic type accounts for 60% ~70% , and is the main type which causes children physical disabilities. Once diagnosed, the comprehensive treatment method is ap-plied, including functional training, physical therapy, acupuncture, massage, medicine treatment, and so on. In recent years, the lo-cal injection of botulinum toxin A is used for treating the children with spastic cerebral palsy. A large number of clinical studies show that the drug is safe and effective; however, in the course of treatment, it should strictly control the drug indications and contraindica-tions. Especially, the dosage choice and the accurate localization of target muscle is the key factor to influence the treatment effect.%脑瘫是指出生前到出生后1个月内各种原因所引起的脑损伤或发育缺陷所致的运动障碍及姿势异常。小儿脑瘫临床上分为痉挛型,不随意运动型,共济失调型,肌张力低下型及混合型5种,其中,痉挛型占60%~70%,是造成儿童肢体残疾的最主要脑瘫类型。脑瘫一旦确诊多采用综合的治疗方法包括功能训练,物理疗法,中医针刺,按摩,药物治疗等。近年来,临床上开始广泛使用毒杆菌毒素 A(botuli-num toxin A,BTX-A)局部注射治疗痉挛型脑瘫儿童,大量临床研究结果显示该药安全、有效,但在治疗过程中应该严格掌握使用该药的适应证,禁忌证,而剂量的选择和靶肌的准确定位是影响治疗效果的关键因素。

  7. Toxin yet not toxic: Botulinum toxin in dentistry.

    Science.gov (United States)

    Archana, M S

    2016-04-01

    Paracelsus contrasted poisons from nonpoisons, stating that "All things are poisons, and there is nothing that is harmless; the dose alone decides that something is a poison". Living organisms, such as plants, animals, and microorganisms, constitute a huge source of pharmaceutically useful medicines and toxins. Depending on their source, toxins can be categorized as phytotoxins, mycotoxins, or zootoxins, which include venoms and bacterial toxins. Any toxin can be harmful or beneficial. Within the last 100 years, the perception of botulinum neurotoxin (BTX) has evolved from that of a poison to a versatile clinical agent with various uses. BTX plays a key role in the management of many orofacial and dental disorders. Its indications are rapidly expanding, with ongoing trials for further applications. However, despite its clinical use, what BTX specifically does in each condition is still not clear. The main aim of this review is to describe some of the unclear aspects of this potentially useful agent, with a focus on the current research in dentistry. PMID:27486290

  8. Botulinum toxin injection techniques for the management of adult spasticity.

    Science.gov (United States)

    Walker, Heather W; Lee, Michael Y; Bahroo, Laxman B; Hedera, Peter; Charles, David

    2015-04-01

    Spasticity is often experienced by individuals with injury or illness of the central nervous system from etiologies such as stroke, spinal cord injury, brain injury, multiple sclerosis, or other neurologic conditions. Although spasticity may provide benefits in some patients, it more often leads to complications negatively impacting the patient. Nonpharmacologic treatment options often do not provide long-term reduction of spasticity, and systemic interventions, such as oral medications, can have intolerable side effects. The use of botulinum neurotoxin injections is one option for management of focal spasticity. Several localization techniques are available to physicians that allow for identification of the selected target muscles. These methods include anatomic localization in isolation or in conjunction with electromyography guidance, electrical stimulation guidance, or ultrasound guidance. This article will focus on further description of each of these techniques in relation to the treatment of adult spasticity and will discuss the advantages and disadvantages of each technique, as well as review the literature comparing the techniques. PMID:25305369

  9. A型肉毒素治疗痉挛型脑性瘫痪120例临床疗效分析%Clinical Efficacy Analysis of Botulinum Toxin Type A Treating 120 Cases of Spastic Cerebral Palsy

    Institute of Scientific and Technical Information of China (English)

    李华伟; 马丙祥

    2013-01-01

    Objective:To investigate the curative effects of injection with botulinum toxin type A (BTX-A) and rehabilitation therapy on spastic cere-bral palsy (CP), with location-guide of electrical stimulation. Methods:120 cases of children with spastic cerebral palsy received BTX-A injection and comprehensive rehabilitation treatment afterwards with electrical stimulation guided, to observe the changes of muscle tension and range of joint passive motion. Results:Muscle tension decreased significantly after the injection (P<0.01);range of joint passive motion significantly increased (P<0.05). Conclusion:BTX-A injection can significantly reduce muscle tension, enlarge range of joint motion and improve motor function in children with cerebral palsy.%  目的:探讨电刺激定位引导下A型肉毒毒素(BTX-A)注射结合康复治疗对痉挛性脑性瘫痪(CP)的治疗效果。方法:120例痉挛性脑瘫患儿均在电刺激仪引导下,进行BTX-A注射,注射后接受综合康复治疗,并观察其肌张力和关节被动活动度的变化。结果:注射后患儿的肌张力均明显降低(P<0.01);关节被动活动度均明显增大(P<0.05)。结论:BTX-A注射能显著降低脑瘫患儿的肌张力,扩大关节活动度,提高运动功能。

  10. Comparison of clinical efficacy between radiofrequency ablation and botulinum toxin type A in treatment of benign masseter hypertrophy%射频消融与A型肉毒毒素治疗咬肌良性肥大的临床效果比较

    Institute of Scientific and Technical Information of China (English)

    陈刚; 黄金龙; 张骏; 刘育凤; 闻可; 刘宁; 王金明

    2015-01-01

    Objective To evaluate masseter thickness and its complications of radiofrequency ablation and botulinum toxin type A in the treatment of masseter hypertrophy.Methods Totally 36 patients of benign masseter hypertrophy were randomly divided into radiofrequency ablation therapy group (18 patients) and the injection of botulinum toxin type A treatment group (18 patients).The thickness of the masseter and complications were analyzed before treatment,6 months and 12 months after treatment.Results The average thickness of masseter muscle in botulinum toxin type A and radiofrequency ablation groups was significantly reduced 6 or 12 months after treatment,and the difference was statistically significant (P<0.05).However,the thickness of masseter muscle in radiofrequency ablation group after 12 months reduced greater than that in botulinum toxin type A group (P<0.05).Complications of botulinum toxin type A group were swallow difficulty (one patient),unnatural facial expressions (4 patients) and pain at injection site (10 patients) and that in radiofrequency ablation treatment group were that 18 patients had swelling of the skin 1 to 2 weeks after treatment.All patients had recovery 2 weeks after treatment.There was no facial nerve,parotid duct injury,or oral dysfunction.Conclusions The lasting effect of radiofrequency ablation treatment for benign masseter hypertrophy is longer than that of botulinum toxin type A.%目的 比较射频消融和A型肉毒毒素注射治疗咬肌肥大的临床效果.方法 将36例咬肌良性肥大的患者按照治疗方法不同完全随机分为接受射频消融治疗组和注射A型肉毒毒素治疗组,每组18例.评价治疗前、治疗后6个月、12个月时咬肌的厚度及并发症.结果 A型肉毒杆菌毒素治疗组中,与治疗前相比,右侧或左侧平均咬肌厚度治疗后6、12个月明显减少,差异有统计学意义(P<0.05).射频消融治疗组中,与治疗前相比,治疗后6、12个月咬

  11. Botulinum toxin type A in refractory chronic migraine: an open-label trial Toxina botulínica tipo A no tratamento da enxaqueca refratária: um estudo aberto

    Directory of Open Access Journals (Sweden)

    Carla Menezes

    2007-09-01

    Full Text Available Botulinum toxin type A (BT-A has been described as an important strategy to various types of pain such as cervical dystonia, myofascial pain syndrome and headache. Although BT-A efficacy has not been proven in tension type headache, its use in migraine continues controversial. In this open trial, we evaluated the efficacy of BT-A in refractory migraine. BT-A was injected in patients diagnosed with migraine who had previously used three classes of prophylactic drugs by at least one year with no response. The most important improvement was observed within 30 days, but pain intensity and frequency of headache had been decreased until the end of three months of follow up. Side effects of BT-A were mild and self limited. We conclude that BT-A seems to be a safe and effective treatment to refractory migraine patients.Toxina botulínica tipo A (TB-A tem sido descrita como importante estratégia para diversos tipos de dor como cefaléia e dores relacionadas a distonia cervical ou síndrome miofascial. Embora a eficácia da TB-A não tenha sido demonstrada na cefaléia do tipo tensional, seu uso na enxaqueca continua controverso. Nesse estudo avaliamos a eficácia da TB-A na enxaqueca refratária. TB-A foi injetada em pacientes com enxaqueca que fizeram tratamento prévio com no mínimo três classes de medicamentos profiláticos, sem resultados satisfatórios. A melhora mais significativa dos pacientes foi observada após 30 dias de aplicação de TB-A, enquanto intensidade da dor e freqüência de cefaléia continuaram reduzidas até o final de três meses de seguimento. Os efeitos colaterais observados após a aplicação de TB-A foram moderados e auto-limitados. Os nossos dados mostram que TB-A parece ser um tratamento seguro e eficaz para pacientes com enxaqueca refratária.

  12. The Role of Botulinum Toxin A in the Treatment of Raynaud Phenomenon.

    Science.gov (United States)

    Segreto, Francesco; Marangi, Giovanni Francesco; Cerbone, Vincenzo; Persichetti, Paolo

    2016-09-01

    Raynaud phenomenon (RP) is a transient digital ischemia that occurs after exposure to cold temperature or emotional distress. It presents with a triphasic course: the initial white phase is followed by cyanotic discoloration and, subsequently, erythema. The attacks may be associated with pain, paresthesia, and complicate with nonhealing ulceration often leading to amputation. To date, there are no clear-cut therapeutic guidelines and many medications are used off-label. Encouraging results were reported with the use of botulinum neurotoxin-A (BoNT-A). However, there is still ongoing debate regarding indications, contraindications, best injection technique, and mechanism of action. The aim of this study was to address these issues by providing an up-to-date and detailed overview of the use of BoNT-A in RP.A PubMed database search was conducted. The available studies and techniques were evaluated and compared.The search yielded a total of 29 studies. Ten papers, published between 2004 and 2014, were considered relevant. A total of 128 patients underwent BoNT-A injections. Seventy-five percent to 100 % of the patients reported pain reduction after treatment. Healing of ulcers was reported in 75% to 100% of the affected patients. The most common complication was temporary hand weakness, with an average incidence of 14.1%. Injections targeting the neurovascular bundle at or slightly proximal to the A1 pulley were the most commonly performed.Botulinum neurotoxin-A injection proved to be a valid approach in both primary and secondary RP. The available evidence shows the achievement of both symptomatic and functional improvements in this debilitating condition. However, the patient should be adequately informed about the risk of transient hand weakness. PMID:26808752

  13. Characterization of the spore surface and exosporium proteins of Clostridium sporogenes; implications for Clostridium botulinum group I strains.

    Science.gov (United States)

    Janganan, Thamarai K; Mullin, Nic; Tzokov, Svetomir B; Stringer, Sandra; Fagan, Robert P; Hobbs, Jamie K; Moir, Anne; Bullough, Per A

    2016-10-01

    Clostridium sporogenes is a non-pathogenic close relative and surrogate for Group I (proteolytic) neurotoxin-producing Clostridium botulinum strains. The exosporium, the sac-like outermost layer of spores of these species, is likely to contribute to adhesion, dissemination, and virulence. A paracrystalline array, hairy nap, and several appendages were detected in the exosporium of C. sporogenes strain NCIMB 701792 by EM and AFM. The protein composition of purified exosporium was explored by LC-MS/MS of tryptic peptides from major individual SDS-PAGE-separated protein bands, and from bulk exosporium. Two high molecular weight protein bands both contained the same protein with a collagen-like repeat domain, the probable constituent of the hairy nap, as well as cysteine-rich proteins CsxA and CsxB. A third cysteine-rich protein (CsxC) was also identified. These three proteins are also encoded in C. botulinum Prevot 594, and homologues (75-100% amino acid identity) are encoded in many other Group I strains. This work provides the first insight into the likely composition and organization of the exosporium of Group I C. botulinum spores. PMID:27375261

  14. Conformational divergence in the HA-33/HA-17 trimer of serotype C and D botulinum toxin complex.

    Science.gov (United States)

    Sagane, Yoshimasa; Hayashi, Shintaro; Akiyama, Tomonori; Matsumoto, Takashi; Hasegawa, Kimiko; Yamano, Akihito; Suzuki, Tomonori; Niwa, Koichi; Watanabe, Toshihiro; Yajima, Shunsuke

    2016-08-01

    Clostridium botulinum produces a large toxin complex (L-TC) comprising botulinum neurotoxin associated with auxiliary nontoxic proteins. A complex of 33- and 17-kDa hemagglutinins (an HA-33/HA-17 trimer) enhances L-TC transport across the intestinal epithelial cell layer via binding HA-33 to a sugar on the cell surface. At least two subtypes of serotype C/D HA-33 exhibit differing preferences for the sugars sialic acid and galactose. Here, we compared the three-dimensional structures of the galactose-binding HA-33 and HA-33/HA-17 trimers produced by the C-Yoichi strain. Comparisons of serotype C/D HA-33 sequences reveal a variable region with relatively low sequence similarity across the C. botulinum strains; the variability of this region may influence the manner of sugar-recognition by HA-33. Crystal structures of sialic acid- and galactose-binding HA-33 are broadly similar in appearance. However, small-angle X-ray scattering revealed distinct solution structures for HA-33/HA-17 trimers. A structural change in the C-terminal variable region of HA-33 might cause a dramatic shift in the conformation and sugar-recognition mode of HA-33/HA-17 trimer. PMID:27237978

  15. [Botulinum toxin in disabling dermatological diseases].

    Science.gov (United States)

    Messikh, R; Atallah, L; Aubin, F; Humbert, P

    2009-05-01

    Botulinum toxin could represent nowadays a new treatment modality especially for cutaneous conditions in course of which conventional treatments remain unsuccessful. Besides palmar and plantar hyperhidrosis, botulinum toxin has demonstrated efficacy in different conditions associated with hyperhidrosis, such as dyshidrosis, multiple eccrine hidrocystomas, hidradenitis suppurativa, Frey syndrome, but also in different conditions worsened by hyperhidrosis such as Hailey-Hailey disease, Darier disease, inversed psoriasis, aquagenic palmoplantar keratoderma, pachyonychia congenital. Moreover, different cutaneous conditions associated with sensitive disorders and/or neurological involvements could benefit from botulinum toxin, for example anal fissures, leg ulcers, lichen simplex, notalgia paresthetica, vestibulitis. Endly, a case of cutis laxa was described where the patient was improved by cutaneous injections of botulinum toxin. PMID:19576479

  16. Vocal aging and adductor spasmodic dysphonia: Response to botulinum toxin injection

    Directory of Open Access Journals (Sweden)

    Michael P Cannito

    2008-03-01

    Full Text Available Michael P Cannito, Joel C Kahane, Lesya ChornaSchool of Audiology and Speech-Language Pathology, The University of Memphis, Memphis, TN, USAAbstract: Aging of the larynx is characterized by involutional changes which alter its biomechanical and neural properties and create a biological environment that is different from younger counterparts. Illustrative anatomical examples are presented. This natural, non-disease process appears to set conditions which may influence the effectiveness of botulinum toxin injection and our expectations for its success. Adductor spasmodic dysphonia, a type of laryngeal dystonia, is typically treated using botulinum toxin injections of the vocal folds in order to suppress adductory muscle spasms which are disruptive to production of speech and voice. A few studies have suggested diminished response to treatment in older patients with adductor spasmodic dysphonia. This retrospective study provides a reanalysis of existing pre-to-post treatment data as function of age. Perceptual judgments of speech produced by 42 patients with ADSD were made by two panels of professional listeners with expertise in voice or fluency of speech. Results demonstrate a markedly reduced positive response to botulinum toxin treatment in the older patients. Perceptual findings are further elucidated by means of acoustic spectrography. Literature on vocal aging is reviewed to provide a specific set of biological mechanisms that best account for the observed interaction of botulinum toxin treatment with advancing age.Keywords: vocal aging, adductor spasmodic dysphonia, botulinum toxin, voice quality, speech fluency

  17. The first non Clostridial botulinum-like toxin cleaves VAMP within the juxtamembrane domain

    Science.gov (United States)

    Zornetta, Irene; Azarnia Tehran, Domenico; Arrigoni, Giorgio; Anniballi, Fabrizio; Bano, Luca; Leka, Oneda; Zanotti, Giuseppe; Binz, Thomas; Montecucco, Cesare

    2016-01-01

    The genome of Weissella oryzae SG25T was recently sequenced and a botulinum neurotoxin (BoNT) like gene was identified by bioinformatics methods. The typical three-domains organization of BoNTs with a N-terminal metalloprotease domain, a translocation and a cell binding domains could be identified. The BoNT family of neurotoxins is rapidly growing, but this was the first indication of the possible expression of a BoNT toxin outside the Clostridium genus. We performed molecular modeling and dynamics simulations showing that the 50 kDa N-terminal domain folds very similarly to the metalloprotease domain of BoNT/B, whilst the binding part is different. However, neither the recombinant metalloprotease nor the binding domains showed cross-reactivity with the standard antisera that define the seven serotypes of BoNTs. We found that the purified Weissella metalloprotease cleaves VAMP at a single site untouched by the other VAMP-specific BoNTs. This site is a unique Trp-Trp peptide bond located within the juxtamembrane segment of VAMP which is essential for neurotransmitter release. Therefore, the present study identifies the first non-Clostridial BoNT-like metalloprotease that cleaves VAMP at a novel and relevant site and we propose to label it BoNT/Wo. PMID:27443638

  18. The first non Clostridial botulinum-like toxin cleaves VAMP within the juxtamembrane domain.

    Science.gov (United States)

    Zornetta, Irene; Azarnia Tehran, Domenico; Arrigoni, Giorgio; Anniballi, Fabrizio; Bano, Luca; Leka, Oneda; Zanotti, Giuseppe; Binz, Thomas; Montecucco, Cesare

    2016-07-22

    The genome of Weissella oryzae SG25T was recently sequenced and a botulinum neurotoxin (BoNT) like gene was identified by bioinformatics methods. The typical three-domains organization of BoNTs with a N-terminal metalloprotease domain, a translocation and a cell binding domains could be identified. The BoNT family of neurotoxins is rapidly growing, but this was the first indication of the possible expression of a BoNT toxin outside the Clostridium genus. We performed molecular modeling and dynamics simulations showing that the 50 kDa N-terminal domain folds very similarly to the metalloprotease domain of BoNT/B, whilst the binding part is different. However, neither the recombinant metalloprotease nor the binding domains showed cross-reactivity with the standard antisera that define the seven serotypes of BoNTs. We found that the purified Weissella metalloprotease cleaves VAMP at a single site untouched by the other VAMP-specific BoNTs. This site is a unique Trp-Trp peptide bond located within the juxtamembrane segment of VAMP which is essential for neurotransmitter release. Therefore, the present study identifies the first non-Clostridial BoNT-like metalloprotease that cleaves VAMP at a novel and relevant site and we propose to label it BoNT/Wo.

  19. Botulinum toxin type A in the treatment of hemifacial spasm: an 11-year experience Toxina botulínica tipo A no tratamento do espasmo hemifacial: 11 anos de experiência

    Directory of Open Access Journals (Sweden)

    Egberto Reis Barbosa

    2010-08-01

    Full Text Available In order to evaluate the long-term effect of botulinum toxin type A (BTX in the treatment of hemifacial spasm (HFS, a retrospective analysis of patients treated at the Movement Disorders Unit of the Division of Neurology, Clinical Hospital, University of São Paulo, School of Medicine from 1993 to 2004 was made. A total of 808 injections with BTX were administered to 54 patients with HFS. The mean duration of improvement per application was 3.46 months and the mean rate of improvement using subjective judgement by the patient was of 83%. Adverse effects, mostly minor, were observed in 64.8% of patients at least once along the period of follow-up and the most frequent of them was orbicularis oris paralysis (38.8%. There was no decrement in response when compared the first and the last injection recorded.Para avaliar o efeito em longo prazo da toxina botulínica tipo A (TXB no tratamento do espasmo hemifacial (EHF, foi feita uma análise retrospectiva de pacientes tratados no Ambulatório de Distúrbios do Movimento da Divisão de Clínica Neurológica - Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo no período de 1993 a 2004. Um total de 808 aplicações de TXB foram administradas a 54 pacientes com EHF. A duração média de melhora foi de 3,46 meses e a taxa média de melhora segundo avaliação subjetiva do paciente foi de 83%. Efeitos adversos, em sua maioria menores, foram observados em 64,8% dos pacientes ao menos uma vez durante o seguimento e o mais freqüente foi paralisia do orbicular da boca (38,3%. Não se observou decremento na resposta quando se comparou a primeira com a última aplicação anotada.

  20. Botulinum toxin type A for the treatment of palmar hyperhidrosis: a prospective study of 26 patients%A型肉毒毒素注射治疗26例手多汗症的临床观察

    Institute of Scientific and Technical Information of China (English)

    杨红军; 彭凯润; 邓兵梅; 程飚

    2012-01-01

    手多汗症是指发生于手掌的过多的出汗,给造成社交和就业上的困难.本研究对A型肉毒毒素治疗手多汗症的疗效进行临床观察.选择26例手多汗症患者(女17例,男9例),参照多汗症量表评估其出汗严重程度.记录患者A型肉毒毒素注射前后出汗评分、起效时间、副作用等指标.结果发现A型肉毒毒素注射后手出汗明显减少,效果持续3个月以上,而且副作用轻微且一过性.A型肉毒毒素注射治疗手多汗症是安全、有效的实用方法.%Palmar hyperhidrosis is characterized by excessive sweating in hand,it has a significant impact on the social and professional relationships of affected individuals.The aim of this study was to assess effect of botulinum toxin type A (BTX-A) to patients with palmar hyperhidrosis.The study included 26 patients (17 women and 9 men) with a diagnosis of Palmar hyperhidrosis treated with BTX-A. severity of hyperhidrosis qualitative measured by The Hyperhidrosis Disease Severity Scale (HDSS). All patients completed a questionnaire included the following information: time to effect of BTX-A; local or systemic side effects; and severity of hyperhidrosis before and after BTX-A treatment.There was a highly significant reduction in the severity of hyperhidrosis 3 months after performing the treatment with BTX-A. The side effects of treatment were sligtht and short. Treatment of Palmar hyperhidrosis with BTX-A was well tolerated and effective.

  1. A型肉毒毒素对脑性瘫痪患儿腘绳肌痉挛的疗效%Effect of Botulinum Toxin Type A on Spastic Hamstring in Children with Cerebral Palsy

    Institute of Scientific and Technical Information of China (English)

    刘建军; 席冰玉; 纪树荣; 吴卫红; 张雁; 曾凡勇; 李南玲

    2014-01-01

    目的:观察A型肉毒毒素(BTX-A)注射缓解脑瘫患儿腘绳肌痉挛的疗效。方法39例存在腘绳肌痉挛的脑瘫患儿分为对照组(n=20)和观察组(n=19),对照组采用常规康复训练,观察组在常规康复训练的基础上予BTX-A局部注射。治疗前后采用改良Ashworth量表(MAS)、粗大运动功能测试(GMFM-88)、膝关节屈曲角度进行评定。结果治疗6周后,观察组患儿腘绳肌MAS评分、膝关节屈曲角度、GMFM-88评分均较治疗前改善(P<0.05),且均优于对照组(P<0.05);对照组GMFM-88较治疗前改善(P<0.05)。结论 BTX-A注射可以有效缓解脑瘫患儿腘绳肌痉挛,有助于纠正异常步态,提高患儿运动功能。%Objective To observe the effect of Botulinum toxin type A (BTX-A) injection on spastic hamstring in the children with cere-bral palsy. Methods 39 cerebral palsy children with spastic hamstring were divided into control group (n=20) and experimental group (n=19). The control group accepted physical therapy, while the experimental group accepted BTX-A injection in affected hamstring in addition. They were assessed with modified Ashworth Scale (MAS), Gross Motor Fucntion Measure (GMFM-88) and flexion angle of knee joints be-fore and after treatment. Results The scores of MAS and GMFM-88, and flexion angle of knee joints improved significantly 6 weeks after treatment in the experimental group (P<0.05), and improved more than that in the control group (P<0.05). Only the score of GMFM-88 im-proved significantly in the control group (P<0.05). Conclusion BTX-A injection can relieve hamstring spasticity in children with cerebral palsy, which may be helpful to correct abnormal gait and improve the motor function.

  2. A型肉毒毒素对脑性瘫痪患儿腘绳肌痉挛的疗效%Effect of Botulinum Toxin Type A on Spastic Hamstring in Children with Cerebral Palsy

    Institute of Scientific and Technical Information of China (English)

    刘建军; 席冰玉; 纪树荣; 吴卫红; 张雁; 曾凡勇; 李南玲

    2014-01-01

    Objective To observe the effect of Botulinum toxin type A (BTX-A) injection on spastic hamstring in the children with cere-bral palsy. Methods 39 cerebral palsy children with spastic hamstring were divided into control group (n=20) and experimental group (n=19). The control group accepted physical therapy, while the experimental group accepted BTX-A injection in affected hamstring in addition. They were assessed with modified Ashworth Scale (MAS), Gross Motor Fucntion Measure (GMFM-88) and flexion angle of knee joints be-fore and after treatment. Results The scores of MAS and GMFM-88, and flexion angle of knee joints improved significantly 6 weeks after treatment in the experimental group (P<0.05), and improved more than that in the control group (P<0.05). Only the score of GMFM-88 im-proved significantly in the control group (P<0.05). Conclusion BTX-A injection can relieve hamstring spasticity in children with cerebral palsy, which may be helpful to correct abnormal gait and improve the motor function.%目的:观察A型肉毒毒素(BTX-A)注射缓解脑瘫患儿腘绳肌痉挛的疗效。方法39例存在腘绳肌痉挛的脑瘫患儿分为对照组(n=20)和观察组(n=19),对照组采用常规康复训练,观察组在常规康复训练的基础上予BTX-A局部注射。治疗前后采用改良Ashworth量表(MAS)、粗大运动功能测试(GMFM-88)、膝关节屈曲角度进行评定。结果治疗6周后,观察组患儿腘绳肌MAS评分、膝关节屈曲角度、GMFM-88评分均较治疗前改善(P<0.05),且均优于对照组(P<0.05);对照组GMFM-88较治疗前改善(P<0.05)。结论 BTX-A注射可以有效缓解脑瘫患儿腘绳肌痉挛,有助于纠正异常步态,提高患儿运动功能。

  3. Botulinum toxin treatment of hemifacial spasm.

    OpenAIRE

    Elston, J S

    1986-01-01

    Six patients with hemifacial spasm were treated with injections of botulinum toxin A into the orbicularis oculi; the abnormal movements around the eye were relieved for an average of 15 weeks. There were no systemic or significant local side effects, and in view of the risks involved in neurosurgical treatment, a trial of botulinum toxin injections is recommended in the first instance in this condition.

  4. Clinical studies of small amount of botulinum toxin type a local injection in patients with upper facial wrinkles%局部注射少量A型肉毒毒素治疗上半面部皱纹的临床研究

    Institute of Scientific and Technical Information of China (English)

    尚晓旭; 吴景东

    2011-01-01

    目的:观察局部注射少量A型肉毒毒素(BTXA)去除上半面部皱纹的临床疗效.方法:应用A型肉毒毒素的198例患者,其中眉间纹65例,鱼尾纹80例,额纹20例,鼻背部皱纹33例.所有患者行局部多点注射A型肉毒毒素,浓度为4U/0.1ml,并根据不同的患者调整剂量和浓度.记录患者疗效、维持时间与不良反应.结果:A型肉毒毒素治疗面部上三分之一皱纹有效率为100%,显效率94.4%.肌肉麻痹的效果通常持续3~6个月不等,不良反应主要为4例上睑下垂,3例局部水肿和4例青紫,各占2.0%、1.5%和2.0%.上述症状于3~10内自行完全消失.结论:A型肉毒毒素局部注射治疗面部上三分之一皱纹起效迅速、无创伤、简便易行.%Objective To observe the clinical effect of small amount of botulinum toxin type A (BTXA) local injection to remove half of the clinical effect of facial wrinkles. Methods 198 botulinum toxin type A patients, including 65 cases of frown lines, 80 cases of crow's feet, 20 cases of the wrinkles on the forehead, 33 cases of wrinkles of nasal back. All patients underwent local multi-point injection of botulinum toxin type A, the concentration is 4U/0.1ml, and adjusted the dose according to different patient and concentration. We recorded the outcomes, effective duration and adverse reactions. Results The effective rate of botulinum toxin type A in treatment with facial wrinkles was 100%, the obvious rate was 94.4%,and the effective duration lasts 3-6 months. The adverse effects of treatment were four cases of ptosis, 3 and 4 cases with local edema and bruising,taking account 2.0%, 1.5% and 2.0%, respectively. The symptoms disappeared on their own way. Conclusion A local injection of botulinum toxin type A treatment of upper a third facial wrinkles was rapid onset, non-invasive, non-traumatic and simple.

  5. 两种A型肉毒毒素在前额的弥散性比较%Diffuseness comparison of two kinds of botulinum toxins type A on the forehead

    Institute of Scientific and Technical Information of China (English)

    姜海燕; 陈淑君; 周珺

    2016-01-01

    Objective To evaluate the diffuse characteristics of two types of botulinum toxin type A (BOTOX and CBTX A) in the forehead and the effect of intradermal (ID) and subcutanous (SC) infections on their diffusion.Methods Healthy volunteers (n =20) were recruited to receive a 0.05 ml (2 U) injection of botulinum toxin type A at four forehead sites [both sides of medial forehead (SC) and temporal forehead (ID)],one side for BOTOX and another for CBTX-A.On day 14,the Minors' iodine starch test was performed and photographs were taken for calculating the area and dimensions of anhydrotic area.Results The areas of anhidrosis ID were significantly greater with CBTX-A (180±15) mm2 than that of BOTOX (144±14) mm2.The same conditions occurred in vertical dimensions (14.4±1.1) mm vs.(15.8±1.0) mm and horizontal dimensions (10.8±0.6) mm vs.(12.5±0.7) mm.There was significant variation between the both for diffuseness in the two anhidrotic halos observed after ID injection (P<0.05).The areas of anhidrosis by SC were significantly greater with CBTX-A (207±17) mm2 than that in BOTOX (183±18) mm2.The mean horizontal dimension was greater with CBTX-A by SC (13.2±0.6) mm than that of BOTOX by SC (12.2±0.7) mm,but not statistically significant (P =0.06).The mean horizontal dimension in BOTOX was significantly greater with SC than that with ID (P<0.05).There was no statistically significant difference between SC and ID administration of CBTX-A.Conclusions BOTOX diffuses less than CBTX-A.ID injection technique may result in less diffusion than SC.%目的 探讨两种注射用A型肉毒毒素(保妥适和衡力)在前额的弥散性,以及皮内注射和皮下注射对药物弥散性的影响.方法 选取健康志愿者20名,在前额4个部位分别注射0.05 ml(2 U)的A型肉毒毒素.随机选择在一侧前额的中部和颞侧注射保妥适,而在另一侧前额的中部和颞侧注射衡力;其中前额中部行皮下注射,颞侧行皮内注射.注射后第14天进

  6. Botulinum toxin in the management of sialorrhoea in acquired brain injury

    LENUS (Irish Health Repository)

    Carroll, A

    2016-06-01

    Sialorrhoea as a consequence of severe acquired brain injury can significantly negatively impact on quality of life. Medications used in its management have many side effects which can cause problems in the severely disabled. Botulinum toxin is an effective treatment of sialorrhoea in a number of neurological conditions but may also have a role to play in the management of sialorrhoea following severe ABI. We report on 4 cases of sialorrhoea following acquired brain injury causing a variety of problems, whose parotid glands were injected with Botulinum toxin type A (Dysport) 50mu each, under ultrasound guidance. All cases had a clinically and statistically significant reduction in drooling as measured by the teacher drooling scale (p=0.005) and carers Visual Analogue Scale (p=0.012). There were no side effects reported. Botulinum toxin is an effective treatment for sialorrhoea associated with acquired brain injury.

  7. God behandlingseffekt af botulinum toxin A hos systemisk sklerodermi patienter med digitale sår

    DEFF Research Database (Denmark)

    Nielsen, Jane Baumgartner; Hvid, Lone; Olesen, Anne Braae

    2014-01-01

    In this case we describe a successful combined treatment with local anaesthetics and botulinum toxin A. A 61-year-old man with systemic sclerosis of limited type presented treatment refractory digital ulcers on his toes with a poor response to conventional treatment. A combined treatment as above......-mentioned prevented a threatening amputation and improved quality of life, reduction of pain and healing of wounds. Using botulinum toxin A combined with local anaesthetics to severe toe digital ulcers in patients with systemic sclerosis may be a solution, when other treatments have been ineffective and amputation...

  8. Toxina botulínica tipo A tópica con iontoforesis para el tratamiento de la hiperhidrosis axilar: Efecto y persistencia Topic type A botulinum toxin with iontophoresis in the treatment of armpit hyperhidrosis: effect and persistency

    Directory of Open Access Journals (Sweden)

    J.F. Silva-Gavarrete

    2011-09-01

    exocrine glands is termed Hyperhidrosis (HH and frequently become a dermatologic and social problem for humans. Nowadays, we have multiple treatments that controls the armpit HH. Botulinum toxin type A (TXB-A is known to be the best treatment to eliminate this problem but the needing of multiple injections in the armpit limits patients´ acceptance. Clinical iontophoresis method uses galvanic current to introduce many transdermal medications. We perform a simple blind clinical assay over 10 patients with armpit HH in who we apply an inert gel blended with Botulinum toxin type A (TXB-A Dysport® using one session of iontopheresis in one armpit; in the same moment the other armpit was injected with the toxin in the conventional way. The results where evaluated and compare by Minor Test (starch-iodine test in each patient at day 10th, 2 months and 5 months after the application. The same number of units and dilution of TXB-A where used in the topic and injected administration way. Results shows a diminished armpit HH in both sides over the whole study, been higher percentage of the effect in the injected way. In general a 74.67% decrease of armpit sweat for the topical way with iontopheresis and 90.33% of decrease of armpit sweat for the injected way. In the 5 months control of the persistency of the effect, both ways of administration of TXB-A reports with statistical significant results. Therefore in the present study we conclude that TXB-A apply topically with iontopheresis improves the armpit HH and shows a persistency of the effect at least for 5 months period.

  9. Serum Antibody Response to Clostridium botulinum Toxin in Infant Botulism

    OpenAIRE

    Rubin, Lorry G.; Dezfulian, Manuchehr; Yolken, Robert H.

    1982-01-01

    A serum antibody response has not been previously demonstrated after infection with Clostridium botulinum. We developed an enzyme immunoassay for measuring serum antibody to C. botulinum toxins A, B, and E. This assay system detected a specific immunoglobulin G and immunoglobulin M antibody response to C. botulinum toxin in two patients with infant botulism.

  10. Specificity of botulinum protease for human VAMP family proteins.

    Science.gov (United States)

    Yamamoto, Hideyuki; Ida, Tomoaki; Tsutsuki, Hiroyasu; Mori, Masatoshi; Matsumoto, Tomoko; Kohda, Tomoko; Mukamoto, Masafumi; Goshima, Naoki; Kozaki, Shunji; Ihara, Hideshi

    2012-04-01

    The botulinum neurotoxin light chain (BoNT-LC) is a zinc-dependent metalloprotease that cleaves neuronal SNARE proteins such as SNAP-25, VAMP2, and Syntaxin1. This cleavage interferes with the neurotransmitter release of peripheral neurons and results in flaccid paralysis. SNAP, VAMP, and Syntaxin are representative of large families of proteins that mediate most membrane fusion reactions, as well as both neuronal and non-neuronal exocytotic events in eukaryotic cells. Neuron-specific SNARE proteins, which are target substrates of BoNT, have been well studied; however, it is unclear whether other SNARE proteins are also proteolyzed by BoNT. Herein, we define the substrate specificity of BoNT-LC/B, /D, and /F towards recombinant human VAMP family proteins. We demonstrate that LC/B, /D, and /F are able to cleave VAMP1, 2, and 3, but no other VAMP family proteins. Kinetic analysis revealed that all LC have higher affinity and catalytic activity for the non-neuronal SNARE isoform VAMP3 than for the neuronal VAMP1 and 2 isoforms. LC/D in particular exhibited extremely low catalytic activity towards VAMP1 relative to other interactions, which we determined through point mutation analysis to be a result of the Ile present at residue 48 of VAMP1. We also identified the VAMP3 cleavage sites to be at the Gln 59-Phe 60 (LC/B), Lys 42-Leu 43 (LC/D), and Gln 41-Lys 42 (LC/F) peptide bonds, which correspond to those of VAMP1 or 2. Understanding the substrate specificity and kinetic characteristics of BoNT towards human SNARE proteins may aid in the development of novel therapeutic uses for BoNT.

  11. Neurorehabilitation with versus without resistance training after botulinum toxin treatment in children with cerebral palsy

    DEFF Research Database (Denmark)

    Bandholm, Thomas Quaade; Jensen, Bente Rona; Nielsen, Lone M;

    2012-01-01

    Objective: To compare the effects of physical rehabilitation with (PRT) and without (CON) progressive resistance training following treatment of spastic plantarflexors with botulinum toxin type A (BoNT) in children with cerebral palsy (CP). Methods: Fourteen children with CP performed supervised...

  12. Botulinum therapy for poststroke spasticity of the lower extremity (clinical cases

    Directory of Open Access Journals (Sweden)

    L. V. Krylova

    2014-01-01

    Full Text Available The paper deals with the topical problem – the medical rehabilitation of patients with poststroke spasticity. It describes clinical cases of patients with poststroke spasticity of the upper and lower extremities who have received combined therapy using botulinum toxin type A (Botox injections.

  13. Thickened saliva after effective management of drooling with botulinum toxin A.

    NARCIS (Netherlands)

    Erasmus, C.E.; Hulst, K. van; Hoogen, F.J.A. van den; Limbeek, J. van; Roeleveld, N.; Veerman, E.C.; Rotteveel, J.J.; Jongerius, P.H.

    2010-01-01

    AIM: The aim of this study was to evaluate the rheological properties of saliva after submandibular botulinum toxin type A (BoNT-A) injections. METHOD: We enrolled 15 children (11 males and six females; age range 3-17 y, mean age 9 y 10 mo) diagnosed with spastic (n=9) or dyskinetic (n=6) quadripleg

  14. Thickened saliva after effective management of drooling with botulinum toxin A

    NARCIS (Netherlands)

    C.E. Erasmus; K. van Hulst; F.J. van den Hoogen; J. van Limbeek; N. Roeleveld; E.C.I. Veerman; J.J. Rotteveel; P.H. Jongerius

    2010-01-01

    Aim: The aim of this study was to evaluate the rheological properties of saliva after submandibular botulinum toxin type A (BoNT-A) injections. Method: We enrolled 15 children (11 males and six females; age range 3-17y, mean age 9y 10mo) diagnosed with spastic (n=9) or dyskinetic (n=6) quadriplegic

  15. Thickened Saliva after Effective Management of Drooling with Botulinum Toxin A

    Science.gov (United States)

    Erasmus, Corrie E.; van Hulst, Karen; van den Hoogen, Frank J. A.; van Limbeek, Jacques; Roeleveld, Nel; Veerman, Enno C. I.; Rotteveel, Jan J.; Jongerius, Peter H.

    2010-01-01

    Aim: The aim of this study was to evaluate the rheological properties of saliva after submandibular botulinum toxin type A (BoNT-A) injections. Method: We enrolled 15 children (11 males and six females; age range 3-17y, mean age 9y 10mo) diagnosed with spastic (n=9) or dyskinetic (n=6) quadriplegic cerebral palsy (CP); Gross Motor Function…

  16. Tratamiento de epicondilitis refractaria con neurotoxina botulínica tipo A libre de complejo proteínico Treament of refractory epicondilitis with botulin neurotoxin type A free of protein complex

    Directory of Open Access Journals (Sweden)

    B. Santos

    2013-02-01

    conservative measures are failing. Material and methods: We present 3 patients diagnosed of epicondylitis for more than 6 months, and which had been subjected to the usual treatment without success, which will manage with free protein complex botulinum toxin (50 IU at a single point of application on an outpatient basis. The intensity of the pain in the 3 patients were evaluated using an analog scale visual (EVA in which 0 represents no pain and 10 means more intense pain experienced with resisted wrist extension maneuver both palpation. Results: We obtained a significant reduction of pain in 3 patients treated to the first month of his administration with total resolution at 3 months. This benefit was maintained at least during 6 months. Conclusions: refractory to conservative treatment can be effectively treated with free protein complex botulinum toxin.

  17. Use of Botulinum toxin in 55 children with cerebral palsy

    Directory of Open Access Journals (Sweden)

    Mohammadi M

    2000-10-01

    Full Text Available Botulinum toxin A (BTA inhibits presynaptic release of acetylcholine at the neuromuscular junction and has reportedly been successful in the treatment of spastic disorders.To evaluate the effect of botulinum toxin on cerebral palsied children with spastic or mixed type of the disease, especially those patiens having spasticity as a cardinal symptom without joint contracture, we designed the following study. Ninety-one cases (55 of referred patients to pediatic Neurology outpatient clinics of children’s Medical Center were given BTA injections in affected muscles of the lower limb. They were reevaluating 3 to 5 weeks and 3 months later for type of walking and range of affected joints’ movement. The study showed a clinically significant gait improvement in 71.2% of patients (P<0.0005 and also an overall increased range of motion in affected limbs after BTA injection (P<0.04. Side effects occurred only in two cases as transient generalized weakness, gent recurvatum and ptosis. Drug effectiveness was time-limited, lasting abot 3 months in all patients ( a golden time for rehabilitation therapists to improve the patients’ condition. Overall, BTA has improved both the type of walking as well as the range of joints motion in our patients. So its’ administration is suggested in cerebral palsied children if the spasticity is a major and disabling sign

  18. The analgesic effects of intra-articular botulinum toxin type A and mechanisms in chronic knee arthritis pain model%A型肉毒杆菌毒素对佐剂关节炎大鼠的镇痛作用

    Institute of Scientific and Technical Information of China (English)

    茹靖涛; 曹靖; 秦涛

    2012-01-01

    目的 观察关节腔内注射A型肉毒杆菌毒素(BoNT/A)在佐剂关节炎模型中的镇痛效果及疼痛相关物质在脊神经节及脊髓的变化.方法 SD大鼠右后足跖部皮下注射完全弗氏佐剂(CFA)0.1 ml建立类风湿性关节炎的动物模型(AA).关节炎形成后随机分为3组:BoNT组(n=10):关节腔内注射Botox/A 0.02 IU/5μl;NS组(n=10):关节腔内注射生理盐水5μl;Sham组(n=10):非关节炎模型组.5d后检测疼痛行为学、力量评估;免疫组织化学检测脊神经节钠离子通道1.8(Nav1.8)、脊髓N-甲基-D-天冬氨酸受体2B亚基(NR2B)的表达.结果 肉眼步态分析显示大鼠关节炎引起的严重的步态异常,BoNT治疗组改善了40% (P <0.05);治疗组大鼠右足热痛阈升高(P<0.05);脊神经节Nav1.8高表达;脊髓NR2B表达降低.结论 通过关节腔内注射Botox/A可以抑制疼痛信号的传导,减轻关节痛.%Objective To estimate the analgesic effect of intra-articular injection of botulinum toxin type A (BoNT/A) in adjuvant arthritis model and to explore the related factors associated with arthritis pain.Methods We established murime model of chronic knee arthritis pain by injection of complete Freund's adjuvant (CFA) 0.1 ml into the right paw of Sprague Dawley rats.Following induction of arthritis,BoNT/A group (n =10) received intra-articular BoNT/A0.02 IU/5 μL; NS group (n =10) was treated with intra-articular 5 μL saline injections.In Sham group (n =10),no artiritis model was established.Pain behavior testing was performed prior to arthritis,after induction of arthritis,and following treatments.Pain behavior measures included analysis of gait impairment and thermal pain threshold.Strength was measured as ability to cling.Pain related factors Nav1.8 and NR2B were detected by using immunohistochemistry.Results Gait analysis showed that significant impairment of gait in arthritic mice wast improved by 40% ( P < 0.05 ) after intra-articular Bo

  19. Treatment of scar contracture with intralesional botulinum toxin type A : a preliminary research%A型肉毒毒素治疗挛缩性瘢痕的初步研究

    Institute of Scientific and Technical Information of China (English)

    颜彤彤; 陈敏亮; 马奎; 孙同柱; 梁黎明; 刘畅; 赖林英; 付小兵

    2013-01-01

    Objective: To evaluate the effect and mechanism of botulinum toxin type A (BTXA) in treating scar contracture. Methods: Ten patients with typical contractive scars were involved in this study. Two independent scars (area <10 cm2) of each patient were selected and were randomly divided into BTXA group (receiving intralesional injection of BTXA for 2 times trimonthly) , and blank control group (receiving the same volume of normal saline). Before and 1,3,6 months after BTXA injection, the length of the longest axis of the scar was measured to reflect the changes in scar contracture. The scar tissues from 5 patients undergoing excision of scar 6 months after BTXA injection were observed with HE staining by light microscopy, and the expression of α-smooth muscle actin (α-SMA) and myosin-Ⅱ were assayed by immunohistochemical method. Results: The degree of scar contraction was lessened in BTXA group compared with that of control group (P<0. 01) , especially 6 months after the first injection [(1. 103 ± 0. 158)cm vs. (0. 187 ± 0. 221)cm]. The collagen was sparse and better aligned, and the number of fibroblasts was significantly reduced at the BTXA injection area. The expression of α-SMA and myosin-Ⅱ also decreased in BTXA group(P<0. 01). Conclusion: The results show that BTXA injections is safe and effective in treating contractive scars. The mechanism maybe attributable to a reduction of the expression of a-SMA and myosin- Ⅱ of fibroblasts in scars.%目的:探讨A型肉毒毒素(BTXA)治疗挛缩性瘢痕的效果及其作用机制.方法:10例烧伤后挛缩性瘢痕患者,每人选取两处挛缩性瘢痕,随机分为BTXA组(瘢痕内注射BTXA,间隔3个月1次,共2次)和空白对照组(瘢痕内注射等积生理盐水),于注射前及首次注射后1、3、6个月测量瘢痕长轴长度,观察其长度变化用以反映瘢痕挛缩程度;切取5例药物治疗6个月后行瘢痕切除手术患者的瘢痕组织,HE染色观察病理学改变,免疫组

  20. Effect of Botulinum Toxin Type A and Jin Three-needle on Treatment of Tip Toes of Children Patient with Spastic Diplegia Cerebral Palsy%A型肉毒毒素与靳三针治疗痉挛型双瘫脑瘫患儿尖足步态的疗效观察

    Institute of Scientific and Technical Information of China (English)

    李金玲; 徐开寿; 李翠玲; 潭红香; 靳晓坤; 郑玉蔼; 麦坚凝

    2012-01-01

    目的 观察A型肉毒毒素注射结合靳三针疗法纠正痉挛型双瘫脑瘫患儿尖足步态的疗效.方法 78例以尖足步态为主的痉挛型双瘫脑瘫患儿,均采用A型肉毒毒素局部注射治疗后,随机分为治疗组40例和对照组38例,两组患儿均采用综合康复治疗的方法,治疗组在此基础上增加靳三针疗法.在治疗前、治疗2周、3个月和6个月时对肌张力、运动功能进行评定.结果 治疗后,两组患儿的观测指标均有改善,治疗组优于对照组(P<0.05).结论 A型肉毒毒素注射结合靳三针疗法治疗痉挛型双瘫脑瘫患儿疗效显著,靳三针疗法可以提高康复治疗的效果.%Objective To observe treatment effect of Jin three-needle combined with injection of Botulinum toxin type A for children with spastic diplegia cerebral palsy. Methods 78 children patients with spastic diplegia were received local injection of botulinum toxin A, and randomly divided into treatment group (40 cases) and control group (38 cases). The two groups were carried out comprehensive rehabilitation therapy. The treatment group was received traditional medicine Jin three-needle therapy. The muscle tension and motor function of patients were evaluated before and after 2 weeks, 3 months, and 6 months treatment. Results The muscle tension, motor function and other index in two groups after treatment were improved, the treatment group were much better than that of control group (P < 0.05 ). Conclusion The combined injection of Botulinum toxin type A with Jin three-needle have significant effect on treatment of children patients with spastic diplegia cerebral palsy; and Jin three-needle could improve the curative effect of rehabilitation.

  1. Clinical research on botulinum toxin type A combining rehabilitation training for the treatment of children cerebral palsy%肉毒杆菌毒素A辅助复健训练治疗儿童痉挛型脑瘫的疗效观察

    Institute of Scientific and Technical Information of China (English)

    吴素英

    2011-01-01

    AIM: To observe the clinical effect of botulinum toxin type A combining rehabilita-tion for the treatment of children cerebral palsy METHODS: Thirty-two cases of children cere-bral palsy admitted to hospital from March 2009 to March 2011 were randomly divided into two groups, control group (n= 16) and treatment group (n= 16). The control group was given conventional rehabilitation training; the treatment group was also given injection of botulinum toxin A. The clinical effects in 2 groups were observed and compared. RESULTS: After treatment, the GMFM score in the treatment group was significantly higher than that in the control group (P<0. 05). The Ashworth score in the treatment group was significantly lower thanthat in the control group (P<0. 05); and the popliteal fossa angle, foot disaffection angle and abduction angle in the treatment group were superior to those in the control group (P<0. 05). CONCLUSION- There is a significant clinical effect of botulinum toxin type A combining rehabilitation for the treatment of children cerebral palsy.%目的:探讨采用肉毒杆菌毒素A辅助复健训练治疗儿童痉挛型脑瘫的临床疗效.方法:将本院2009年3月-2011年3月收治的32例脑性瘫痪患儿随机分成2组:对照组16例,试验组16例.其中,对照组给予常规的复健训练治疗,试验组则给予注射肉毒杆菌毒素A辅助复健训练治疗.结果:经过治疗后,试验组的粗大运动(GMFM)功能评分显著高于对照组(P<0.05);试验组的肌张力改良Ashworth评分相比对照组显著性降低(P<0.05);试验组的腘窝角、足背屈角、足外展角的活动范围相比对照组均有显著性改善(P<0.05).结论:采用肉毒杆菌毒素A辅助复健训练治疗儿童脑瘫的临床疗效显著.

  2. A型肉毒毒素联合Lux1540-非剥脱点阵激光治疗额部皱纹的临床疗效和安全性观察%CLINICAL EFFECT AND SAFETY OBSERBATION OF BOTULINUM TOXIN TYPE A INJECTION COMBINED WITH LUX 1540 FRACTIONAL LASER ON WRINKLES

    Institute of Scientific and Technical Information of China (English)

    苏雪莲

    2015-01-01

    Objective To evaluate the effectiveness and safety of botulinum toxin type A injection andLux1540fractional laser on treatment of eye wrinkles.Methods Sixty patients with wrinkles,aged from 27 to65,were divided into three groups randomly.They were treated with botulinum toxin type A,Lux1540 fractional laser,or the combined therapy.Pictures were taken for each patient before treatment and immediate,at 7 daysand 1,3,6months after treatment,with which the results were evaluated.ResultsAt 3 to 7days after treatment,the dynamic wrinkles of patients treated with botnlinum toxin type A injection were alleviated. The static wrinkles of patients treated with Lux1540 fractional laser with or without botulinum toxin type A injection were alleviated right after the treatment.The dynamic wrinkles of those who treated with the combined therapy got better long—term effect.Conclusion It is effective and safe to treat eye wrinkles by using botulinum toxin type A injection and Lux1540 fractional laser combined therapy.%目的:探讨A型肉毒毒素联合Lux1540-非剥脱点阵激光:点阵激光治疗额头皱纹的临床疗效和安全性。方法将年龄27~65岁且寻求改善额头皱纹的门诊患者60例,随机平均分为3组,分别给予A型肉毒毒素注射,Lux1540-非剥脱点阵激光治疗,A型肉毒毒素注射联合Lux1540-非剥脱点阵激光治疗。每例患者在治疗前,治疗后即刻,治疗后7 d、1、3、6个月进行随访评价。结果单纯接受肉毒毒素注射的患者,在治疗后3~7 d开始出现动态皱纹的改善;接受Lux1540-非剥脱点阵激光以及A型肉毒毒素注射联合Lux1540-非剥脱点阵激光治疗的患者,术后即刻可观察到静态皱纹的改善,且肉毒毒素注射联合Lux1540-非剥脱点阵激光的患者,其动态皱纹的远期效果改善更加明显。结论 A型肉毒毒素联合Lux1540-非剥脱点阵激光,治疗可同时改善额部的动态皱纹和静态皱纹,并有

  3. Neurotoxins from Marine Dinoflagellates: A Brief Review

    Directory of Open Access Journals (Sweden)

    Da-Zhi Wang

    2008-06-01

    Full Text Available Dinoflagellates are not only important marine primary producers and grazers, but also the major causative agents of harmful algal blooms. It has been reported that many dinoflagellate species can produce various natural toxins. These toxins can be extremely toxic and many of them are effective at far lower dosages than conventional chemical agents. Consumption of seafood contaminated by algal toxins results in various seafood poisoning syndromes: paralytic shellfish poisoning (PSP, neurotoxic shellfish poisoning (NSP, amnesic shellfish poisoning (ASP, diarrheic shellfish poisoning (DSP, ciguatera fish poisoning (CFP and azaspiracid shellfish poisoning (ASP. Most of these poisonings are caused by neurotoxins which present themselves with highly specific effects on the nervous system of animals, including humans, by interfering with nerve impulse transmission. Neurotoxins are a varied group of compounds, both chemically and pharmacologically. They vary in both chemical structure and mechanism of action, and produce very distinct biological effects, which provides a potential application of these toxins in pharmacology and toxicology. This review summarizes the origin, structure and clinical symptoms of PSP, NSP, CFP, AZP, yessotoxin and palytoxin produced by marine dinoflagellates, as well as their molecular mechanisms of action on voltage-gated ion channels.

  4. Occurrence of C. botulinum in healthy cattle and their environment following poultry botulism outbreaks in mixed farms.

    Science.gov (United States)

    Souillard, R; Le Maréchal, C; Hollebecque, F; Rouxel, S; Barbé, A; Houard, E; Léon, D; Poëzévara, T; Fach, P; Woudstra, C; Mahé, F; Chemaly, M; Le Bouquin, S

    2015-10-22

    Ten cattle farms located in an area with a recent history of poultry botulism outbreaks were investigated to evaluate the occurrence of toxigenic C. botulinum in healthy cattle. Environmental samples in the 10 cattle farms and bovine fecal contents in farms with a confirmed environmental contamination were collected. Detection of C. botulinum toxin genes C, D, C/D, D/C and E was performed using real-time PCR. 4.9% (7/143) of the environmental samples collected in the 10 investigated cattle farms were positive for C. botulinum type C/D. Theses samples (boot-swabs in stalls and on pasture and water of a stream) were collected in 3 different farms. One cow dung sample and 3 out of 64 fecal contents samples collected in a single farm were also positive for C. botulinum type C/D. This study demonstrates that cattle are probably indirectly contaminated via poultry botulism in the area and that they can be intermittent carrier of C. botulinum type C/D after poultry botulism outbreaks in mixed farms.

  5. Study on potential Clostridium botulinum growth and toxin production in Parma ham

    Directory of Open Access Journals (Sweden)

    Giuseppe Merialdi

    2016-04-01

    Full Text Available The objective of this study was to investigate Clostridium botulinum growth and toxin production in the industrially manufactured Italian Parma ham. The study focuses on the Parma ham production phase identified as maximum risk to C. botulinum proliferation, i.e. the transition from cold phase (salting and resting to a phase carried out at temperature between 15 and 23°C (drying. A preliminary in vitro test was carried out in order to verify the capability of 6 C. botulinum strains (1 type A, 4 type B, and 1 type E strains to grow in conditions of temperature, pH and NaCl concentration comparable to those of the beginning stage of ham drying. Five C. botulinum strains grew at 20°C and pH 6, four strains produced toxin when inoculated at a concentration equal to 103 cfu/mL at NaCl concentration of 4%, while when the inoculum concentration was 10 cfu/mL, NaCl concentration of 3% resulted the toxin-genesis limiting factor. An experimental contamination with a mixture of the 5 C. botulinum strains selected by the preliminary in vitro test was performed on 9 thighs inoculated at the end of the resting phase. The study was designed to evaluate the potential growth and toxin production in extremely favourable conditions for the bacterium. Type B proteolytic C. botulinum toxin was produced after 14 days of incubation at 20°C in 2 thighs characterised by high weight, low number of days of resting and anomalous physiochemical characteristics [one for very low NaCl concentration (1.59%, the other for elevated pH (6.27 and both for high water activity values (>0.970]. The results of this research confirm that the cold resting step is a critical phase in the production process of Parma ham for the investigated hazard. Based on the present study, the long resting phase adopted in the manufacturing of Parma ham is proven effective to prevent the growth of C. botulinum, an event which could not otherwise be excluded if the hams were processed under less

  6. 外周神经电刺激引导下A型肉毒毒素注射在痉挛型脑性瘫痪中的应用%Injection of Botulinum Toxin Type A Guided by Peripheral Nerve Stimulation for Spastic Cerebral Palsy

    Institute of Scientific and Technical Information of China (English)

    邵银进; 吴桂华; 曾康华

    2013-01-01

    Objective To investigate the effects of local injection of Botulinum toxin type A (BTX-A) on spastic cerebral palsy guided by peripheral nerve stimulation. Methods 30 children with spastic cerebral palsy received local multi-point injection of BTX-A guided by peripheral nerve stimulation. They were assessed with modified Ashworth scale (MAS) and Gross Motor Function Assessment Scale (GM-FM-88) before and 1 week, 3 months and 6 months after treatment. Results The scores of MAS decreased significantly 1 week, 3 months and 6 months after treatment (P0.05),3个月和6个月时显著增加(P<0.001)。结论外周神经电刺激引导下BTX-A局部多点注射治疗痉挛型脑瘫疗效显著。

  7. Toxina botulínica tipo A para el manejo del dolor en pacientes con síndrome de dolor miofascial crónico Botulinum toxin type A for the management of pain in patients with chronic myofascial pain

    Directory of Open Access Journals (Sweden)

    J.C. Torres Huerta

    2010-02-01

    Full Text Available Introducción: El síndrome de dolor miofascial es uno de los principales problemas de dolor crónico en la práctica clínica, comúnmente asociado a un traumatismo o a microtraumatismos repetitivos. La toxina botulínica tipo A (Botox® es una alternativa aceptada como opción terapéutica. Objetivo: Valorar la funcionalidad de las actividades diarias mejorando la intensidad del dolor con el uso de la toxina botulínica tipo A, infiltrada en puntos gatillo, en pacientes con síndrome de dolor miofascial crónico. Material y métodos: 30 pacientes con síndrome de dolor miofascial crónico con edad entre 25 y 50 años de ambos sexos. A todos se les realizó infiltración de puntos gatillo mediante equipo de electrosonomiografía para la aplicación de toxina botulínica tipo A a una dilución de las 100 Um de toxina en 1 ml de solución salina infiltrando de 200 a 400 Um totales, dependiendo del sitio del dolor. Se valoró la calidad analgésica mediante la escala visual analógica (EVA y el índice de funcionalidad con el cuestionario de Oswestry durante 4 semanas. Resultados: La valoración de la EVA promedio inicial fue de 7,23 y la final de 3,13 (p Introduction: Myofascial syndrome is one of the main chronic pain syndromes encountered in clinical practice and is commonly associated with trauma or repetitive microtrauma. Botulinum toxin type A (Botox® is an accepted therapeutic option. Objective: To evaluate the use of botulinum toxin type A (Botox®, infiltrated in trigger points, as an option in the management of pain in patients with chronic myofascial syndrome. Material and Methods: Thirty men and women with myofascial pain syndrome, aged between 25 and 50 years were included. In all patients, botulinum toxin type A (Botox® was infiltrated in trigger points by means of electrosonomyographic equipment. A dilution of 100 Um toxin in 1 cc saline solution was used and a total of 200 to 400 Um was administered depending on the site of the

  8. The change of substance P in neuropathic pain rats injected subcutaneous-ly by botulinum toxin type A%A型肉毒毒素对神经病理性疼痛模型大鼠P物质含量的影响

    Institute of Scientific and Technical Information of China (English)

    张晓英; 杨国栋; 潘万龙

    2015-01-01

    目的::研究A型肉毒毒素(botulinum toxin type A,BTX-A)对神经病理性疼痛模型大鼠P物质(substance P,SP)含量的影响。方法:5%福尔马林50μL皮下注射于大鼠额区及颞区,建立疼痛模型。 BTX-A组局部皮下注射10 U/kg BTX-A,同时设立生理盐水组及空白对照组,放射免疫法测定SP 含量。结果:生理盐水组SP 含量高于空白对照组( P<0.05), BTX-A组低于生理盐水组(P<0.05)。结论: BTX-A可能通过抑制感觉通路神经肽-SP的释放减轻神经病理性疼痛症状。%Objective:To detect the change of substance P in neuropathic pain rats injected subcutaneously by botulinum toxin type A. Methods:Besides control group,neuropathic pain rats triggered by 5 % formalin subcutaneously injected into frontal and tem-poral areas,then they had been respectively treated with 0. 9 % NaCl (n=10)and 10 U/kg BTX-A (n=10) for 6 days. All of groups were detected the content of substance P(SP) by radioimmunoassay. Results:The content of SP increased significantly in saline group than that in control group (P<0. 05),but decreased significantly in BTX-A groups than in saline group(P<0. 05). Conclusion:BTX-A could inhibit SP releasing from sensory pathways to relieve the symptoms of neuropathic pain.

  9. Avaliação do filme lacrimal de pacientes com distonia facial durante tratamento com toxina botulínica tipo A Lacrimal film evaluation of patients with facial dystonia during botulinum toxin type A treatment

    Directory of Open Access Journals (Sweden)

    Patricia Grativol Costa

    2006-06-01

    Full Text Available OBJETIVO: Determinar o efeito da toxina botulínica no filme lacrimal em pacientes com distonia facial. MÉTODOS: Foram incluídos 24 pacientes portadores de blefaroespasmo essencial e espasmo hemifacial que receberam aplicação de toxina botulínica tipo A que foram submetidos à propedêutica do filme lacrimal previamente à aplicação e após, com 7 e 30 dias. RESULTADOS: Houve diminuição das queixas de olho seco trinta dias após a aplicação, entretanto, o tempo de ruptura do filme lacrimal e o teste de Schirmer não demonstraram variação significativa entre os períodos pré-tratamento e 1 mês da aplicação. Em relação ao teste de coloração com rosa bengala, todos os olhos que coraram no pré-tratamento, melhoraram na última avaliação. CONCLUSÃO: A injeção de toxina botulínica pode aliviar as queixas de olho seco nos pacientes com distonia facial pela provável ação de inibição do orbicular na sua função de bomba lacrimal.PURPOSE: To determine the effect of botulinum toxin injection in the eyelid on lacrimal film in patients with facial dystonia. METHODS: Twenty-four patients with essential blepharospasm and hemifacial spasm were submitted to botulinum toxin injection and lacrimal film tests were performed before the application and after seven and thirty days. RESULTS: There was improvement in symptoms of dry eye and rose bengal test, however, the breakup time and Schirmer's test did not show significant variation between pretreatment and after 1 month of follow-up. CONCLUSION: The dry eye symptoms in patients with facial dystonia may be attenuated by botulinum toxin due to its possible inhibitory effect on the orbicular muscle leading to a decrease in lacrimal pump.

  10. Treatment of severe drooling with botulinum toxin in amyotrophic lateral sclerosis and Parkinson's disease: Efficacy and possible mechanisms

    DEFF Research Database (Denmark)

    Møller, Eigild; Karlsborg, Merete; Jensen, Allan Bardow;

    2011-01-01

    Drooling in neurodegenerative diseases is associated with social impediment. Previous treatments of drooling have little effect or are effective but with severe side effects. Therefore, there is a need to test new methods such as the use of botulinum toxin type A (BTX-A).......Drooling in neurodegenerative diseases is associated with social impediment. Previous treatments of drooling have little effect or are effective but with severe side effects. Therefore, there is a need to test new methods such as the use of botulinum toxin type A (BTX-A)....

  11. New Insights into the genetic diversity of Clostridium botulinum Group III through extensive genome exploration

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    Cédric eWoudstra

    2016-05-01

    Full Text Available Animal botulism is caused by group III Clostridium botulinum strains producing type C and D toxins, or their chimeric forms C/D and D/C. Animal botulism is considered an emerging disease in Europe, notably in poultry production. Before our study, 14 genomes from different countries were available in the public database, but none were from France. In order to investigate the genetic relationship of French strains with different geographical areas and find new potential typing targets, 17 strains of C. botulinum group III were sequenced (16 from France and one from New Caledonia. Fourteen were type C/D strains isolated from chickens, ducks, guinea fowl and turkeys and three were type D/C strains isolated from cattle. The New Caledonian strain was a type D/C strain. Whole genome sequence analysis showed the French strains to be closely related to European strains from C. botulinum group III lineages Ia and Ib. The investigation of CRISPR sequences as genetic targets for differentiating strains in group III proved to be irrelevant for type C/D due to a deficient CRISPR/Cas mechanism, but not for type D/C. Conversely, the extrachromosomal elements of type C/D strains could be used to generate a genetic ID card. The highest level of discrimination was achieved with SNP core phylogeny, which allowed differentiation up to strain level and provide the most relevant information for genetic epidemiology studies and discrimination.

  12. New Insights into the Genetic Diversity of Clostridium botulinum Group III through Extensive Genome Exploration.

    Science.gov (United States)

    Woudstra, Cédric; Le Maréchal, Caroline; Souillard, Rozenn; Bayon-Auboyer, Marie-Hélène; Mermoud, Isabelle; Desoutter, Denise; Fach, Patrick

    2016-01-01

    Animal botulism is caused by group III Clostridium botulinum strains producing type C and D toxins, or their chimeric forms C/D and D/C. Animal botulism is considered an emerging disease in Europe, notably in poultry production. Before our study, 14 genomes from different countries were available in the public database, but none were from France. In order to investigate the genetic relationship of French strains with different geographical areas and find new potential typing targets, 17 strains of C. botulinum group III were sequenced (16 from France and one from New Caledonia). Fourteen were type C/D strains isolated from chickens, ducks, guinea fowl and turkeys and three were type D/C strains isolated from cattle. The New Caledonian strain was a type D/C strain. Whole genome sequence analysis showed the French strains to be closely related to European strains from C. botulinum group III lineages Ia and Ib. The investigation of CRISPR sequences as genetic targets for differentiating strains in group III proved to be irrelevant for type C/D due to a deficient CRISPR/Cas mechanism, but not for type D/C. Conversely, the extrachromosomal elements of type C/D strains could be used to generate a genetic ID card. The highest level of discrimination was achieved with SNP core phylogeny, which allowed differentiation up to strain level and provide the most relevant information for genetic epidemiology studies and discrimination.

  13. Identification of novel linear megaplasmids carrying a ß-lactamase gene in neurotoxigenic Clostridium butyricum type E strains.

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    Giovanna Franciosa

    Full Text Available Since the first isolation of type E botulinum toxin-producing Clostridium butyricum from two infant botulism cases in Italy in 1984, this peculiar microorganism has been implicated in different forms of botulism worldwide. By applying particular pulsed-field gel electrophoresis run conditions, we were able to show for the first time that ten neurotoxigenic C. butyricum type E strains originated from Italy and China have linear megaplasmids in their genomes. At least four different megaplasmid sizes were identified among the ten neurotoxigenic C. butyricum type E strains. Each isolate displayed a single sized megaplasmid that was shown to possess a linear structure by ATP-dependent exonuclease digestion. Some of the neurotoxigenic C. butyricum type E strains possessed additional smaller circular plasmids. In order to investigate the genetic content of the newly identified megaplasmids, selected gene probes were designed and used in Southern hybridization experiments. Our results revealed that the type E botulinum neurotoxin gene was chromosome-located in all neurotoxigenic C. butyricum type E strains. Similar results were obtained with the 16S rRNA, the tetracycline tet(P and the lincomycin resistance protein lmrB gene probes. A specific mobA gene probe only hybridized to the smaller plasmids of the Italian C. butyricum type E strains. Of note, a ß-lactamase gene probe hybridized to the megaplasmids of eight neurotoxigenic C. butyricum type E strains, of which seven from clinical sources and the remaining one from a food implicated in foodborne botulism, whereas this ß-lactam antibiotic resistance gene was absent form the megaplasmids of the two soil strains examined. The widespread occurrence among C. butyricum type E strains associated to human disease of linear megaplasmids harboring an antibiotic resistance gene strongly suggests that the megaplasmids could have played an important role in the emergence of C. butyricum type E as a human

  14. Treatment of Frontal Hyperhidrosis With Botulinum Toxin

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    Ayşe Esra Koku Aksu

    Full Text Available Focal hyperhidrosis is usually localized to the axillae, palms and soles. Less frequently, hyperhidrosis may be confined to the forehead and may have negative impact on patient’s quality of life. A 34-year-old man presented to our clinic with the complaint of frontal hyperhidrosis. He was treated with botulinum toxin A. Thirty points were marked over the forehead and at each injection point, 0.15 ml (3U botulinum toxin A were injected intracutaneously. Hyperhidrosis was significantly reduced and the effect lasted for 12 months. Skindex-29, a quality-of-life measure for skin disease, was administered to the patient at the beginning and at the end of second week of botulinum toxin A injection. There was a significant improvement on the Skindex-29 scale at the end of the treatment. There was no any side effect detected during and after the treatment. Botulinum toxin A treatment is considered to be effective and safe for frontal hyperhidrosis.

  15. A unique restriction site in the flaA gene allows rapid differentiation of group I and group II Clostridium botulinum strains by PCR-restriction fragment length polymorphism analysis.

    Science.gov (United States)

    Paul, Catherine J; Tran, Shulin; Tam, Kevin J; Austin, John W

    2007-09-01

    Clostridium botulinum produces the potent botulinum neurotoxin, the causative agent of botulism. Based on distinctive physiological traits, strains of C. botulinum can be divided into four groups: however, only groups I and II are associated with human illness. Alignment of the flaA gene sequences from 40 group I and 40 group II strains identified a single BsrG1 restriction cut site that was present at base pair 283 in all group II flaA sequences and was not found in any group I sequence. The flaA gene was amplified by rapid colony PCR from 22 group I strains and 18 group II strains and digested with BsrGI restriction enzyme. Standard agarose gel electrophoresis with ethidium bromide staining showed two fragments, following restriction digestion of group II flaA gene amplicons with BsrGI, but only a single band of uncut flaA from group I strains. Combining rapid colony PCR with BsrGI restriction digest of the flaA gene at 60 degrees C is a significant improvement over current methods, such as meat digestion or amplified fragment length polymorphism, as a strain can be identified as either group I or group II in under 5 h when starting with a visible plated C. botulinum colony.

  16. An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions

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    Gary S. Sayler

    2013-03-01

    Full Text Available Marine neurotoxins are natural products produced by phytoplankton and select species of invertebrates and fish. These compounds interact with voltage-gated sodium, potassium and calcium channels and modulate the flux of these ions into various cell types. This review provides a summary of marine neurotoxins, including their structures, molecular targets and pharmacologies. Saxitoxin and its derivatives, collectively referred to as paralytic shellfish toxins (PSTs, are unique among neurotoxins in that they are found in both marine and freshwater environments by organisms inhabiting two kingdoms of life. Prokaryotic cyanobacteria are responsible for PST production in freshwater systems, while eukaryotic dinoflagellates are the main producers in marine waters. Bioaccumulation by filter-feeding bivalves and fish and subsequent transfer through the food web results in the potentially fatal human illnesses, paralytic shellfish poisoning and saxitoxin pufferfish poisoning. These illnesses are a result of saxitoxin’s ability to bind to the voltage-gated sodium channel, blocking the passage of nerve impulses and leading to death via respiratory paralysis. Recent advances in saxitoxin research are discussed, including the molecular biology of toxin synthesis, new protein targets, association with metal-binding motifs and methods of detection. The eco-evolutionary role(s PSTs may serve for phytoplankton species that produce them are also discussed.

  17. Neurotoxins and their binding areas on voltage-gated sodium channels

    Directory of Open Access Journals (Sweden)

    Marijke eStevens

    2011-11-01

    Full Text Available Voltage-gated Sodium Channels (VGSCs are large transmembrane proteins that conduct sodium ions across the membrane and by doing so they generate signals of communication between many kinds of tissues. They are responsible for the generation and propagation of action potentials in excitable cells, in close collaboration with other channels like potassium channels. Genetic defects in sodium channel genes therefore can cause a wide variety of diseases, generally called ‘channelopathies’.The first insights into the mechanism of action potentials and the involvement of sodium channels originated from Hodgkin and Huxley for which they were awarded the Nobel Prize in 1963. Until now, these concepts still form the basis for understanding the functioning of VGSCs. When VGSCs sense a sufficient change in membrane potential, they are activated and will generate a massive influx of sodium ions. Immediately after, channels will start inactivating and currents decrease. In the inactivated state channels stay refractory for any new stimulus and they must return to the closed state before being susceptible to any new depolarization. On the other hand, studies with neurotoxins like tetrodotoxin (TTX and saxitoxin (STX also contributed largely to our today’s understanding of the structure and function of ion channels and specifically of VGSCs. Moreover, neurotoxins acting on ion channels turned out to be valuable tools in the development of new drugs for the enormous range of diseases in which ion channels are involved. A recent example of a synthetic neurotoxin that made it to the market is ziconotide (Prialt®, Elan. The original peptide, -MVIIA, is derived from the cone snail Conus magus and now FDA/EMEA-approved for the management of severe chronic pain by blocking the N-type voltage-gated calcium channels in neurons.This review focuses on the current status of research on neurotoxins acting on VGSC, their contribution to further unravel the

  18. Neurotoxins and their binding areas on voltage-gated sodium channels.

    Science.gov (United States)

    Stevens, Marijke; Peigneur, Steve; Tytgat, Jan

    2011-01-01

    Voltage-gated sodium channels (VGSCs) are large transmembrane proteins that conduct sodium ions across the membrane and by doing so they generate signals of communication between many kinds of tissues. They are responsible for the generation and propagation of action potentials in excitable cells, in close collaboration with other channels like potassium channels. Therefore, genetic defects in sodium channel genes can cause a wide variety of diseases, generally called "channelopathies." The first insights into the mechanism of action potentials and the involvement of sodium channels originated from Hodgkin and Huxley for which they were awarded the Nobel Prize in 1963. These concepts still form the basis for understanding the function of VGSCs. When VGSCs sense a sufficient change in membrane potential, they are activated and consequently generate a massive influx of sodium ions. Immediately after, channels will start to inactivate and currents decrease. In the inactivated state, channels stay refractory for new stimuli and they must return to the closed state before being susceptible to a new depolarization. On the other hand, studies with neurotoxins like tetrodotoxin (TTX) and saxitoxin (STX) also contributed largely to our today's understanding of the structure and function of ion channels and of VGSCs specifically. Moreover, neurotoxins acting on ion channels turned out to be valuable lead compounds in the development of new drugs for the enormous range of diseases in which ion channels are involved. A recent example of a synthetic neurotoxin that made it to the market is ziconotide (Prialt(®), Elan). The original peptide, ω-MVIIA, is derived from the cone snail Conus magus and now FDA/EMA-approved for the management of severe chronic pain by blocking the N-type voltage-gated calcium channels in pain fibers. This review focuses on the current status of research on neurotoxins acting on VGSC, their contribution to further unravel the structure and function of

  19. Plasmidome interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum converts strains of independent lineages into distinctly different pathogens.

    Directory of Open Access Journals (Sweden)

    Hanna Skarin

    Full Text Available Clostridium botulinum (group III, Clostridium novyi and Clostridium haemolyticum are well-known pathogens causing animal botulism, gas gangrene/black disease, and bacillary hemoglobinuria, respectively. A close genetic relationship exists between the species, which has resulted in the collective term C. novyi sensu lato. The pathogenic traits in these species, e.g., the botulinum neurotoxin and the novyi alpha toxin, are mainly linked to a large plasmidome consisting of plasmids and circular prophages. The plasmidome of C. novyi sensu lato has so far been poorly characterized. In this study we explored the genomic relationship of a wide range of strains of C. novyi sensu lato with a special focus on the dynamics of the plasmidome. Twenty-four genomes were sequenced from strains selected to represent as much as possible the genetic diversity in C. novyi sensu lato. Sixty-one plasmids were identified in these genomes and 28 of them were completed. The genomic comparisons revealed four separate lineages, which did not strictly correlate with the species designations. The plasmids were categorized into 13 different plasmid groups on the basis of their similarity and conservation of plasmid replication or partitioning genes. The plasmid groups, lineages and species were to a large extent entwined because plasmids and toxin genes had moved across the lineage boundaries. This dynamic process appears to be primarily driven by phages. We here present a comprehensive characterization of the complex species group C. novyi sensu lato, explaining the intermixed genetic properties. This study also provides examples how the reorganization of the botulinum toxin and the novyi alpha toxin genes within the plasmidome has affected the pathogenesis of the strains.

  20. High-dose botulinum toxin type A local injection therapy for axillary hyperhidrosis%大剂量A型肉毒毒素局部注射治疗腋部多汗症

    Institute of Scientific and Technical Information of China (English)

    高扬; 宋军; 李虎; 鲁元刚

    2011-01-01

    目的 探讨大剂量肉毒毒素治疗腋部多汗症的长期疗效和重复治疗的疗效.方法 92例患者随机分为两组:小剂量组为每侧腋部皮内注射生理盐水稀释的A型肉毒毒素50U;大剂量组为每侧腋部皮内注射生理盐水稀释的A型肉毒毒素200U;随访3~29个月,观察两组并发症,并建立两组等级资料,经χ2 检验,评价两组患者疗效差异.结果 两组疗效进行对照分析,经过统计学处理分析,认为对于腋部多汗症的患者,小剂量与大剂量的BTXA治疗方法的疗效间隔时间,差异有统计学意义.结论 大剂量A型肉毒毒素能够显著延长腋部多汗症复发间隔时间.%Objective Evaluate the long-term effectiveness of high-dose botulinum toxin therapy in axillary hyperhidrosis, the response to repeated treatment, and the possible side effects. Methods Totally 92 patients with axillary hyperhidrosis were randomly divided into two groups. One group were injected with low-dose botulinum toxin A( BTX-A ), 50 U was injected per axilla. Another group were injected with high-dose BTX-A. A total dose of 200 U of BTX-A was used per axilla. Patients were followed up for periods up to 29 months. To investigate the effect of two methods, we analyzed two ranked data by rank sum test and x2 test to judge the disparities of the therapeutic effect. Results The results showed that the relapse-free interval of two groups with axillary hyperhidrosis was significant difference through the statistical analysis. Conclusion High-dose BTX-A treatment is capable of prolonging the antihidrotic effect of intracutaneous.

  1. The use of botulinum toxin in head and face medicine: An interdisciplinary field

    Directory of Open Access Journals (Sweden)

    Laskawi Rainer

    2008-03-01

    Full Text Available Abstract Background In this review article different interdisciplinary relevant applications of botulinum toxin type A (BTA in the head and face region are demonstrated. Patients with head and face disorders of different etiology often suffer from disorders concerning their musculature (example: synkinesis in mimic muscles or gland-secretion. This leads to many problems and reduces their quality of life. The application of BTA can improve movement disorders like blepharospasm, hemifacial spasm, synkinesis following defective healing of the facial nerve, palatal tremor, severe bruxism, oromandibular dystonias hypertrophy of the masseter muscle and disorders of the autonomous nerve system like hypersalivation, hyperlacrimation, pathological sweating and intrinsic rhinitis. Conclusion The application of botulinum toxin type A is a helpful and minimally invasive treatment option to improve the quality of life in patients with head and face disorders of different quality and etiology. Side effects are rare.

  2. Clinically and electrophysiologically diagnosed botulinum intoxication.

    Science.gov (United States)

    Kotan, Dilcan; Aygul, Recep; Ceylan, Mustafa; Yilikoglu, Yalcin

    2013-01-03

    In this case report, clinical and electrophysiological findings of 43-year-old female patient who developed Clostridium botulinum intoxication after consumption of home-made canned food are presented. Following the sudden onset of severe nausea and vomiting, diplopia, blurred vision, bilateral ptosis, weakness, speech and swallowing difficulties have developed and the patient declared that she has just tasted the canned beans after she had rinsed them several times. The case, where serological tests cannot be performed, was diagnosed clinically and treated with antitoxin immediately. During follow-up, consecutive nerve stimulation was performed and significant incremental response was observed. There was an improvement in symptoms within 2 weeks, and in 5 or 6 weeks the symptoms had disappeared completely. Electrodiagnostic studies revealed that the findings turned to normal. The case showed that immediate antitoxin treatment is life-saving even the diagnosis of botulinum intoxication is based on clinical findings.

  3. Clinical efficacy of botulinum toxin type A injected by BellaVita in treatment of focal hyperhidrosis%A型肉毒毒素治疗局部多汗症的临床效果

    Institute of Scientific and Technical Information of China (English)

    陈凤超; 杨俊革; 夏秉成; 侯俊杰; 刘林嶓

    2016-01-01

    目的 探讨A型肉毒毒素治疗多汗症的临床效果.方法 观察28例多汗症患者132个部位治疗效果.用微碘-淀粉试验判定多汗症程度及范围.将A型肉毒毒素加入水光注射仪,单侧腋窝、手掌及脚掌注射剂量各50 U,额部30 U.注射后1周、2周和每月复查1次,随访8个月.每次随访根据微碘-淀粉试验的客观效果评价及主观效果评价得出分值,再进行综合效果评价.结果 注射前综合效果评价(1.34±3.94)分,A型肉毒毒素注射后1周综合效果评价(23.21±9.44)分,注射后1个月(92.41±11.95)分,2个月(98.21±5.60)分,3个月(95.98±5.94)分,4个月(86.61±10.17)分.与注射前比较,差异有统计学意义(P<0.05).之后效果慢慢减退,最长疗效维持可达8个月,综合效果评价(4.46±6.98)分,与注射前比较,差异无统计学意义(P>0.05).注射后6个月和注射后l周比较;差异无统计学意义(P>0.05).结论 A型肉毒毒素通过水光注射仪给药途径治疗局部多汗症见效快,疗效确切.%Objective To explore the new way of administration and clinical effect of botulinum toxin A in the treatment of focal hyperhidrosis.Methods The clinical efficacy was observed in 132 sites of 28 patients with focal hyperhidrosis,and the degree and range of focal hyperhidrosis were determined by the minor iodine-starch test.50 U of botulinum toxin A was injected in unilateral axillary,palms and soles with BellaVita instrument and 30 U for forehead.Each patient was followed-up in 1 week,2 weeks and every month after injection for 8 months.According to the results of the minor iodine-starch test the objective effect and evaluation score were obtained,and the comprehensive effect evaluation score was calculated with the objective effect evaluation score and the subjective effect evaluation score in each follow-up.Results The comprehensive effect evaluation score before injection of botulinum toxin A was 1.34±3.94,and that after injection was 23

  4. Novel Animal Defenses against Predation: A Snail Egg Neurotoxin Combining Lectin and Pore-Forming Chains That Resembles Plant Defense and Bacteria Attack Toxins

    Science.gov (United States)

    Ceolín, Marcelo; Ituarte, Santiago; Qiu, Jian-Wen; Sun, Jin; Fernández, Patricia E.; Heras, Horacio

    2013-01-01

    Although most eggs are intensely predated, the aerial egg clutches from the aquatic snail Pomacea canaliculata have only one reported predator due to unparalleled biochemical defenses. These include two storage-proteins: ovorubin that provides a conspicuous (presumably warning) coloration and has antinutritive and antidigestive properties, and PcPV2 a neurotoxin with lethal effect on rodents. We sequenced PcPV2 and studied whether it was able to withstand the gastrointestinal environment and reach circulation of a potential predator. Capacity to resist digestion was assayed using small-angle X-ray scattering (SAXS), fluorescence spectroscopy and simulated gastrointestinal proteolysis. PcPV2 oligomer is antinutritive, withstanding proteinase digestion and displaying structural stability between pH 4.0–10.0. cDNA sequencing and protein domain search showed that its two subunits share homology with membrane attack complex/perforin (MACPF)-like toxins and tachylectin-like lectins, a previously unknown structure that resembles plant Type-2 ribosome-inactivating proteins and bacterial botulinum toxins. The protomer has therefore a novel AB toxin combination of a MACPF-like chain linked by disulfide bonds to a lectin-like chain, indicating a delivery system for the former. This was further supported by observing PcPV2 binding to glycocalix of enterocytes in vivo and in culture, and by its hemaggutinating, but not hemolytic activity, which suggested an interaction with surface oligosaccharides. PcPV2 is able to get into predator’s body as evidenced in rats and mice by the presence of circulating antibodies in response to sublethal oral doses. To our knowledge, a lectin-pore-forming toxin has not been reported before, providing the first evidence of a neurotoxic lectin in animals, and a novel function for ancient and widely distributed proteins. The acquisition of this unique neurotoxic/antinutritive/storage protein may confer the eggs a survival advantage, opening new

  5. Quinolinic Acid: Neurotoxin or Oxidative Stress Modulator?

    Directory of Open Access Journals (Sweden)

    Lenka Kubicova

    2013-10-01

    Full Text Available Quinolinic acid (2,3-pyridinedicarboxylic acid, QUIN is a well-known neurotoxin. Consequently, QUIN could produce reactive oxygen species (ROS. ROS are generated in reactions catalyzed by transition metals, especially iron (Fe. QUIN can form coordination complexes with iron. A combination of differential pulse voltammetry, deoxyribose degradation and Fe(II autoxidation assays was used for explorating ROS formation in redox reactions that are catalyzed by iron in QUIN-Fe complexes. Differential pulse voltammetry showed an anodic shift of the iron redox potential if iron was liganded by QUIN. In the H2O2/FeCl3/ascorbic acid variant of the deoxyribose degradation assay, the dose-response curve was U-shaped. In the FeCl3/ascorbic acid variant, QUIN unambiguously showed antioxidant effects. In the Fe(II autoxidation assay, QUIN decreased the rate of ROS production caused by Fe(II oxidation. Our study confirms that QUIN toxicity may be caused by ROS generation via the Fenton reaction. This, however, applies only for unnaturally high concentrations that were used in attempts to provide support for the neurotoxic effect. In lower concentrations, we show that by liganding iron, QUIN affects the Fe(II/Fe(III ratios that are beneficial to homeostasis. Our results support the notion that redox chemistry can contribute to explaining the hormetic dose-response effects.

  6. Development of improved defined media for Clostridium botulinum serotypes A, B, and E.

    OpenAIRE

    Whitmer, M E; Johnson, E A

    1988-01-01

    The minimal nutritional growth requirements were determined for strains Okra B and Iwanai E, which are representatives of groups I and II, respectively, of Clostridium botulinum. These type B and E strains differed considerably in their nutrient requirements. The organic growth factors required in high concentrations by the Okra B strain (group I) were arginine and phenylalanine. Low concentrations (less than or equal to 0.1 g/liter) of eight amino acids (methionine, leucine, valine, isoleuci...

  7. Central antinociceptive activity of botulinum toxin A [Središnje antinociceptivno djelovanje botulinum toksina A

    OpenAIRE

    Matak, Ivica

    2015-01-01

    UVOD I CILJ ISTRAŽIVANJA: Botulinum toksin tipa A (BoNT/A), neurotoksin iz anaerobne bakterije Clostridium botulinum, je jedan od najpotentnijih bioloških toksina. Ulaskom u živčane terminale uzrokuje enzimsko cijepanje sinaptosomalnog proteina molekulske mase od 25 kDa (eng. synaptosomal-associated protein of 25 kDa; SNAP-25), što sprječava lučenje neurotransmitora. Intoksikacija organizma preko hrane ili infekcija sporama bakterija pri određenim uvjetima uzrokuje neuroparalitičku bolest bot...

  8. Clinical research of peripheral neurotomy and botulinum toxin type A in treatment of spastic cerebral palsy%周围神经缩窄术与A型肉毒毒素注射治疗痉挛型脑瘫的临床研究

    Institute of Scientific and Technical Information of China (English)

    钱洪玉; 王秀英; 李霞

    2013-01-01

    Objective To compare the effect and side-effect of peripheral neurotomy and botulinum toxin type A in the treatment of spastic cerebral palsy. Methods The 96 children with spastic cerebral palsy were randomly divided into three groups. They all received treatment of brain protein hydrolysate and normal function training. And the function training include 5 times every week for three months. On this condition, children of group A accepted peripheral neurotomy, and children of group B accepted botulinum toxin type A. Results After treatment, ADL and NIHSS of children in group A and B were improved than before. The effective rate of group A (93.75%) was obviously higher than group C(56.25%), and group B(90.63%) vice versa. The differences had statistics meaning(P0.05). Conclusion Peripheral neurotomies and botulinum toxin type A in treatment of spastic cerebral palsy are superior than conventional treatment. The two treatmens can enhance activities of daily living and improve levels of neurologic impairment.%目的:比较周围神经缩窄术与A型肉毒毒素注射治疗痉挛型脑瘫疗效及副作用,探讨痉挛性脑瘫的最适治疗方式及其效果。方法将118例患者随机分为A、B、C三组,3组患者均接受正规功能训练(每周5次,连续3个月以上), A组48例,给予周围神经缩窄术。B组28例,给予A型肉毒毒素注射治疗, C组42例,正规功能训练,(每周5次,连续3个月以上);观察对比3组患者的临床疗效及副作用。结果 A、B两组患者经治疗后日常生活能力、神经功能缺损程度评分明显改善;A组总有效率为93.75%, B组总有效率为92.86%,均高于C组(59.52%),差异有统计学意义(P0.05)。另外, A组有9例患者手术后肌力不同程度下降,4例患者肌肉痉挛不同程度复发,B组均出现暂时性不同程度局部疼痛。结论周围神经缩窄术与A型肉毒毒素注射治疗效果均优于常规治疗,可更好地改善患者神经缺损程度,提高

  9. Botulinum Toxin in Secondarily Nonresponsive Patients with Spasmodic Dysphonia.

    Science.gov (United States)

    Mor, Niv; Tang, Christopher; Blitzer, Andrew

    2016-09-01

    Chemodenervation with botulinum toxin (BoNT) has been effective and well tolerated for all types of dystonia for >30 years. We reviewed outcomes of our patients treated with BoNT serotype A (BoNT-A) for spasmodic dysphonia (SD) who became secondarily nonresponsive. We found that 8 of 1400 patients became nonresponsive to BoNT-A (0.57%), which is lower than the secondary nonresponse rate in other dystonias. After a cessation period, 4 of our patients resumed BoNT-A injections, and recurrence of immunoresistance was not seen in any of them. When compared with patients with other dystonias, patients with SD receive extremely low doses of BoNT. Small antigen challenge may explain the lower rate of immunoresistance and long-lasting efficacy after BoNT-A is restarted among secondary nonresponsive patients with SD. PMID:27143711

  10. Preparation and in Vitro Release of Self-Assembled Neurotoxin-Loaded Nanoparticles of Core-Shell Type%神经毒素自组装核壳型纳米粒的制备及其体外释药研究

    Institute of Scientific and Technical Information of China (English)

    柳琳; 徐陆忠; 何晓玮; 赵燕敏; 冯健; 李范珠

    2009-01-01

    目的 制备亲水性多肽类药物神经毒素的自组装核壳型纳米粒,并对其理化性质及体外释药特性进行考察.方法 以聚乙二醇一聚氰基丙烯酸乙酯嵌段共聚物(PEG-g-PECA)为载体,乳化聚合法制备神经毒素自组装核壳型纳米粒,采用正交实验优化制备工艺,制得的核壳型纳米粒通过透射电镜、Zeta电位/粒度分布仪考察理化性质,并用透析袋法分别研究其在pH 7.4和6.8的PBS缓冲液中的体外释药特性.结果 PEG-g-PECA能包埋亲水性多肽神经毒素,制备的神经毒素自组装核壳型纳米粒粒径为(89.6±8.9)mm,多分散系数为(0.110±0.003),包封率为(58.43±0.62)%,Zeta电位为(-38.81±0.47)mV;在pH 7.4和6.8的PBS缓冲液中的体外释药行为均符合Weibull方程,分别为lnln[1/(1-Q)]=0.474Int-1.612 1,r=0.994 6(pH 7.4)及lnln[1/(1-Q)]=0.351Int-0.827 1,r=0.970 8(pH 6.8).结论 以PEG-g-PECA为载体制备亲水性多肽类药物自组装核壳型纳米粒方法可行,所得纳米粒包封率较高,理化性质稳定,体外释药具有缓释制剂特征.%OBJECTIVE To prepare the self-assembled neurotoxin-loaded nanoparticles of core-shell type, and investigate its physicochemical properties and release behavior in vitro. METHODS The self-assembled neurotoxin-loaded nanoparticles of core-shell type were prepared with PEG-g-PECA by emulsion polymerization method. The orthogonal design was used to optimize the preparation technology. The transmission electron microscope(TEM) and Zeta sizer instrument were utilized to investigate the physicochemical properties of the nanoparticles, and the drug release behavior in PBS buffer at pH 7.4 and 6.8 in vitro were studied by dialysis method respectively. RESULTS The mean diameter of the nanoparticles were (89.6±8.9)nm with polydispersity index of (0.110±0.003). The entrapment efficiency was (58.43±0.62)%, and Zeta potential was (-38.81±0.47) inV. The in vitro release profiles of the nanoparticles in pH 7

  11. Botulinum Toxin in the Treatment of Facial Paralysis.

    Science.gov (United States)

    Mehdizadeh, Omid B; Diels, Jacqueline; White, William Matthew

    2016-02-01

    This article reviews the current literature supporting the use of botulinum toxin in producing symmetric facial features and reducing unwanted, involuntary movements. Methods, protocols, and adverse events are discussed. Additionally, the authors suggest that using botulinum toxin A therapy in postparalytic facial synkinesis can provide long-term results when used in conjunction with other treatment modalities.

  12. Effect of treatment with botulinum toxin on spasticity.

    OpenAIRE

    Das, T K; Park, D M

    1989-01-01

    Botulinum toxin, a product of Clostridium botulinum, produces presynaptic neuromuscular block by preventing release of acetylcholine from nerve endings. The toxin was injected directly into the skeletal muscles of six patients with severe spasticity due to stroke-related hemiplegia. It produced both subjective and objective improvement. The toxin injections were well tolerated and no significant side effect was reported.

  13. Practical guidance for CD management involving treatment of botulinum toxin: a consensus statement.

    Science.gov (United States)

    Albanese, Alberto; Abbruzzese, Giovanni; Dressler, Dirk; Duzynski, Wojciech; Khatkova, Svetlana; Marti, Maria Jose; Mir, Pablo; Montecucco, Cesare; Moro, Elena; Pinter, Michaela; Relja, Maja; Roze, Emmanuel; Skogseid, Inger Marie; Timerbaeva, Sofiya; Tzoulis, Charalampos

    2015-10-01

    Cervical dystonia is a neurological movement disorder causing abnormal posture of the head. It may be accompanied by involuntary movements which are sometimes tremulous. The condition has marked effects on patients' self-image, and adversely affects quality of life, social relationships and employment. Botulinum neurotoxin (BoNT) is the treatment of choice for CD and its efficacy and safety have been extensively studied in clinical trials. However, current guidelines do not provide enough practical information for physicians who wish to use this valuable treatment in a real-life setting. In addition, patients and physicians may have different perceptions of what successful treatment outcomes should be. Consequently, an international group of expert neurologists, experienced in BoNT treatment, met to review the literature and pool their extensive clinical experience to give practical guidance about treatment of CD with BoNT. Eight topic headings were considered: the place of BoNT within CD treatment options; patient perspectives and desires for treatment; assessment and goal setting; starting treatment with BoNT-A; follow-up sessions; management of side effects; management of non-response; switching between different BoNT products. One rapporteur took responsibility for summarising the current literature for each topic, while the consensus statements were developed by the entire expert group. These statements are presented here along with a discussion of the background information. PMID:25877834

  14. A novel neurotoxin from venom of the spider, Brachypelma albopilosum.

    Science.gov (United States)

    Zhong, Yunhua; Song, Bo; Mo, Guoxiang; Yuan, Mingwei; Li, Hongli; Wang, Ping; Yuan, Minglong; Lu, Qiumin

    2014-01-01

    Spiders have evolved highly selective toxins for insects. There are many insecticidal neurotoxins in spider venoms. Although a large amount of work has been done to focus on neurotoxicity of spider components, little information, which is related with effects of spider toxins on tumor cell proliferation and cytotoxicity, is available for Brachypelma albopilosum venom. In this work, a novel spider neurotoxin (brachyin) was identified and characterized from venoms of the spider, Brachypelma albopilosum. Brachyin is composed of 41 amino acid residues with the sequence of CLGENVPCDKDRPNCCSRYECLEPTGYGWWYASYYCYKKRS. There are six cysteines in this sequence, which form three disulfided bridges. The serine residue at the C-terminus is amidated. Brachyin showed strong lethal effects on American cockroaches (Periplaneta americana) and Tenebrio molitor (common mealbeetle). This neurotoxin also showed significant analgesic effects in mice models including abdominal writhing induced by acetic acid and formalin-induced paw licking tests. It was interesting that brachyin exerted marked inhibition on tumor cell proliferation.

  15. A novel neurotoxin from venom of the spider, Brachypelma albopilosum.

    Directory of Open Access Journals (Sweden)

    Yunhua Zhong

    Full Text Available Spiders have evolved highly selective toxins for insects. There are many insecticidal neurotoxins in spider venoms. Although a large amount of work has been done to focus on neurotoxicity of spider components, little information, which is related with effects of spider toxins on tumor cell proliferation and cytotoxicity, is available for Brachypelma albopilosum venom. In this work, a novel spider neurotoxin (brachyin was identified and characterized from venoms of the spider, Brachypelma albopilosum. Brachyin is composed of 41 amino acid residues with the sequence of CLGENVPCDKDRPNCCSRYECLEPTGYGWWYASYYCYKKRS. There are six cysteines in this sequence, which form three disulfided bridges. The serine residue at the C-terminus is amidated. Brachyin showed strong lethal effects on American cockroaches (Periplaneta americana and Tenebrio molitor (common mealbeetle. This neurotoxin also showed significant analgesic effects in mice models including abdominal writhing induced by acetic acid and formalin-induced paw licking tests. It was interesting that brachyin exerted marked inhibition on tumor cell proliferation.

  16. A novel neurotoxin from venom of the spider, Brachypelma albopilosum.

    Science.gov (United States)

    Zhong, Yunhua; Song, Bo; Mo, Guoxiang; Yuan, Mingwei; Li, Hongli; Wang, Ping; Yuan, Minglong; Lu, Qiumin

    2014-01-01

    Spiders have evolved highly selective toxins for insects. There are many insecticidal neurotoxins in spider venoms. Although a large amount of work has been done to focus on neurotoxicity of spider components, little information, which is related with effects of spider toxins on tumor cell proliferation and cytotoxicity, is available for Brachypelma albopilosum venom. In this work, a novel spider neurotoxin (brachyin) was identified and characterized from venoms of the spider, Brachypelma albopilosum. Brachyin is composed of 41 amino acid residues with the sequence of CLGENVPCDKDRPNCCSRYECLEPTGYGWWYASYYCYKKRS. There are six cysteines in this sequence, which form three disulfided bridges. The serine residue at the C-terminus is amidated. Brachyin showed strong lethal effects on American cockroaches (Periplaneta americana) and Tenebrio molitor (common mealbeetle). This neurotoxin also showed significant analgesic effects in mice models including abdominal writhing induced by acetic acid and formalin-induced paw licking tests. It was interesting that brachyin exerted marked inhibition on tumor cell proliferation. PMID:25329070

  17. Attempts to identify Clostridium botulinum toxin in milk from three experimentally intoxicated Holstein cows

    Science.gov (United States)

    Moeller, R.B.; Puschner, B.; Walker, R.L.; Rocke, T.E.; Smith, S.R.; Cullor, J.S.; Ardans, A.A.

    2009-01-01

    Three adult lactating Holstein cows were injected in the subcutaneous abdominal vein with 175 ng/kg of body weight of Clostridium botulinum type C toxin (451 cow median toxic doses) to determine if this botulinum toxin crosses the blood-milk barrier. Whole blood (in sodium heparin) and clotted blood serum samples were taken at 0 min, 10 min, and 3, 6, 9, and 12 h postinoculation. Milk samples were taken at 0 min and at 3, 6, 9 and 12 h postinoculation. All samples were tested for the presence of the toxin using the mouse bioassay and immunostick ELISA test. The immunostick ELISA identified the toxin in whole blood and the mouse bioassay identified the toxin in serum at all times examined in all 3 animals. Toxin was not identified by either detection method in milk samples collected from the 3 animals. From these results, it appears that Clostridium botulinum type C toxin does not cross from the blood to the milk in detectable concentrations. ?? American Dairy Science Association, 2009.

  18. Acute Radiation Disease : Cutaneous Syndrome and Toxic properties of Radiomimetics -Radiation Neurotoxins and Hematotoxins.

    Science.gov (United States)

    Popov, Dmitri; Maliev, Slava

    Cutaneous injury is an important complication of a general or local acute irradiation. A type of a skin and tissues lesions depends on a type, intensity, and period of irradiation. Also, the clinical picture, signs, and manifestations of the cutaneous syndrome depend on a type of the radiation toxins circulated in lymph and blood of irradiated mammals. Radiation Toxins were isolated from lymph of the mammals that were irradiated and developed different forms of the Acute Radiation Syndromes (ARS) -Cerebrovascular, Cardiovascular, Gastrointestinal, and Hematopoietic. Radiation Toxins can be divided into the two important types of toxins (Neu-rotoxins and Hematotoxins) or four groups. The effects of Radiation Neurotoxins include severe damages and cell death of brain, heart, gastrointestinal tissues and endothelial cells of blood and lymphatic vessels. The hematotoxicity of Hematotoxic Radiation Toxins includes lym-phopenia, leukopenia, thrombocytopenia, and anemia in the blood circulation and transitory lymphocytosis and leukocytosis in the Central Lymphatic System. In all cases, administration of the Radiomimetics (Radiation Toxins) intramuscularly or intravenously to healthy, radiation naive mammals had induced and developed the typical clinical manifestations of the ARS. In all cases, administration of Radiomimetics by subtoxic doses had demonstrated development of typical clinical signs of the cutaneous syndrome such as hair loss, erythema, swelling, desqua-mation, blistering and skin necrosis. In animal-toxic models, we have activated development of the local skin and tissue injury after injection of Radiation Toxins with cytoxic properties.

  19. 不同定位技术引导A型肉毒毒素注射治疗脑卒中后足下垂内翻的效果%Comparison of Location Guided by Palpation, Ultrasound and Electromyography in Botulinum Toxin Type A In-jection for Equinovarus in Stroke

    Institute of Scientific and Technical Information of China (English)

    欧海宁; 黄彬; 周凯欣; 黄卓群; 邓万溪; 卢伟焰

    2015-01-01

    Objective To compare the clinical effects of palpation, ultrasound and electromyography (EMG) guided injection of botuli-num toxin type A (BTX-A) on spastic equinovarus in stroke patients. Methods Stroke patients with equinovarus were randomly assigned to palpation-guided group (n=19), ultrasound-guided group (n=21) and EMG-guided group (n=19). All of the patients received injection of BTX-A 300 U in gastrocnemius, soleus and tibialis posterior, guided in their own ways. They were measured with modified Ashworth Scale (MAS), Physician Rating Scale (PRS), speed of gait, passive range of motion (PROM) of ankle dorsiflexion and eversion before and 2 weeks, 4 weeks, 8 weeks, and 12 weeks after injection. Results Compared with the baseline, all the indexes significantly improved after in-jection in all the groups (P0.05).结论 三种定位技术引导下注射BTX-A均能改善患者痉挛和步行能力,超声引导比肌电图引导在改善足内翻方面稍有优势,均优于徒手定位.

  20. Botulinum toxin in the treatment of sialorrhea

    Directory of Open Access Journals (Sweden)

    Svetel Marina

    2009-01-01

    Full Text Available Background/Aim. Botulinum toxin-A (BTX-A is known to block the release of acetylcholine from motor and autonomic nerve terminals and may significantly decrease saliva production when injected intraglandulary. The aim of this study was to evaluate effects of BTX-A injections in the treatment of disabling sialorrhea in various neurological disorders. Methods. This study included 19 consecutive patients with significant sialorrhea associated with various neurological disorders. Out of them 13 patients were with Parkinson's disease, two with pantothenate kinase-associated neurodegeneration, two with multiple system atrophy, one with Wilson's disease, and one patient with postoperative sialorrhea. Botulinum toxin-A (Dysport®, Ipsen Pharma was injected into the parotid glands with (n = 7 patients or without (n = 12 patients ultrasound guidance. All the patients were scored before and after the treatment and in weekly intervals thereafter using the salivation item of the part II (Activities of Daily Living of the Unified Parkinson's Disease Rating Scale (UPDRS. Results. Thirteen patients (68% reported beneficial effect of BTX-A injection, while 6 of them (32% had no response at all. The sialorrhea scores before and after the treatment were 3.1 ± 0.1 (range 2-4 and 1.8 ± 0.1 (range 0- 3, respectively (t = 5.636; p < 0.001. There was no difference in the magnitude of response between the groups with (t = 4.500; p = 0.004 and without (t = 3.674; p = 0.005 ultrasound control of injection sites. Adverse effects were registered in 5 patients (26%. Conclusions. Botulinum toxin-A injections to easily accessible parotid glands, without necessity for ultrasound guidance, are safe and efficaceous treatment for sialorrhea in different neurological disorders.

  1. Ultrasound-guided botulinum toxin injections

    Directory of Open Access Journals (Sweden)

    S. E. Khatkova

    2016-01-01

    Full Text Available One of the key conditions for achieving the desirable result during botulinum toxin therapy for muscular dystonia, spasticity, and other diseases accompanied by spasm, pain, and autonomic dysfunction (dystonias, spasticity, etc. is the proper administration of the agent into the muscles directly involved in the pathological process. The exact entry of botulinum toxin into the target muscles is essential for successful and safe treatment because its injection into a normal muscle may cause side effects. The most common errors are the incorrect depth and incorrect direction of a needle on insertion. Therefore, the exact injection of the agent particularly into the shallow and deep muscles is a difficult task even for an experienced specialist and requires the use of controlling methods.The European Consensus on Botulinum Toxin Therapy points out that various injection techniques are needed for the better identification of necessary muscles. However, there are currently no reports on the clear advantage of any technique. In our country, injections using palpation and anatomical landmarks have been widely used in routine practice so far; electromyographic monitoring and electrostimulation have been less frequently applied. In recent years, the new method ultrasound-guided injection has continued to grow more popular. This effective, accessible, and easy-to-use method makes it possible to manage a real-time injection process and to ensure the exact entry of the agent into the muscle. This paper is dedicated to a comparative analysis of different injection methods and to a description of the ultrasound-guided technique and its advantages over others. 

  2. Botulinum Toxin Physiology in Focal Hand and Cranial Dystonia

    Directory of Open Access Journals (Sweden)

    Barbara Illowsky Karp

    2012-11-01

    Full Text Available The safety and efficacy of botulinum toxin for the treatment of focal hand and cranial dystonias are well-established. Studies of these adult-onset focal dystonias reveal both shared features, such as the dystonic phenotype of muscle hyperactivity and overflow muscle contraction and divergent features, such as task specificity in focal hand dystonia which is not a common feature of cranial dystonia. The physiologic effects of botulinum toxin in these 2 disorders also show both similarities and differences. This paper compares and contrasts the physiology of focal hand and cranial dystonias and of botulinum toxin in the management of these disorders.

  3. Cosmetic Effect of Botulinum Toxin In Focal Hyperhydrosis

    Directory of Open Access Journals (Sweden)

    Jain S

    2005-01-01

    Full Text Available Hyperhydrosis of axillae, palm and sole is not a very uncommon problem. It leads to great embarrassment and considerable emotional stress to the individuals. Botulinum toxins prevent the release of acetylcholine at nerve terminals, therefore, reduces sweat secretion. Six patients of axillary and 4 patients of palmer and planter hyperhydrosis were treated with botulinum toxin. All patients experienced relatively satisfactory reduction of hyperhydrosis for period ranging between 4-7 months. No adverse effects were observed. Botulinum toxin therefore can be considered as an effective treatment in focal hyperhydrosis.

  4. Botulinum toxin for treatment of glandular hypersecretory disorders.

    LENUS (Irish Health Repository)

    Laing, T A

    2012-02-03

    SUMMARY: The use of botulinum toxin to treat disorders of the salivary glands is increasing in popularity in recent years. Recent reports of the use of botulinum toxin in glandular hypersecretion suggest overall favourable results with minimal side-effects. However, few randomised clinical trials means that data are limited with respect to candidate suitability, treatment dosages, frequency and duration of treatment. We report a selection of such cases from our own department managed with botulinum toxin and review the current data on use of the toxin to treat salivary gland disorders such as Frey\\'s syndrome, excessive salivation (sialorrhoea), focal and general hyperhidrosis, excessive lacrimation and chronic rhinitis.

  5. Botulinum Toxin Injection for Spastic Scapular Dyskinesia After Stroke

    Science.gov (United States)

    Hou, Saiyun; Ivanhoe, Cindy; Li, Sheng

    2015-01-01

    Abstract Spastic scapular dyskinesia after stroke is rare, which causes impaired shoulder active range of motion (ROM). To date, there has been no report about botulinum toxin injection to spastic periscapular muscles. This study presents botulinum toxin A injection for management of spastic periscapular muscles after stroke in 2 cases. This is a retrospective study of 2 cases of spastic scapular dyskinesia after stroke. Spasticity of periscapular muscles including rhomboid and lower trapezius was diagnosed by physical examination and needle electromyographic study. Botulinum toxin was injected into the spastic periscapular muscles under ultrasound imaging guidance. During the 3-week follow-up visit after injection, both patients showed increased shoulder active ROM, without any sign of scapular destabilization. The results suggest that botulinum toxin injection to spastic periscapular muscles can increase shoulder active ROM without causing scapular destabilization in patients with poststroke spastic scapular dyskinesia. PMID:26266368

  6. Botulinum Toxin Physiology in Focal Hand and Cranial Dystonia

    OpenAIRE

    Barbara Illowsky Karp

    2012-01-01

    The safety and efficacy of botulinum toxin for the treatment of focal hand and cranial dystonias are well-established. Studies of these adult-onset focal dystonias reveal both shared features, such as the dystonic phenotype of muscle hyperactivity and overflow muscle contraction and divergent features, such as task specificity in focal hand dystonia which is not a common feature of cranial dystonia. The physiologic effects of botulinum toxin in these 2 disorders also show both similarities an...

  7. Botulinum Toxin Injection for Spastic Scapular Dyskinesia After Stroke

    OpenAIRE

    Hou, Saiyun; Ivanhoe, Cindy; Li, Sheng

    2015-01-01

    Abstract Spastic scapular dyskinesia after stroke is rare, which causes impaired shoulder active range of motion (ROM). To date, there has been no report about botulinum toxin injection to spastic periscapular muscles. This study presents botulinum toxin A injection for management of spastic periscapular muscles after stroke in 2 cases. This is a retrospective study of 2 cases of spastic scapular dyskinesia after stroke. Spasticity of periscapular muscles including rhomboid and lower trapeziu...

  8. Antagonism of the paralysis produced by botulinum toxin in the rat. The effects of tetraethylammonium, guanidine and 4-aminopyridine.

    Science.gov (United States)

    Lundh, H; Leander, S; Thesleff, S

    1977-05-01

    The injection of botulinum toxin type A into the hind-leg of adult rats causes complete paralysis of the leg lasting for several weeks. In the extensor digitorum longus (EDL) muscle transmitter release is reduced to a level of less than 1% of normal. Tetraethylammonium (TEA) and guanidine in concentrations of about 3 mM restore, in EDL muslces in vitro, neuromuscular transmission to about the normal level, provided that the external calcium concentration is 4 mM or higher. 4-Aminopyridine (4-AP) has similar restorative effect but is about 20-30 times more potent. Unlike TEA and guanidine, 4-AP is effective when the ambient calcium concentration is 2 mM; this drug is therefore also active in vivo. The intravenous injection of 4-AP (5 mg/kg body weight) restores neuromuscular transmission from complete paralysis by botulinum toxin to a normal level as shown by the recording of almost normal twitch and tetanic tensions in the EDL muscle. In rats paralysed by a lethal dose of botulinum toxin, the intraperitoneal administration of 4-AP restores general motor activity, the effect lasting 1-2 hours. A study of the effects of these drugs on spontaneous and evoked transmitter release suggests that all three compounds increase the level of free calcium inside the nerve terminals. In botulinum poisoning the transmitter release mechanism appears to be intact, but a reduced sensitivity to calcium has been shown (Cull-Candy et al. 1976), and this could explain why the drugs restore evoked transmitter release in botulinum poisoning. PMID:194021

  9. Prioritizing drug targets in Clostridium botulinum with a computational systems biology approach.

    Science.gov (United States)

    Muhammad, Syed Aun; Ahmed, Safia; Ali, Amjad; Huang, Hui; Wu, Xiaogang; Yang, X Frank; Naz, Anam; Chen, Jake

    2014-07-01

    A computational and in silico system level framework was developed to identify and prioritize the antibacterial drug targets in Clostridium botulinum (Clb), the causative agent of flaccid paralysis in humans that can be fatal in 5 to 10% of cases. This disease is difficult to control due to the emergence of drug-resistant pathogenic strains and the only available treatment antitoxin which can target the neurotoxin at the extracellular level and cannot reverse the paralysis. This study framework is based on comprehensive systems-scale analysis of genomic sequence homology and phylogenetic relationships among Clostridium, other infectious bacteria, host and human gut flora. First, the entire 2628-annotated genes of this bacterial genome were categorized into essential, non-essential and virulence genes. The results obtained showed that 39% of essential proteins that functionally interact with virulence proteins were identified, which could be a key to new interventions that may kill the bacteria and minimize the host damage caused by the virulence factors. Second, a comprehensive comparative COGs and blast sequence analysis of these proteins and host proteins to minimize the risks of side effects was carried out. This revealed that 47% of a set of C. botulinum proteins were evolutionary related with Homo sapiens proteins to sort out the non-human homologs. Third, orthology analysis with other infectious bacteria to assess broad-spectrum effects was executed and COGs were mostly found in Clostridia, Bacilli (Firmicutes), and in alpha and beta Proteobacteria. Fourth, a comparative phylogenetic analysis was performed with human microbiota to filter out drug targets that may also affect human gut flora. This reduced the list of candidate proteins down to 131. Finally, the role of these putative drug targets in clostridial biological pathways was studied while subcellular localization of these candidate proteins in bacterial cellular system exhibited that 68% of the

  10. Neutralizing Antibody and Botulinum Toxin Therapy: A Systematic Review and Meta-analysis.

    Science.gov (United States)

    Fabbri, Margherita; Leodori, Giorgio; Fernandes, Ricardo M; Bhidayasiri, Roongroj; Marti, Maria Jose; Colosimo, Carlo; Ferreira, Joaquim J

    2016-01-01

    The formation of neutralizing antibodies (NAbs) directed specifically against the active neurotoxin part of the botulinum neurotoxin (BoNT) complex is often cited as a major cause of secondary non-responsiveness (SnR) to treatment. This systematic and meta-analytic review evaluates the frequency of NAbs among patients treated with BoNT therapy for any clinical indication. A comprehensive database search strategy was designed to retrieve relevant clinical data from the published literature up to April 2013. All English-language publications that analyzed NAbs prevalence in more than ten patients were included, regardless of BoNT formulation, assay method, and study design. For the meta-analysis, patients were divided into three categories: secondary nonresponse (SnR) patients, clinically responding patients and all patients, independently of BoNT responsiveness. The meta-analysis included 61 studies reporting data for 8525 patients; 4972 dystonic patients, 1170 patients with spasticity, 294 patients with urologic indications, 396 patient with hyperhidrosis, 1659 patients with glabellar line, and 34 patients with hypersalivation. Among the ‘‘all patients’’ group NAbs frequency was 20%for dystonia, 5.9%for spasticity, and 2.7% for urologic patients and 1.1% for other conditions. The prevalence of NAbs was lower (3.5%) among clinically responding patients and higher in 53.5%SnR patients. About a half of patients with SnR do not have NAbs. NAbs was high among patients treated with RIMA but it was not associated with clinical non-responsiveness. Meta-analysis of the frequency of NAbs and SnR are limited by the heterogeneity of study design and reported outcomes. Indeed the analysis of several factors that can influence the development of NAbs, i.e.,MHCof patients, frequency and site of injection, injection technique, cumulative dose, and toxin denaturation, was not specifically evaluated due to the paucity and heterogeneity of data. The identification of all

  11. A型肉毒毒素对P物质所致大鼠胃体、胃底离体平滑肌收缩的抑制作用%Inhibitory effect of botulinum toxin type A on SP-induced rat smooth muscle contractility of gastric body and gastric fundus in vitro

    Institute of Scientific and Technical Information of China (English)

    周媛媛; 李超彦; 侯一平

    2012-01-01

    目的 观察A型肉毒毒素(botulinum toxin type A,BTX-A)对P物质(substance P,SP)所致肌条收缩的影响,探讨BTX-A在SP与NK1受体结合过程中可能存在的作用机制.方法 取大鼠胃体、胃底平滑肌制备肌条并随机分为对照组、SP组、SP+ APTL-SP(NK1受体拮抗剂)组、BTX-A组、BTX-A+ SP组、SP+ BTX-A组,采用Biolap420E生物机能实验系统记录肌条收缩数据.结果 SP增加胃体平滑肌自发性收缩张力及振幅、胃底平滑肌自发性收缩张力(P均<0.01);APTL-SP降低SP引发的胃体、胃底平滑肌收缩张力(P<0.01); BTX-A作用后的胃体、胃底平滑肌条振幅降低(P均<0.01).BTX-A降低SP引发的胃体(P<0.05,P<0.01)、胃底(P均<0.01)平滑肌自发性收缩张力及振幅;SP对BTX-A作用后的胃体、胃底平滑肌收缩能力未产生增强作用.结论 SP可增强胃体、胃底平滑肌收缩能力,而BTX-A可抑制SP对胃体、胃底平滑肌的收缩作用.%Objective To observe the effect of botulinum toxin type A (BTX-A) on the SP-induced smooth muscle contractility of gastric body and gastric fundus, so as to investigate the role of BTX-A in the binding between SP and NKi receptor. Methods Muscle strips were prepared from gastric body and gastric fundus and were randomly divided into control group, SP group, SP+APTL-SP (NK, receptor antagonist) group, BTX-A group, BTX-A+SP group, and SP+BTX-A group. The contractility data were recorded by physiological experimental system of Biolap420E. Results SP significantly enhanced the tension and amplitude of gastric body contractility and the tension of gastric fundus contractility (P<0. 01). APTL-SP signficantly inhibited SP-induced smooth muscle contractility tension in gastric body and gastric fundus (P<0. 01). BTX-A significantly inhibited the smooth muscle contractility amplitude in gastric body and gastric fundus (P<0. 01). BTX-A significantly inhibited SP-induced smooth muscle contractility, including the

  12. Acute angle-closure glaucoma following botulinum toxin injection for blepharospasm.

    OpenAIRE

    Corridan, P.; Nightingale, S; Mashoudi, N.; Williams, A C

    1990-01-01

    Botulinum toxin inhibits acetylcholine release and therefore could cause mydriasis. We report a case of acute angle-closure glaucoma which occurred shortly after a series of injections of botulinum toxin round the eyelids for blepharospasm.

  13. FDA Approves First Botulism Antitoxin for Use in Neutralizing All Seven Known Botulinum Nerve Toxin Serotypes

    Science.gov (United States)

    ... Antitoxin for use in neutralizing all seven known botulinum nerve toxin serotypes Product to be stored in Strategic National ... antibody fragments that neutralize all of the seven botulinum nerve toxin serotypes known to cause botulism. Botulism is a ...

  14. Botulinum Toxin to Treat Neurogenic Bladder.

    Science.gov (United States)

    Smith, Christopher P; Chancellor, Michael B

    2016-02-01

    Alteration in neural control from suprapontine areas to the nerves innervating the bladder can lead to bladder dysfunction and the development of a neurogenic bladder (NGB). Patients with NGB often suffer from urinary incontinence, which can lead to adverse events such as urinary tract infections and decubiti, in addition to creating a large care burden for family members or healthcare providers and significantly impairing patient quality of life. The common failure of anticholinergic medications has spurned the development of second-line treatments, including the use of botulinum toxin. OnabotulinumtoxinA (onaBoNT-A; BOTOX, Allergan, Inc.) was approved by the U.S. Food and Drug Administration (FDA) in 2011 to treat neurogenic detrusor overactivity in patients with urinary incontinence resulting from a NGB. In this review the authors summarize pertinent results from key trials leading to FDA approval of onaBoNT-A as well as more recent long-term data.

  15. What is the risk of aluminium as a neurotoxin?

    Science.gov (United States)

    Exley, Christopher

    2014-06-01

    Aluminium is neurotoxic. Its free ion, Al(3+) (aq), is highly biologically reactive and uniquely equipped to do damage to essential cellular (neuronal) biochemistry. This unequivocal fact must be the starting point in examining the risk posed by aluminium as a neurotoxin in humans. Aluminium is present in the human brain and it accumulates with age. The most recent research demonstrates that a significant proportion of individuals older than 70 years of age have a potentially pathological accumulation of aluminium somewhere in their brain. What are the symptoms of chronic aluminium intoxication in humans? What if neurodegenerative diseases such as Alzheimer's disease are the manifestation of the risk of aluminium as a neurotoxin? How might such an (outrageous) hypothesis be tested?

  16. Cerebrovascular Acute Radiation Syndrome : Radiation Neurotoxins, Mechanisms of Toxicity, Neuroimmune Interactions.

    Science.gov (United States)

    Popov, Dmitri; Maliev, Slava

    Introduction: Cerebrovascular Acute Radiation Syndrome (CvARS) is an extremely severe in-jury of Central Nervous System (CNS) and Peripheral Nervous System (PNS). CvARS can be induced by the high doses of neutron, heavy ions, or gamma radiation. The Syndrome clinical picture depends on a type, timing, and the doses of radiation. Four grades of the CvARS were defined: mild, moderate, severe, and extremely severe. Also, four stages of CvARS were developed: prodromal, latent, manifest, outcome -death. Duration of stages depends on the types, doses, and time of radiation. The CvARS clinical symptoms are: respiratory distress, hypotension, cerebral edema, severe disorder of cerebral blood microcirculation, and acute motor weakness. The radiation toxins, Cerebro-Vascular Radiation Neurotoxins (SvARSn), determine development of the acute radiation syndrome. Mechanism of action of the toxins: Though pathogenesis of radiation injury of CNS remains unknown, our concept describes the Cv ARS as a result of Neurotoxicity and Excitotoxicity, cell death through apoptotic necrosis. Neurotoxicity occurs after the high doses radiation exposure, formation of radiation neuro-toxins, possible bioradicals, or group of specific enzymes. Intracerebral hemorrhage can be a consequence of the damage of endothelial cells caused by radiation and the radiation tox-ins. Disruption of blood-brain barrier (BBB)and blood-cerebrospinal fluid barrier (BCFB)is possibly the most significant effect of microcirculation disorder and metabolic insufficiency. NMDA-receptors excitotoxic injury mediated by cerebral ischemia and cerebral hypoxia. Dam-age of the pyramidal cells in layers 3 and 5 and Purkinje cell layer the cerebral cortex , damage of pyramidal cells in the hippocampus occur as a result of cerebral ischemia and intracerebral bleeding. Methods: Radiation Toxins of CV ARS are defined as glycoproteins with the molec-ular weight of RT toxins ranges from 200-250 kDa and with high enzymatic activity

  17. Adaptive Simulated Annealing Based Protein Loop Modeling of Neurotoxins

    Institute of Scientific and Technical Information of China (English)

    陈杰; 黄丽娜; 彭志红

    2003-01-01

    A loop modeling method, adaptive simulated annealing, for ab initio prediction of protein loop structures, as an optimization problem of searching the global minimum of a given energy function, is proposed. An interface-friendly toolbox-LoopModeller in Windows and Linux systems, VC++ and OpenGL environments is developed for analysis and visualization. Simulation results of three short-chain neurotoxins modeled by LoopModeller show that the method proposed is fast and efficient.

  18. Coupling the Torpedo Microplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

    OpenAIRE

    Aráoz, Rómulo; Ramos, Suzanne; Pelissier, Franck; Guérineau, Vincent; Benoit, Evelyne; Vilariño, Natalia; Botana, Luis M.; Zakarian, Armen; Molgó, Jordi

    2012-01-01

    Cyclic imine neurotoxins constitute an emergent family of neurotoxins of dinoflagellate origin that are potent antagonists of nicotinic acetylcholine receptors. We developed a target-directed functional method based on the mechanism of action of competitive agonists/antagonists of nicotinic acetylcholine receptors for the detection of marine cyclic imine neurotoxins. The key step for method development was the immobilization of Torpedo electrocyte membranes rich in nicotinic acetylcholine rec...

  19. Comparison between the effects of location by manual palpation and ultrasound guidance in botulinum toxin type A injection techniques for stroke patients' spastic equinovarus%超声引导和徒手肌肉定位法用于A型肉毒毒素治疗脑卒中患者痉挛性足下垂内翻的临床效果

    Institute of Scientific and Technical Information of China (English)

    欧海宁; 沈建虹; 陈红霞; 詹乐昌; 李梅

    2011-01-01

    Objective: To compare the clinical outcome of two different location techniques: manual palpation guidance and ultrasound guidance, for botulinum toxin type A injection into calf muscles in the treatment of spastic equinovarus in stroke patients.Method: Forty stroke patients with equinovarus were randomly assigned into two groups. All patients of both groups received injection of 300U botulinum toxin A in gastrocnemius, soleus and tibialis posterior muscles. The group A(n=18) was applied manual palpation-guided technique for location of injection. The group B(n=20) was applied ultrasound-guided technique for location of injection. Modified Ashworth scale(MAS), physician rating scale (PRS), speed of gait, passive range of motion(PROM) of ankle dorsiflexion and eversion were measured at baseline, and the 2nd, 4th, 8th, 12th week after treatment.Result: In each group, after treatment, the measurement indexes of MAS, gait pattern scale, speed of gait, PROM of ankle dorsiflexion and eversion all improved significantly(P<0.05). Group B improved significantly in MAS,PRS,speed of gait, PROM of ankle dorsiflexion and eversion at the 4th week(P<0.05), in MAS and PROM of ankle dorsiflexion at the 8th week(P<0.05), in MAS, speed of gait and PROM of ankle eversion at the 12th week comparing with group A(P<0.05). Comparing the trend of variations of measurement indexes during follow-up periods in two groups there was no statistical difference.Conclusion: The correction of equinovarus in stroke patients could be obtained by injecting botulinum toxin type A. Ultrasound-guided technique was considered to be a valid alternative technique for location of injection, which got superior clinical results than manual palpation-guided technique in some respects.%目的:比较超声引导下注射和徒手肌肉定位法在肉毒毒素注射治疗脑卒中患者足下垂内翻的临床效果.方法:46例符合纳人标准的脑卒中后足下垂内翻

  20. Feasibility of a Day-Camp Model of Modified Constraint-Induced Movement Therapy with and without Botulinum Toxin A Injection for Children with Hemiplegia

    Science.gov (United States)

    Eliasson, Ann-Christin; Shaw, Karin; Ponten, Eva; Boyd, Roslyn; Krumlinde-Sundholm, Lena

    2009-01-01

    The objective of the study was to investigate the feasibility of modified constraint-induced (CI) therapy provided in a 2-week day-camp model with and without intramuscular botulinum toxin type A (BoNT-A) injections for children with congenital cerebral palsy. Sixteen children with congenital hemiplegia, Manual Ability Classification System (MACS)…

  1. The effect of neurotoxin on rabies virus binding to mouse neuroblastoma cells.

    Science.gov (United States)

    Briggs, D J; Phillips, R M

    1991-08-01

    Mouse neuroblastoma cells were exposed to alpha bungarotoxin, a neurotoxin known to inhibit rabies virus binding to the nicotinic acetylcholine receptor located at the neuromuscular junction in muscle tissue. The total amount of 3H-CVS virus that bound to neurotoxin treated cells was separated into specific and non-specific binding using a cold competition assay. Comparison of untreated and neurotoxin treated cells demonstrated that the majority of cell-associated virus in untreated cells was of a specific nature whereas the majority of the cell-associated virus in neurotoxin treated cells was due to non-specific binding.

  2. Rating scales for cervical dystonia: a critical evaluation of tools for outcome assessment of botulinum toxin therapy.

    Science.gov (United States)

    Jost, Wolfgang H; Hefter, Harald; Stenner, Andrea; Reichel, Gerhard

    2013-03-01

    Botulinum neurotoxin is the therapy of choice for all forms of cervical dystonia (CD), but treatment regimens still vary considerably. The interpretation of treatment outcome is mainly based on the clinical experience and on the scientific value of the rating scales applied. The aim of this review is to describe the historical development of rating scales for the assessment of CD and to provide an appraisal of their advantages and drawbacks. The Tsui score and the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) have been widely employed in numerous clinical studies as specific instruments for CD. The obvious advantage of the Tsui score is its simplicity so that it can be easily implemented in clinical routine. The TWSTRS allows a more sophisticated assessment of functional features of CD, but only the Tsui score includes a rating for tremor. Other benefits of the TWSTRS are the disability and pain subscales, but despite its value in clinical trials, it might be too complex for routine clinical practice. None of the rating scales used at present has been rigorously tested for responsiveness to detect significant changes in clinical status after therapeutic interventions. Moreover, clinical data support a new classification of CD leading to a differentiation between head and neck subtypes. As the current rating scales are not able to cover all these aspects of the disorder, further research is needed to develop a valid and reliable instrument which considers the most current classification of CD.

  3. Bupivacaine and botulinum toxin to treat comitant strabismus

    Directory of Open Access Journals (Sweden)

    Luisa Moreira Hopker

    2012-04-01

    Full Text Available PURPOSE: To evaluate the change in ocular motility and muscle thickness measured with ultrasonography after intramuscular injection of bupivacaine and botulinum toxin A. METHODS: Eight patients (five female were enrolled to measure ocular motility prior and 1, 7, 30 and 180 days after one injection of 2 ml of 1.5% bupivacaine and 2.5 U of botulinum toxin A in agonist and antagonist muscles, respectively, of eight amblyopic eyes. Muscle thickness was measured prior and on days 1, 7 and 30 after injection using 10-MHz ultrasonography (eyelid technique. RESULTS: Mean change in alignment was 10 prism diopters after 180 days (n=6. An average increase of 1.01 mm in muscle thickness was observed after 30 days of bupivacaine injection and 0.28 mm increase was observed after botulinum toxin A injection, as measured by ultrasonography. Lateral rectus muscles injected with bupivacaine had a mean increase of 1.5 mm in muscle thickness. CONCLUSION: In this study, a change in ocular motility was observed after 180 days of intramuscular injection of bupivacaine and botulinum toxin in horizontal extraocular muscles. Overall, there was an increase of muscle thickness in both botulinum toxinum A and bupivacaine injected muscles after 30 days of injection when measured by ultrasonography. This change was more pronounced on lateral rectus muscles after bupivacaine injection.

  4. Treatment of displaced mandibular condylar fracture with botulinum toxin A.

    Science.gov (United States)

    Akbay, Ercan; Cevik, Cengiz; Damlar, Ibrahim; Altan, Ahmet

    2014-04-01

    The aim of this case report is to discuss the effect on condylar reduction of botulinum toxin A treatment used in a child with displaced fracture at condylar neck of mandible. A 3-years old boy was admitted to our clinic for incomplete fracture of mandibular symphysis and displaced condylar fracture at the left side. An asymmetrical occlusal splint with intermaxillary fixation was used instead of open reduction and internal fixation because of incomplete fracture of symphysis and possible complications of condyle surgery. However, it was observed that condylar angulation persisted despite this procedure. Thus, botulinum toxin A was administered to masseter, temporalis and pterygoideus medialis muscles. At the end of first month, it was seen that mandibular condyle was almost completely recovered and that fusion was achieved. In conclusion, Botulinum A toxin injection aiming the suppression of masticatory muscle strength facilitates the reduction in the conservative management of displaced condyle in pediatric patients. PMID:24156980

  5. [Treatment of blepharospasm with botulinum toxin].

    Science.gov (United States)

    Pikielny, R T; Micheli, F E; Fernández Pardal, M M; Casas Parera, I; Giannaula, R J; Gatto, M

    1990-01-01

    Blepharospasm is a relatively frequent cranial dystonia which may be seen either alone or related to orofacial-mandibular dystonia (Meige's syndrome). In its maximum degree it can cause functional blindness.Twelve patients with blepharospasm (4 essential and 8 Meige's syndrome) who had been previously treated unsuccessfully with drugs (trihexyphenidyl, biperiden, carbamazepine, lithium, baclofen, lisuride, imipramine, clonazepam and butyrophenones) were treated for 12 months with periocular injections of botulinum toxin (BOTOX). A "low" dose of 12,5 U per eye was employed. With this dose, eleven out of twelve patients experienced significant improvement which lasted from five to fifteen weeks. The only nonresponder obtained complete relief upon duplicating the dose. The only side effect was uni or bilateral ptosis in six patients which improved completely in seven to twenty one days. One patient developed a peripheral facial palsy with complete remission in nineteen days. No systemic side effects were noted. There was only one desertion from this study due to depression enhanced by prolonged (21 days) ptosis. All patients (including the deserter) agreed that treatment with BOTOX provided more relief than any other previous therapeutic method. Our results confirm those obtained by others but a more prolonged study is needed to better evaluate long term effects. PMID:2101846

  6. Clinical Efficacy of Ultrasound-guided Botulinum Toxin Type A Injection and Muscle Electricity Biofeedback Treatment for Patients with Spasticity in Lower Limb Muscles%彩超引导下肉毒毒素注射和肌电生物反馈治疗下肢肌肉痉挛的疗效观察

    Institute of Scientific and Technical Information of China (English)

    陈华先; 丁旭东; 张贵斌; 黄瓅; 宋金辉; 洪艳; 王汉

    2012-01-01

    Objective: To evaluate the clinical effect of ultrasound-guided botulinum toxin type A (BTX-A) injection and muscle electricity biofeedback treatment for patients with spastici-ty in the lower limb muscles after cerebral infarction. Methods: Thirty-six patients with lower limb muscle Spasms after cerebral infarction were randomly divided into control group and the treatment group with 18 cases in the each group. The ultrasound-guided BTX-A injection was applied to the both groups whereas muscle electricity biofeedback treatment was performed additional to those in the treatment group after 24 hours. The outcome was assessed by modified Ash-worth scale (MAS). Results: The recovery of Muscle spasms in the electricity biofeedback treatment group was better than that in the control group. Conclusion: The ultrasound guided BTX-A injection and muscle electricity biofeedback treatment are effective for treatingt patients with lower limb muscle spasticity after cerebral infarction.%目的:观察采用彩超引导下注射A型肉毒毒素(BTX-A)和肌电生物反馈治疗脑梗死后下肢肌肉痉挛的临床疗效.方法:选择36例脑梗死偏瘫后下肢肌肉痉挛患者,随机分成治疗组及对照组各18例.2组均对下肢痉挛肌群进行分点注射BTX-A,治疗组24 h后给予肌电生物反馈治疗6周,采用改良Ashworth评分(MAS)、步长、步速评估2组疗效.结果:治疗组治疗6周后MAS评分、步长、步速优于对照组(P<0.05).结论:采用彩超引导下多位点注射BTX-A结合肌电生物反馈治疗脑梗死后下肢肌肉痉挛有明显疗效.

  7. A型肉毒毒素降低偏头痛大鼠颈静脉血、脑干和三叉神经节的P物质含量%Botulinum toxin type A reducing SP immunoreactivity in jugular plasma,brain stem and ganglion nervi trigemini in migraine model rats triggered by nitroglycerin

    Institute of Scientific and Technical Information of China (English)

    张晓英; 侯一平; 宋焱峰; 王荫椿

    2006-01-01

    目的:通过观察A型肉毒毒素(botulinum toxin type A,BTX-A)对硝酸甘油引发的偏头痛大鼠颈静脉血浆、脑干和三叉神经节中P物质(substance P,SP)含量变化,探讨BTX-A止痛机理.方法:将大鼠分为对照组(n=5)和偏头痛模型组(n=15).偏头痛模型组造模后分别皮下注 0.9% NaCl(n=5)、BTX-A 5 U·kg-1 (n=5)、BTX-A 10 U·kg-1 (n=5) 6 d 后,应用放射免疫检测法测定颈静脉血浆、脑干和三叉神经节中P物质的含量.结果:动物皮下注射硝酸甘油后出现偏头痛症状,例如双耳发红、前肢频繁搔头、爬笼次数增多,其评分与对照组相比有明显差异(P<0.01).生理盐水组颈静脉血浆、脑干和三叉神经节中SP含量与对照组比较显著升高(P<0.01);BTX-A 5 U·kg-1组、BTX-A 10 U·kg-1组分别与生理盐水组比较,SP含量显著降低(P<0.05).结论:硝酸甘油导致偏头痛症状产生及SP在颈静脉血浆和延髓、脑桥和三叉神经节中的含量增加.局部皮下注射BTX-A可减轻偏头痛大鼠的症状,并能降低颈静脉血浆、脑干和三叉神经节中SP含量,结果提示BTX-A通过抑制"三叉神经血管系统"(trigeminal vascular system,TVS)中SP释放而减轻偏头痛症状.

  8. Inhibitory effect of botulinum toxin type A on isolated aortic contractility-induced by Methoxamine and electric field stimulation%A型肉毒毒素抑制Methoxamine和电场刺激引发主动脉肌条的收缩

    Institute of Scientific and Technical Information of China (English)

    宗晓健; 宋焱峰; 张雪平; 侯一平

    2009-01-01

    目的:观察A型肉毒毒素(botulinum toxin type A,BTX-A)对肾上腺素α受体激动剂Methoxamine (MOA) 和电场刺激(electrical field stimulation,EFS)引发主动脉肌条收缩的作用.方法:在95%O_2,和5%CO_2条件下,将不同条件处理的兔腹主动脉0.5 cm×3.0 cm螺旋形肌条悬挂于37℃ Krebs液的浴槽内,记录收缩张力变化.MOA组:MOA 5 μmol/L引发收缩30 min后分别加入Krebs液(对照,n=10)、BTX-A 50(n=10)或100 U/mL(n=10),30 min后再加入MOA5 μmol/L.EFS组:EFS 50 Hz,80 V,1 ms,60 s持续性引发肌条收缩30 min后分别加入Krebs液(对照,n=10)或BTX-A 50 U/mL(n=10),30 min后再加入MOA 5 μmol/L.结果:BTX-A 50、100 U/mL分别导致MOA诱发的肌条收缩张力下降80%(P<0.01)和95%(P<0.01),再加入MOA引发BTX-A干预过肌条的收缩力仅为对照组的35%(P<0.01)和3%(P<0.01),该抑制作用呈剂量依赖关系.BTX-A 50 U/mL导致EFS诱发的肌条收缩张力下降94%(P<0.01),后续加入MOA引发BTX-A处理过的肌条,收缩力下降为对照组的10%(P<0.01).结论:肾上腺素α受体激动剂MOA和EFS引发兔主动脉肌条收缩,BTX-A抑制MOA和EFS引发动脉平滑肌收缩作用机理与抑制NA的释放和其受体有关.

  9. Effect of Rehabilitation on Long-term Efficacy of Botulinum Toxin Type A on Spastic Cerebral Palsy%康复训练对A型肉毒毒素治疗痉挛型脑瘫长期疗效的影响

    Institute of Scientific and Technical Information of China (English)

    刘建军; 纪树荣; 吴卫红; 张雁; 曾凡勇; 李南玲

    2011-01-01

    Objective To investigate the long-term efficacy of Botulinum toxin type A (BTX-A) on spasticity in cerebral palsy, and the effect of rehabilitation on it. Methods 230 children with spastic cerebral palsy were treated with BTX-A block. The dose of BTX-A was identified with the weight of the children and the Modified Ashworth Scale (MAS). They were divided into group A who exercised more than 2 h/d, and group B who exercised less than 2 h/d or not. They were assessed with Gross Motor Function Measure (GMFM) 1 year after treatment. Results There was no significant difference between group A and B with ages, weight, MAS, GMFM before block (P>0.05), nor with BTX-A effect time (P>0.05). The GMFM improved significantly in both group A and B 1 year after block (P<0.05), and it improved more in group A than group B (P<0.0\\). Conclusion The long-term efficacy of BTX-A block is positive. The rehabilitation training after block can help children to improve their gross motor function.%目的 探讨康复训练对A型肉毒毒素(BTX-A)阻滞术缓解脑瘫患儿痉挛长期疗效的影响。方法 230例痉挛型脑瘫患儿接受BTX-A阻滞术治疗,根据患儿体重和修改的Ashworth量表(MAS)评分确定BTX-A剂量。A组每天进行康复训练2 h以上,B组进行康复训练2 h以下或无训练。1年后评定其粗大运动功能(GMFM)。结果 A、B两组患儿的年龄、体重、术前的MAS、GMFM均无显著性差异(P>0.05),两组患儿肉毒毒素作用时间无显著性差异(P>0.05)。术后1年两组GMFM均较术前提高(P<0.05)。A组GMFM分数提高明显多于B组(P<0.01)。结论 BTX-A阻滞术后有肯定的长期疗效。阻滞术后开展康复训练有助于患儿粗大运动功能的提高。

  10. A型肉毒毒素注射配合康复功能训练对痉挛型偏瘫脑瘫患儿上肢运动功能疗效观察%Effect of Botulinum Toxin Type A Injection Combined with Rehabilitation Functional Training on Upper Extremity Motor Function in Children with Spastic Hemiplegic Cerebral Palsy

    Institute of Scientific and Technical Information of China (English)

    颜华; 张惠佳; 阳伟红; 王益梅; 郭春光; 胡继红; 周平秋; 何金华; 段华林

    2012-01-01

    目的 观察A型肉毒毒素(BTX-A)注射治疗配合康复功能训练对痉挛型偏瘫脑瘫患儿上肢功能障碍的疗效.方法 60例痉挛型偏瘫型脑瘫患儿均接受BTX-A注射治疗,治疗后进行强制性诱导运动训练、物理疗法、肌电生物反馈治疗、作业治疗及家庭训练.在治疗前与治疗3个月后,分别用改良Ashworth痉挛量表(MAS)评定患侧上肢肌张力、关节量角器法测量患侧腕关节主动背伸角度、Peabody运动发育量表(PDMS-II)进行患侧手精细运动商(FMQ)的评估、日常生活活动能力(ADL)量表评估ADL能力以比较观察疗效.结果 治疗后患儿上述指标均较治疗前显著改善(P<0.001).结论 BTX-A注射治疗配合康复功能训练能明显降低痉挛型偏瘫型脑瘫患儿的上肢肌张力,改善关节活动范围,明显提高上肢运动功能.%Objective To observe the effect of Botulinum toxin type A (BTX-A) injection combined with rehabilitation functional training on upper extremity motor function in children with spastic hemiplegic cerebral palsy (CP). Methods 60 spastic hemiplegic CP children were treated with constraint-induced movement therapy (CIMT), physical therapy, electromyographic biofeedback stimulation therapy, occupational therapy, family-based training and so on after injected with BTX-A. The muscle tension of the hemiplegic upper extremity accessed by Modified Ashworth Scale (MAS), the wrist angle of active dorsiextention motion by orthrometer, fine movement quotient (FMQ) by Pea-body developmental motor scale (PDMS- II), and activities of daily living (ADL) were performed to evaluate the effects before and 3 months after treatment. Results These outcomes were improved significantly after treatment (P<0.001). Conclusion BTX-A injection combined with rehabilitation functional training could rapidly reduce spasticity of the upper extremity, increase the range of motion, improve motor function of upper extremity in children with spastic

  11. Effect of botulinum toxin type A on the expression of substance P, calcitionin gene-related peptide,transforming growth factor beta-1 and alpha smooth muscle actin A in wound healing in rats%肉毒毒素A对大鼠皮肤及创面组织中SP、CGRP、TGF-β1和α-SMA的影响

    Institute of Scientific and Technical Information of China (English)

    王琳; 邰宁正; 范志宏

    2009-01-01

    Objective To investigate the effect of botulinum toxin type A on the expression of substance P(SP),calcitionin gene-related peptide(CGRP), transforming growth factor beta-1 (TGF-β1) and alpha smooth muscle actin A(α-SMA) in wound healing. Methods 60 rats were randomly divided into group C (control)、 group L (low-dose) and group H (high-dose), with 20 rats in each group. The wound-healing model was established by excision of four full-thickness skin(1 cm×1 cm, around the injection site) on the back of all SD rats on the 7th day after BTA injection. The wound size was measured and the expression of SP, CGRP, TGF-β1 and α-SMA in wound granulation tissue was assayed by immanohistocbemical staining and computerized image analysis before operation, and 3 days, 7 days and 14 days after operation. Results All the wounds healed 14 days after operation. The wound size in L and H group was not significantly different with that in C group on the 3rd day and 7th day after operation. The positive immuno-staining of SP, CGRP, TGF-β1 and α-SMA in group L and H was significantly weaker than those in C group. Meanwhile, the positive immuno-staining of all above substances in H group was weaker than those in L group significantly. Conclusions Botulinum toxin type A can decrease the expression of SP, CGRP, TGF-β1 and α-SMA in wound healing in a dose-dependent manner with no effect on the healing time.%目的 探讨肉毒毒素A对周围神经末梢释放的感觉神经肽P物质、降钙素基因相关肽及创面组织表达TGF-β1和α-平滑肌肌动蛋白的影响. 方法 60只SD大鼠随机分为空白对照组(C组)、低剂量组(L组)和高剂量组(H组),每组加只大鼠.肉毒毒素A预注射后7 d,在鼠背部建立以4个注射点为中心的创面模型,创面面积1 cm×1 cm.于术前及术后3、7、14 d,测量创面面积,免疫组织化学和Axioplan 2 Imaging图像分析软件检测多聚甲醛灌注后创面P物质、降钙素基因相关肽以及创

  12. Calf muscle volume estimates: Implications for Botulinum toxin treatment?

    DEFF Research Database (Denmark)

    Bandholm, Thomas Quaade; Sonne-Holm, Stig; Thomsen, Carsten;

    2007-01-01

    An optimal botulinum toxin dose may be related to the volume of the targeted muscle. We investigated the suitability of using ultrasound and anthropometry to estimate gastrocnemius and soleus muscle volume. Gastrocnemius and soleus muscle thickness was measured in 11 cadaveric human legs, using...

  13. Evolution of an ancient venom: recognition of a novel family of cnidarian toxins and the common evolutionary origin of sodium and potassium neurotoxins in sea anemone.

    Science.gov (United States)

    Jouiaei, Mahdokht; Sunagar, Kartik; Federman Gross, Aya; Scheib, Holger; Alewood, Paul F; Moran, Yehu; Fry, Bryan G

    2015-06-01

    Despite Cnidaria (sea anemones, corals, jellyfish, and hydroids) being the oldest venomous animal lineage, structure-function relationships, phyletic distributions, and the molecular evolutionary regimes of toxins encoded by these intriguing animals are poorly understood. Hence, we have comprehensively elucidated the phylogenetic and molecular evolutionary histories of pharmacologically characterized cnidarian toxin families, including peptide neurotoxins (voltage-gated Na(+) and K(+) channel-targeting toxins: NaTxs and KTxs, respectively), pore-forming toxins (actinoporins, aerolysin-related toxins, and jellyfish toxins), and the newly discovered small cysteine-rich peptides (SCRiPs). We show that despite long evolutionary histories, most cnidarian toxins remain conserved under the strong influence of negative selection-a finding that is in striking contrast to the rapid evolution of toxin families in evolutionarily younger lineages, such as cone snails and advanced snakes. In contrast to the previous suggestions that implicated SCRiPs in the biomineralization process in corals, we demonstrate that they are potent neurotoxins that are likely involved in the envenoming function, and thus represent the first family of neurotoxins from corals. We also demonstrate the common evolutionary origin of type III KTxs and NaTxs in sea anemones. We show that type III KTxs have evolved from NaTxs under the regime of positive selection, and likely represent a unique evolutionary innovation of the Actinioidea lineage. We report a correlation between the accumulation of episodically adaptive sites and the emergence of novel pharmacological activities in this rapidly evolving neurotoxic clade. PMID:25757852

  14. Botulinum Toxin and Gastrointestinal Tract Disorders: Panacea, Placebo, or Pathway to the Future?

    OpenAIRE

    Lacy, Brian E.; Weiser, Kirsten; Kennedy, Abigail

    2008-01-01

    The history of botulinum toxin is fascinating. First recognized as the cause of botulism nearly 200 years ago, it was originally feared as a deadly poison. Over the last 30 years, however, botulinum toxin has been transformed into a readily available medication used to treat a variety of medical disorders. Interest in the use of botulinum toxin has been particularly strong for patients with spastic smooth muscle disorders of the gastrointestinal tract. Patients with achalasia, diffuse esophag...

  15. Striving for more good days: patient perspectives on botulinum toxin for the treatment of cervical dystonia

    Science.gov (United States)

    Poliziani, Michele; Koch, Marco; Liu, Xierong

    2016-01-01

    Background The recommended reinjection interval for botulinum neurotoxin (BoNT) formulations in the treatment of cervical dystonia (CD) is generally ≥12 weeks, though intervals ≥10 weeks are approved for incobotulinumtoxinA in Europe. However, recurring symptoms can occur before the end of this period. Using qualitative research, we sought a greater understanding of disease burden, unmet patient needs, and barriers to treatment. Methods We conducted online semistructured, focus-group discussions, and online forum follow-up discussions among patients with CD, focusing on disease burden, patient needs, injection cycle preferences, and relationships with health care professionals. A subset of patients was also questioned in telephone interviews about individual experiences of CD and BoNT treatment. All participants were UK residents who had received onabotulinumtoxinA or abobotulinumtoxinA for CD for ≥1 year. Results Thirty-one patients (81% female; mean duration of CD 16.4 [range 4–31] years; mean BoNT injection cycle length 12.8 weeks) participated in the online focus-group and forum follow-up discussions. Of these, seven patients participated in telephone interviews. All had recurring symptoms between treatments, which substantially impacted on their work, family, and social life. Symptom severity fluctuated throughout an injection cycle and differed between patients and across injection cycles. Participants’ relationships with health care professionals and treatment satisfaction varied greatly. Many participants wanted longer-lasting and/or more stable symptom relief with shorter and/or more flexible injection intervals, according to individual needs. Lack of health care resources, long journeys to treatment centers, and immunogenicity/side-effect concerns were perceived as the main barriers to more flexible treatment. Conclusion The high burden of recurring primary and secondary symptoms of CD considerably affects patients’ quality of life. Patient

  16. Effect of botulinum toxin type A injection on hypertrophic scar in rabbit ear model%A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

    Institute of Scientific and Technical Information of China (English)

    王琳; 邰宁正; 范志宏

    2009-01-01

    目的 研究A型肉毒毒素对兔耳增生性瘢痕组织的影响.方法 8只日本大耳白兔,体重3 kg,建立兔耳增生性瘢痕模型.将兔耳创面分为A型肉毒毒素治疗组(T组)和瘢痕组(S组),每组48个创面.大体观察创面愈合时间和瘢痕增生情况.术后28 d,同法另取4只兔子的兔耳腹面健康皮肤为空白组(B组),收集标本.测量S、T组标本HE切片的瘢痕增生指数HI,流式细胞仪分析2组标本中成纤维细胞的细胞周期,western-blot检测S、T、B组标本中Ⅰ、Ⅲ型胶原的蛋白表达.结果 ①T组标本的瘢痕增生指数HI较S组显著降低,P<0.01;②蛋白水平上,T组的胶原Ⅰ、Ⅲ蛋白表达和胶原Ⅰ/Ⅲ比值均较s组显著降低,P<0.01;③S组分布于G2-M期和S期的成纤维细胞较T组显著增多,而静止期G0-G1的细胞则显著减少,P<0.05.结论 A型肉毒毒素局部应用能抑制兔耳增生瘢痕的形成.抑制成纤维细胞的增殖活性,减少瘢痕组织中Ⅰ、Ⅲ型胶原的合成,降低胶原Ⅰ/Ⅲ比值,为其治疗增生性瘢痕的临床应用提供了一定的理论依据.%Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more

  17. Anti-inflammatory effects of botulinum toxin type A in a rat model of arthritis%A型肉毒毒素关节腔注射对佐剂性关节炎大鼠的抗炎效果观察

    Institute of Scientific and Technical Information of China (English)

    王开乐; 初晓; 张仲义; 王琳; 沈娜娜; 薛俊强; 李铁山

    2016-01-01

    Objective To verify the anti-inflammatory effects of intra-articular injection of botulinum toxin type A (BoNT/A) on adjuvant-induced arthritis using a rat model.Methods A murine model of chronic ankle arthritis was established in 90 Wistar rats by injection of 0.1 ml of complete Freund adjuvant (CFA) into the pads of their left paws.They were then randomly divided into a BoNT group (n =30) which received an intra-articular injection of 0.1 ml (20 IU) of BoNT/A,an NS group (n=30) which received intra-articular injection of0.1 ml of normal saline solution and a sham group (n =30) which were punctured without any injection.In addition,30 normal rats formed a control group.Infrared thermal imaging was performed and an index of arthritis was evaluated every three days.The infrared thermal imaging revealed the expression of interleukin-1β (IL-1β) through hematoxy-eosin (HE) staining.Results The arthritis index began to increase 3 days after the injection of CFA and it had increased significantly after 10 days,reaching a peak value of 18,24 days after the injection.The infrared thermal imaging showed that the temperature in the right paw increased greatly after the injection.Following the development of arthritis,the temperature declined gradually,arriving at a steady temperature of between 37.5 and 38.0 ℃ in both ankles 20 days after the injection.The average temperature in both paws of the BoNT group had decreased significantly more by 7 and 14 days after the injection than in the NS and sham groups.The expression of IL-1β in the synovium of the ankle joint also had decreased significantly more in the BoNT group after 7 and 14 days.HE scoring showed an obvious histopathologic change in the hypertrophic synovium,inflamnatory cell infiltration,cartilage destruction and exposure of subchondral bone after 7 and 14 days compared with right after the injection in all groups except the control group.Moreover,the average HE scores of the BoNT group rats after 7 and 14

  18. Innovative mode of action based in vitro assays for detection of marine neurotoxins

    OpenAIRE

    Nicolas, J.A.Y.

    2015-01-01

    Innovative mode of action based in vitro assays for detection of marine neurotoxins J. Nicolas, P.J.M. Hendriksen, T.F.H. Bovee, I.M.C.M. Rietjens Marine biotoxins are naturally occurring compounds produced by particular phytoplankton species. These toxins often accumulate in seafood and thereby represent a threat to consumers. Regulatory limits have been set for lipophilic marine biotoxins (diarrhetic shellfish poisons (DSPs) and azaspiracids (AZPs)) and for most marine neurotoxins (amnesic ...

  19. Botulinum Toxin for the Treatment of Tremor and Tics.

    Science.gov (United States)

    Lotia, Mitesh; Jankovic, Joseph

    2016-02-01

    The therapeutic applications of botulinum toxin (BoNT) have grown manifold since its initial approval in 1989 by the U.S. Food and Drug Administration for the treatment of strabismus, blepharospasm, and other facial spasms. Although it is the most potent biologic toxin known to man, long-term studies have established its safety in the treatment of a variety of neurologic and nonneurologic disorders. Despite a paucity of randomized controlled trials, BoNT has been found to be beneficial in treating a variety of tremors and tics when used by clinicians skilled in the administration of the drug for these hyperkinetic movement disorders. Botulinum toxin injections can provide meaningful improvement in patients with localized tremors and tics; in some cases, they may be an alternative to other treatments with more undesirable adverse effects.

  20. Intraoral administration of botulinum toxin for trigeminal neuropathic pain.

    Science.gov (United States)

    Herrero Babiloni, Alberto; Kapos, Flavia P; Nixdorf, Donald R

    2016-06-01

    This article presents 2 cases of different neuropathic trigeminal pain conditions treated with intraoral botulinum toxin injections. There is a growing body of evidence to support the use of this substance when administered subcutaneously in the treatment of neuropathic pain, such as in extraoral injections for trigeminal neuralgia. However, reports of intraoral submucosal administration are still lacking. In the 2 cases presented here, neuropathic pain was refractory to treatment with an important intraoral peripheral component, so onabotulinum toxin A was introduced as an adjuvant therapy. The technique, doses, and dilution are discussed. The patients reported significant reductions in pain frequency and intensity, with minimal side effects of temporary mucosal dryness and smile droopiness. The analgesic benefits of botulinum toxin may be utilized to address intraoral neuropathic pain. Further studies are needed to confirm safety and effectiveness in larger samples. PMID:27181448

  1. Inhibition of Clostridium botulinum by antioxidants, phenols, and related compounds.

    OpenAIRE

    Reddy, N R; Pierson, M. D.; Lechowich, R. V.

    1982-01-01

    A total of 75 compounds, including antioxidants, preservatives, gallic acid and p-hydroxybenzoic acid esters, hydroquinones, hydroxyquinolines, phenol derivatives, and related compounds, were screened for their antibotulinal activity in prereduced Thiotone-yeast extract-glucose broth. The most effective inhibitors of Clostridium botulinum growth and toxin production were long-chain esters of p-hydroxybenzoic acid and gallic acid, antioxidants, and butylphenol derivatives. The antioxidant nord...

  2. Botulinum toxin industry : is it a profitable industry to enter?

    OpenAIRE

    Salopek, Barbara

    2009-01-01

    This paper examines the botulinum toxin industry and is it profitable to enter. In particular, the paper aims to define whether the industry structure is attractive because an industry is considered profitable only if it has attractive structure. This has been explored through applying the Porter’s five forces framework which provides a good understanding of farces shaping the industry and thus help to define the industry structure. It has been shown that the BTX industry has b...

  3. Botulinum toxin for treating muscular temporomandibular disorders: a systematic review

    OpenAIRE

    Eduardo Machado; Lívia Zuchetto dos Santos; Lilian Gonçalves Custódio; Paulo Afonso Cunali

    2012-01-01

    OBJECTIVE: This study, through a systematic literature review, aims to analyze the effectiveness of Botulinum Toxin as a treatment for masticatory myofascial pain and muscles temporomandibular disorders (TMD). METHODS: Survey in research bases: MEDLINE, Cochrane, EMBASE, Pubmed, Lilacs and BBO, between the years of 1966 and April 2011, with focus in randomized or quasi-randomized controlled clinical trials, blind or double-blind. RESULTS: After applying the inclusion criteria, 4 articles comp...

  4. C型肉毒梭菌肉毒毒素的提取与鉴定%Extraction and Identification of Botulinum Toxins

    Institute of Scientific and Technical Information of China (English)

    薛正坤; 吴伟; 张晶; 李超毅; 吴辉; 李莲瑞

    2013-01-01

    [目的]通过对C型肉毒梭菌肉毒毒素的提取与鉴定,为类毒素和抗毒素的制备及抗原性分析奠定基础.[方法]将分离鉴定得到的C型肉毒梭菌通过扩大培养、产毒培养后将产生的肉毒毒素采用除菌过滤、硫酸铵盐盐析、离心、透析、浓缩的方法从产毒培养基中分离提取出来.再将提取的肉毒毒素通过SDS-PAGE鉴定毒素蛋白的分子量.[结果]分离出来的毒素蛋白重链和轻链分别在98和53 KDa左右,与C型肉毒毒素的理论分子量相符.[结论]提取的肉毒毒素是C型肉毒毒素.%[Objective] The project aims to lay a foundation for the preparation and antigenicity analysis of the toxoid and antitoxic through extraction and identification of the type C clostridium botulinumn. [Method] The separated and identified type C clostridium botulinum underwent expanding training, and after that, the procreant poison produced botulinum toxin through such methods as the aseptic filtration, ammonium sulfate salting separation, centrifugal treatment, dialysis and enrichment processes. Then the extracted toxin went through SDS - PAGE for its toxin protein molecular weight to be identified. [ Result ] From the experiments, the abstracted toxin protein heavy chain was about 98 KDA and light chain about 53 KDA, which was consistent with molecular weight of type C botulinum toxin theory. [ Conclusion] What we have extracted is C - type Clostridium botulinum.

  5. Efficacy of intraarticular botulinum toxin A and intraarticular hyaluronate plus rehabilitation exercise in patients with unilateral ankle osteoarthritis: a randomized controlled trial

    OpenAIRE

    Sun, Shu-Fen; Hsu, Chien-Wei; Lin, Huey-Shyan; Chou, Yi-Jiun; Chen, Jun-yang; Wang, Jue-Long

    2014-01-01

    Background There was an increasing requirement for novel treatments of osteoarthritis (OA). The aim was to compare the efficacy of intraarticular Botulinum toxin type A (BoNT-A) and intraarticular hyaluronate plus rehabilitation exercise in patients with ankle OA. Methods This was a prospective, randomized, assessor-blinded study with a 6-month follow-up period, conducted in the outpatient rehabilitation department at a university-affiliated tertiary care medical center. Seventy-five patients...

  6. Quantitative evaluation for spasticity of calf muscle after botulinum toxin injection in patients with cerebral palsy: a pilot study

    OpenAIRE

    Lin, Yu-Ching; Lin, I-Ling; Chou, Te-Feng Arthur; Lee, Hsin-Min

    2016-01-01

    Background Cerebral palsy (CP) is the most common pediatric disease to cause motor disability. Two common symptoms in CP are spasticity and contracture. If this occurred in the ankle plantar flexors of children with CP, it will impair their gait and active daily living profoundly. Most children with CP receive botulinum toxin type A (BoNT-A) injection to reduce muscle tone, but a knowledge gap exists in the understanding of changes of neural and non-neural components of spasticity after injec...

  7. Botulinum toxin A in the treatment of anal fissure

    Directory of Open Access Journals (Sweden)

    Stanković Nebojša

    2004-01-01

    Full Text Available Background. This paper presents our first experience in the treatment of primary anal fissure by injection of botulinum A toxin into the internal sphincter. Methods. The study group of the retrospective study included 12 outpatients (8 females and 4 males, mean age 42 (range 18-60. During the period 2000-2003, after unsuccessful conservative treatment, patients were treated with the injections of botulinum A toxin, 100 units on both sides of the anal fissure laterally into the internal anal sphincter (50 units on either side. The patients were clinically evaluated 3, 7, and 30 days, and 3 and 6 months after the treatment. Results. Three fissures had healed after a month, and seven after 3 months. Two remained unhealed, but asymptomatic. There was no incontinence of flatus or faeces after 3 months of the treatment. After temporary healing, two fissures relapsed after 6 months, and these patients had the adequate tonus of the anal sphincter muscles. Except for the temporary incontinence, there were neither other side-efects, nor serious complications. Conclusion. Injection of botulinum A toxin achieved good results in the treatment of anal fissure. The appropriate use makes this method safe as an alternative to surgical treatment of anal fissure.

  8. A Beautician's Dystonia: Long-Lasting Effect of Botulinum Toxin

    Science.gov (United States)

    Di Martino, Siria; Dalise, Stefania; Lamola, Giuseppe; Venturi, Martina; Rossi, Bruno; Chisari, Carmelo

    2014-01-01

    Treatment options for dystonia are not curative but symptomatic; the treatment of choice for focal dystonias is repeated botulinum toxin injections. Here, we present the case of a 46-year-old beautician with focal dystonia in her left hand that affected her ability to work. Pharmacological treatment with clonazepam and gabapentin failed to resolve her symptoms and was discontinued due to side effects (sleepiness, gastrointestinal disorders). Intramuscular injection of botulinum toxin (incobotulinumtoxinA, Xeomin) into the extensor digitorum communis (35 U), flexor carpi radialis (35 U), and flexor digitorum superficialis (30 U) muscles resulted in complete resolution of symptoms at clinical assessments at 1, 3, 6, and 10 months after the injections, confirmed by the results of surface electromyography 10 months after treatment. The patient was able to work again 1 month after treatment. No reinjection has been necessary at the last evaluation (12 months after treatment). In conclusion, botulinum toxin is an effective treatment for focal dystonia that can have long-lasting effects and can improve patients' ability to work and quality of life. PMID:25143844

  9. Optimisation of protocol for Clostridium botulinum detection in mink feed

    Directory of Open Access Journals (Sweden)

    Grenda Tomasz

    2015-09-01

    Full Text Available As the test material mink feed with natural microflora was used. The analyses were conducted using Wrzosek and TPGY broth media, and Willis–Hobbs and Zeissler differential agar media. Wrzosek, Willis–Hobbs, and Zeissler media are described in Polish Standards approved by the National Standards Body in Poland and routinely used in detection of anaerobic bacteria in Poland. Detection and identification of C. botulinum was performed with a previously validated real-time PCR method based on ntnh gene detection, which is common in all C. botulinum toxotypes. The use of Wrzosek broth and Zeissler agar in routine analyses for detection and identification of C. botulinum was ineffective and limited. The obtained results showed the highest culturing process effectiveness in TPGY broth with 72 h incubation at 30°C and isolation on Willis–Hobbs agar. The real-time PCR method based on ntnh gene detection used in this study could be utilised as a supplementary tool to the mouse lethality assay.

  10. Studies on the irradiation of toxins of Clostridium botulinum and Staphylococcus aureus

    Energy Technology Data Exchange (ETDEWEB)

    Rose, S.A.; Bailey, N.E.; Stringer, M.F. (Campden Food and Drink Research Association, Chipping Campden (UK)); Modi, N.K.; Tranter, H.S. (Centre for Applied Micobiological Research, Porton Down, (UK)); Hambleton, P. (Porton International plc, London (UK))

    1989-09-01

    The effects of irradiation of Clostridium botulinum neutotoxin type A (BNTA) and staphylococcal enterotoxin A (SEA) in gelatin phosphate buffer and cooked mince beef slurries were investigated. Estimation of toxins by immunoassays showed that in buffer, toxins were destroyed by irradiation at 8.0 kGy; in mince slurries however, 45% of BTNA and 27-34% of SEA remained after this level of irradiation. At 23.7 kGy, over twice the dose of irradiation proposed for legal acceptance in the UK, 15% of BNTA and 16-26% of SEA still remained. Increasing concentrations of mince conferred increased protection against the effect of irradiation on both toxins. The biological activity of BNTA was more sensitive to irradiation than the immunological activity. Staphylococcal enterotoxin was more resistant to irradiation than BNTA. Irradiation should therefore only be used in conjunction with good manufacturing practices to prevent microbial proliferation and toxin production prior to irradiation. (author).

  11. Muscle selection for treatment of cervical dystonia with botulinum toxin : A systematic review

    NARCIS (Netherlands)

    Nijmeijer, S. W. R.; Koelman, J. H. T. M.; Kamphuis, D. J.; Tijssen, M. A. J.

    2012-01-01

    Rationale: Cervical dystonia, also called spasmodic torticollis, is the most common form of (primary) dystonia. Intramuscular injections with botulinum toxin are the first line of treatment for cervical dystonia. To optimise the treatment response to botulinum toxin correct muscles should be selecte

  12. Risk assessment of proteolytic Clostridium botulinum in canned foie gras.

    Science.gov (United States)

    Membré, Jeanne-Marie; Diao, Moctar; Thorin, Chantal; Cordier, Grégoire; Zuber, François; André, Stéphane

    2015-10-01

    In this study, a risk assessment of proteolytic Clostridium botulinum in canned foie gras was performed, the number of illnesses per year in France due to C. botulinum in foie gras was estimated. Data on initial level in raw materials were collected at manufacturers and analysed using a Negative Binomial distribution. The effect of the usual foie gras heat treatment (equivalent time at 121 °C: F0=0.5 min) was considered at two levels: first, it led to an inactivation (estimated to 2.3 log); second it led to a spore injury and then to a spore inhibition. This latter effect was assessed by analysing data from a challenge test study carried out with Clostridium sporogenes spores in the foie gras product. The probability of spore recovering after thermal inhibition was estimated to 9.5×10(-8) (corresponding to 7.0 log). The data on the consumption pattern were collected on the French market. The Quantitative Microbiological Risk Assessment (QMRA) model and all the assumptions are reported in detail in the study. The initial contamination of raw materials, effect of thermal treatment on microbial inactivation and spore inhibition were handled mathematically using a probabilistic framework, considering only the variability dimension. The model was implemented in Excel and run through Monte Carlo simulation, using @Risk software. In parallel, epidemiological data collected from the French Institute for Public Health Surveillance during the period 2001-2012 were used to estimate an Appropriate Level Of Protection (ALOP) and then a Food Safety Objective (FSO): ALOP equalled to 2.5×10(-3) illnesses per million inhabitant per year, FSO equalled to 1.4×10(-9) foie gras portions containing C. botulinum spore (expressed in decimal logarithm, FSO=-8.9). The QMRA model output values were smaller, but on the same order of magnitude as these two figures: 8.0×10(-4) illnesses per million inhabitants per year, and, 4.5×10(-10) (-9.3 log) foie gras portions containing C

  13. Risk assessment of proteolytic Clostridium botulinum in canned foie gras.

    Science.gov (United States)

    Membré, Jeanne-Marie; Diao, Moctar; Thorin, Chantal; Cordier, Grégoire; Zuber, François; André, Stéphane

    2015-10-01

    In this study, a risk assessment of proteolytic Clostridium botulinum in canned foie gras was performed, the number of illnesses per year in France due to C. botulinum in foie gras was estimated. Data on initial level in raw materials were collected at manufacturers and analysed using a Negative Binomial distribution. The effect of the usual foie gras heat treatment (equivalent time at 121 °C: F0=0.5 min) was considered at two levels: first, it led to an inactivation (estimated to 2.3 log); second it led to a spore injury and then to a spore inhibition. This latter effect was assessed by analysing data from a challenge test study carried out with Clostridium sporogenes spores in the foie gras product. The probability of spore recovering after thermal inhibition was estimated to 9.5×10(-8) (corresponding to 7.0 log). The data on the consumption pattern were collected on the French market. The Quantitative Microbiological Risk Assessment (QMRA) model and all the assumptions are reported in detail in the study. The initial contamination of raw materials, effect of thermal treatment on microbial inactivation and spore inhibition were handled mathematically using a probabilistic framework, considering only the variability dimension. The model was implemented in Excel and run through Monte Carlo simulation, using @Risk software. In parallel, epidemiological data collected from the French Institute for Public Health Surveillance during the period 2001-2012 were used to estimate an Appropriate Level Of Protection (ALOP) and then a Food Safety Objective (FSO): ALOP equalled to 2.5×10(-3) illnesses per million inhabitant per year, FSO equalled to 1.4×10(-9) foie gras portions containing C. botulinum spore (expressed in decimal logarithm, FSO=-8.9). The QMRA model output values were smaller, but on the same order of magnitude as these two figures: 8.0×10(-4) illnesses per million inhabitants per year, and, 4.5×10(-10) (-9.3 log) foie gras portions containing C

  14. A型肉毒素治疗脑卒中康复患者肢体痉挛的护理体会%The Nursing Experience of Botulinum Toxin A in the Treatment of Limb Spasm for Stroke Rehabilitation Patients

    Institute of Scientific and Technical Information of China (English)

    王明霞; 尚小侠; 史盼

    2016-01-01

    目的:探讨A型肉毒素注射治疗肢体痉挛的护理。方法对我科31例脑卒中后肢体痉挛患者进行A型肉毒素注射治疗的临床护理进行回顾性分析。结果31例脑卒中后肢体痉挛患者经A型肉毒素注射治疗,配合适时护理,治疗后均获得良好效果,无严重并发症。结论对患者规范细致周到的护理有助于提高肉毒素注射疗效,减少并发症。%Objective To explore Nursing care of patients with limb spasm treated with botulinum toxin type A injection.Methods The clinical nursing of 31 patients with limb spasm after stroke in our department were retrospectively analyzed with the clinical nursing of A type botulinum toxin injection. Results31 cases of cerebral apoplexy patients with limb spasm were treated with botulinum toxin type A,with timely care,good results were obtained after treatment,no serious complications.Conclusion Careful and meticulous care of patients is helpful to impove the efficacy of botulinum toxin injection and reduce the complications.

  15. Botulinum toxin - neuropathic pain: Safety and efficacy of repeated injections of botulinum toxin A in peripheral neuropathic pain (BOTNEP): a randomised, double-blind, placebo-controlled trial.

    Science.gov (United States)

    2016-08-12

    When treated with botulinum toxin A, those patients with peripheral neuropathic pain and allodynia (triggering of pain from stimuli which do not normally provoke pain) at baseline, would appear to have a better outcome. PMID:27514345

  16. A型肉毒毒素在斜视治疗中的应用%Botulinum toxin A in treatmen of the strabismus

    Institute of Scientific and Technical Information of China (English)

    王贝贝; 胡依博; 王素萍(综述); 王瑞峰(审校)

    2016-01-01

    Botulinum toxin A injections the extraocular muscles which can cause temporary paralysis to change the positiong of eye in order to treatment of strabismus.And it can produce the extraocular muscle temporary weakness and permanent eye position change, with the advantages of repeated injections, simple operation and less side effects, etc. In this paper, botulinum toxin type A application in strabismus treatment were summarized.%肉毒毒素A型注射眼外肌可造成暂时性麻痹来改变眼位治疗斜视,可产生眼外肌暂时的无力和永久的眼位改变,并具有可重复注射,操作简单和副作用少等优点。本文对肉毒毒素A型在斜视治疗中的应用进行综述。

  17. Effect of botulinum toxin type A injection on the expression of substance P, TGF-β1 and α-SMA in rabbit ear model of hypertrophic scar%A型肉毒毒素对兔耳增生性瘢痕组织中P物质、β1转移生长因子、α平滑肌肌动蛋白的影响

    Institute of Scientific and Technical Information of China (English)

    王琳; 邰宁正; 范志宏

    2009-01-01

    Objective To investigate the effect of botulinum toxin type A injection on the expres-sion of substance P, TGF-β1 and α-SMA in rabbit ear model of hypertrophic scar. Methods The hyper-trophic sear model was established in 24 Japanese rabbits'ears. The wounds in ventral surface of ear were divided in Group I (lateral wounds) and Group S (medial wounds), 3 wounds each side per ear, totally 72 wounds each group. The wound-healing time and the growth of scar were observed and recorded. On post-wounding day 28, the wounds were created in another 6 rabbits in the same way and the normal skin were harvested as Group C. Likewise, the scar samples in Groups I and S were harvested. The mRNA expression of substance P, TGF-β1 and α-SMA were detected quantitatively by using real-time PCR and α-SMA was also detected with Western blot. Results No difference between the ratio of healed or infec-tious wounds on post-wounding day 14. The mRNA expression of SP, TGF-β1 and α-SMA in Group I was significantly lower than Group S, but higher than those in Group C (PI组>C组.结论 BTA注射不延迟创面愈合,并减少了兔耳增生瘢痕中SP、TGF-β1和α-SMA的mRNA表达,为其治疗增生性瘢痕的临床应用提供了一定的理论依据.

  18. Use of botulinum toxin-A for musculoskeletal pain in patients with whiplash associated disorders [ISRCTN68653575

    Directory of Open Access Journals (Sweden)

    Juan Francisco J

    2004-02-01

    Full Text Available Abstract Background Whiplash associated disorder is commonly linked to motor vehicle accidents and sports injuries. Cervical injury is attributed to rapid extension followed by neck flexion. The exact pathophysiology of whiplash is uncertain but probably involves some degree of aberrant muscle spasms and may produce a wide range of symptoms. The most commonly prescribed pharmacological agents for initial treatment of whiplash-associated pain are oral muscle relaxants and nonsteroidal anti-inflammatory drugs. However, potential systemic adverse effects limit these agents. Physical interventions such as mobilization, manipulation, and exercises have proved beneficial for pain and dysfunction but only on a time-limited basis. Little evidence suggests that physical therapy specifically aimed at the musculature (e.g., transcutaneous electrical nerve stimulation, ultrasonography, heat, ice, and acupuncture improves prognosis in acute whiplash associated disorder. A new approach to treatment is the use of botulinum toxin, which acts to reduce muscle spasms. Methods/design This is a prospective, randomized, controlled clinical trial and botulinum toxin-A (Botox® injections will be compared with placebo injections. The primary objective is to determine the efficacy of Botox® in the management of musculoskeletal pain in whiplash associated disorders. Discussion Botulinum toxin type-A toxin has been studied in small trials on whiplash associated disorder patients and has generally been found to relieve pain and improve range of motion. Specifically, we seek to assess the efficacy of Botox® in reducing pain and to improve the cervical spine range of movement, during the 6-month trial period.

  19. Esporos de Clostridium botulinum em mel comercializado no Estado de São Paulo e em outros Estados brasileiros Clostridium botulinum spores in honey commercialized in São Paulo and other Brazilian states

    Directory of Open Access Journals (Sweden)

    Adriana Valim Ferreira Ragazani

    2008-04-01

    Full Text Available O botulismo infantil tem afetado crianças abaixo de um ano de idade em várias regiões do mundo, e o mel tem sido identificado como uma das mais importantes fontes de intoxicação alimentar. Apesar disso, há dados escassos sobre o botulismo entre crianças no Brasil, especialmente no tipo de alimento comercial mais implicado nesta patologia. Este estudo pretendeu investigar a presença de esporos de Clostridium botulinum em mel comercializado no Brasil. Cem amostras de mel comercializado em seis diferentes Estados brasileiros (SP, MG, GO, CE, MT, SC foram pesquisados para a presença de esporos de Clostridium botulinum, usando o choque térmico, seguido pela inoculação em caldo Cooked Meat Medium (Difco® e incubado em condições anaeróbias. As culturas positivas foram analisadas através de esfregaços corados pelo Gram e semeadas em placas de Reinforced Clostrideo Agar (Difco® e placas de Sulfito Polimixina Sulfadiazina -SPS (Difco®, as quais foram incubadas em condições anaeróbicas para obter colônias desta bactéria. As colônias positivas foram submetidas a teste de toxicidade através da inoculação em camundongos susceptíveis e caracterização bioquímica. Foram encontradas colônias de Clostridium botulinum que produzem toxinas ativas em 7% das amostras de mel comercial, realçando a relevância deste microrganismo para a saúde pública devido ao alto risco potencial de o mel comercializado nestas regiões brasileiras causar o botulismo infantil, especialmente em crianças abaixo de um ano de idade.Infant botulism has been affecting children under one year of age in several regions of the world and honey has been identified as one of the most important source of this food borne disease. Despite this, there are scarce data about botulism among children in Brazil, specially the type commercial food most implicated in this pathology. This study aimed at investigating the presence of spores of Clostridium botulinum in honey

  20. Abnormal motor cortex excitability during linguistic tasks in adductor-type spasmodic dysphonia.

    Science.gov (United States)

    Suppa, A; Marsili, L; Giovannelli, F; Di Stasio, F; Rocchi, L; Upadhyay, N; Ruoppolo, G; Cincotta, M; Berardelli, A

    2015-08-01

    In healthy subjects (HS), transcranial magnetic stimulation (TMS) applied during 'linguistic' tasks discloses excitability changes in the dominant hemisphere primary motor cortex (M1). We investigated 'linguistic' task-related cortical excitability modulation in patients with adductor-type spasmodic dysphonia (ASD), a speech-related focal dystonia. We studied 10 ASD patients and 10 HS. Speech examination included voice cepstral analysis. We investigated the dominant/non-dominant M1 excitability at baseline, during 'linguistic' (reading aloud/silent reading/producing simple phonation) and 'non-linguistic' tasks (looking at non-letter strings/producing oral movements). Motor evoked potentials (MEPs) were recorded from the contralateral hand muscles. We measured the cortical silent period (CSP) length and tested MEPs in HS and patients performing the 'linguistic' tasks with different voice intensities. We also examined MEPs in HS and ASD during hand-related 'action-verb' observation. Patients were studied under and not-under botulinum neurotoxin-type A (BoNT-A). In HS, TMS over the dominant M1 elicited larger MEPs during 'reading aloud' than during the other 'linguistic'/'non-linguistic' tasks. Conversely, in ASD, TMS over the dominant M1 elicited increased-amplitude MEPs during 'reading aloud' and 'syllabic phonation' tasks. CSP length was shorter in ASD than in HS and remained unchanged in both groups performing 'linguistic'/'non-linguistic' tasks. In HS and ASD, 'linguistic' task-related excitability changes were present regardless of the different voice intensities. During hand-related 'action-verb' observation, MEPs decreased in HS, whereas in ASD they increased. In ASD, BoNT-A improved speech, as demonstrated by cepstral analysis and restored the TMS abnormalities. ASD reflects dominant hemisphere excitability changes related to 'linguistic' tasks; BoNT-A returns these excitability changes to normal.

  1. A型肉毒素抑制电场刺激及乙酰胆碱引发的大鼠胃体胃底离体平滑肌收缩%Inhibitory Effect of Botulinum Toxin Type A on Gastric Body and Gastric Fundus Smooth Muscle Contractility Induced by EFS and ACh-induced in Rats in vitro

    Institute of Scientific and Technical Information of China (English)

    周媛媛; 李超彦; 侯一平

    2012-01-01

    Objective: To observe the effect of botulinum toxin type A ( BTX-A) , electrical field stimulation (EFS) and acetylcholine ( ACh) on spontaneous contractility in gastric body and gastric fundus smooth muscle. Method; Muscle strips in gastric body and gastric fundus were prepared, and subdivided randomly into control group, EFS group, BTX-A (10 U -mL-1) group, BTX-A (10 U -mL-1) + EFS group, ACh (100 Ixmol-L-1) group, ACh (100 (xmol-L-1) + BTX-A (10 U -mL-1) group, Ach (100 μmol - L-1) + Atropine (1 (μ.mol -L-1) group. The data were recorded by physiological experimental system of BL-420. Result: EFS enhanced the tension ( P < 0. 05 ) and amplitude ( P < 0. 01 ) in gastric body contractility, and similar results was observed in gastric fundus contractility; BTX-A decreased spontaneous contractile tension and amplitude (P <0. 01) in gastric body and tension ( P < 0. 05 ) in gastric fundus; BTX-A inhibited EFS-induced smooth muscle contractility including tension and amplitude ( P < 0. 01 ) in gastric body, tension (P < 0. 01 ) and amplitude ( P < 0. 05 ) in gastric fundus, BTX-A inhibited ACh-induced smooth muscle contractility including tension and amplitude (P <0. 01) in gastric body and gastric fundus. Conclusion; EFS enhances smooth muscle spontaneous contractility in gastric body and gastric fundus; BTX-A inhibits gastric body and gastric fundus smooth muscle spontaneous contractility; BTX-A inhibits EFS and ACh-induced smooth muscle contractility in gastric body and gastric fundus.%目的:观察大鼠胃体、胃底离体平滑肌条自发性收缩及电场刺激( EFS)、乙酰胆碱(ACh)和A型肉毒素( BTX-A)对肌条收缩的影响,并探讨其机制.方法:取大鼠胃体胃底平滑肌制备肌条,肌条随机分为对照组、EFS组、BTX-A(10 U·mL-1)组、BTX-A(10 U·mL-1)+ EFS组、ACh(100 μmol· L-1)组、ACh(100 μmol·L-1)+BTX-A(10 U·mL-1)组、ACh(100 μmol· L-1)+阿托品(1 μmol· L-)组,采用Biolap 420E生物机能实验系统记录肌

  2. 踝足矫形器联合肉毒毒素治疗痉挛型脑性瘫痪儿童尖足畸形的疗效观察%Ankle-foot orthosis combined with botulinum toxin type A injection in treating tip foot deformity in children with cerebral palsy

    Institute of Scientific and Technical Information of China (English)

    周陶成; 童光磊; 张敏; 李司南; 易昕; 陈露露; 温祖芳; 康倩倩; 陈婧

    2015-01-01

    目的 观察踝足矫形器(A FO)联合A型肉毒毒素(BTX-A)肌肉注射治疗痉挛型脑瘫患儿尖足的疗效.方法 痉挛型脑瘫患儿尖足畸形50例,采用随机数字表法将其分为对照组和观察组,每组25例,对照组患儿采用BTX-A肌肉注射与运动康复训练,观察组在对照组基础上加用AFO训练,与其运动康复训练同步.分别于治疗前、治疗后1、3和6个月,使用量角器测量踝关节的被动背屈角(APROM),改良Ashworth量表的评分(MAS)以及粗大运动功能量表(GMFM-88)的D区(站立)和E区(走跑跳)评分.结果 治疗前,观察组APROM、MAS和GMFM(D、E)分别为(109.25±12.38)°、(3.12±0.56)分和(55.32±11.23)分,与对照组[(107.11±13.44)°、(3.05±0.66)分和(56.21±10.81)分]比较,差异均无统计学意义(P>0.05).2组患儿的APROM、MAS和GMFM评分在治疗后1、3和6个月时与组内与治疗前比较,差异均有统计学意义(P<0.05);治疗后,观察组APROM、MAS和GMFM评分分别与对照组治疗比较,差异亦均有统计学意义(P<0.05).2组治疗后各时间点之间的评估指标比较,观察组差异均有统计学意义;对照组治疗后1个月与治疗后3个月比较,差异有统计学意义(P<0.05),治疗后3个月和治疗后6个月比较,差异无统计学意义(P>0.05).结论 AFO联合BTX-A疗法能进一步缓解痉挛型脑瘫患儿的下肢痉挛程度,减轻尖足异常姿势,维持时间更持久.%Objective To observe the clinical efficacy of ankle-foot orthosis (AFO) combined with botulinum toxin type A (BTX-A) injection in treating tip foot deformity in children with cerebral palsy (CP).Methods Fifty CP children with tip foot deformity were selected and randomly divided into a control group and an observation group according to a random number table.Both groups were given BTX-A injection, and the observation group was additionally treated with AFO.All children were assessed before, 1 month, 3 months and 6 months after the treatment

  3. An Overview of Type E Botulism in China

    Institute of Scientific and Technical Information of China (English)

    SI-WU FU; CHEN-HUAI WANG

    2008-01-01

    The geographical distribution of C. botulinum type E and its associated disease, type E botulism in China, is different from that in other areas of the world. Cases of type E botulism generally arise in costal regions. In China, however, type E botulism is found primarily in the Qinghai-Tibet plateau of northwest China far from the ocean, at an altitude of approximately 4-5 kin. The foods most commonly associated with the disease are fermented grain and beans as well as raw meat. A suspected outbreak of type E botulism poisoning in the central costal region of China in the 1990s prompted the collection and analysis of samples of mud, sand, and fish from the region. The toxin produced by type E botulinum was found in these samples. Surprisingly, though, upon further analysis, the strain isolated from the samples was identified not as type E C. botulinum, but as the neurotoxigenic bacterium Clostridium butyricum.

  4. In Silico Analysis for the Study of Botulinum Toxin Structure

    Science.gov (United States)

    Suzuki, Tomonori; Miyazaki, Satoru

    2010-01-01

    Protein-protein interactions play many important roles in biological function. Knowledge of protein-protein complex structure is required for understanding the function. The determination of protein-protein complex structure by experimental studies remains difficult, therefore computational prediction of protein structures by structure modeling and docking studies is valuable method. In addition, MD simulation is also one of the most popular methods for protein structure modeling and characteristics. Here, we attempt to predict protein-protein complex structure and property using some of bioinformatic methods, and we focus botulinum toxin complex as target structure.

  5. Advances in Assays and Analytical Approaches for Botulinum Toxin Detection

    Energy Technology Data Exchange (ETDEWEB)

    Grate, Jay W.; Ozanich, Richard M.; Warner, Marvin G.; Bruckner-Lea, Cindy J.; Marks, James D.

    2010-08-04

    Methods to detect botulinum toxin, the most poisonous substance known, are reviewed. Current assays are being developed with two main objectives in mind: 1) to obtain sufficiently low detection limits to replace the mouse bioassay with an in vitro assay, and 2) to develop rapid assays for screening purposes that are as sensitive as possible while requiring an hour or less to process the sample an obtain the result. This review emphasizes the diverse analytical approaches and devices that have been developed over the last decade, while also briefly reviewing representative older immunoassays to provide background and context.

  6. Laboratory diagnosis of botulinum toxin E poisoning%一起E型肉毒毒素中毒的实验室诊断

    Institute of Scientific and Technical Information of China (English)

    焦焱; 汪春翔; 葛华; 来身德

    2011-01-01

    Objective To detect the botulinum toxin in poisoned food, in order to guide the emergency treatment Method According to the method of CB/T4789.12 - 2003, it was Food Hygiene Microbiology in Examination of Clostridium Botulinum and Botuliruis Toxin, selecting the samples from poisoned food, and than tested by toxicity test Results Botulinum toxin type E existed in poisoned food. Conclusions By the test, we can clear and definite the poisoning reason quickly and provided evidences for timely accurate treatment.%目的检测中毒食品中的肉毒毒素,以指导救治。方法按中华人民共和国国家标准GB/T4789. 12-2003(食品卫生微生物学检验肉毒梭菌及肉毒毒素检验),用采集的食物中毒样品对小白鼠进行毒力检测。结果中毒食品中检出E型肉毒毒素。结论根据试验结果很快查明中毒原因,为及时救治中毒患者提供了强有力的依据。

  7. 肉毒毒素引发一起食物中毒的检测%Detection of food poisoning caused by botulinum toxin

    Institute of Scientific and Technical Information of China (English)

    秦丽云; 李云; 王晓丽; 徐保红; 柳连顺; 吕国平

    2014-01-01

    目的 对因食入自制臭豆腐后,引起家庭内食物中毒的病因进行检测.方法 按中华人民共和国国家标准GB/T 4789·12-2003《食品卫生微生物学检验肉毒梭菌及肉毒毒素检验》方法,对食物中毒的样品用小白鼠进行肉毒毒素检测.结果 测定自制臭豆腐中存在B型肉毒毒素,毒力大致为1 000 ~ 10 000 MLD/ml的范围.结论 根据试验结果查明了中毒原因,为事故的调查提供了依据.%Objective The aim of this study was to investigate the cause of a food poisoning case in a family after consuming homemade stinky tofu.Methods The botulinum toxin was examined by mice virulence test according to GB/T 47890.12-2003 microbiological examination of food hygiene (examination of Clostridium botulinum and botulinum toxin).Results The type B botulinum toxin was identified in homemade stinky tofu,and the virulence of the toxin was approximately 1000-10000 MLD/g.Conclusion The cause of this food poisoning incidence was quickly identified by the test results,and the study provided the basis for investigating food poisoning.

  8. The effect of laser and botulinum toxin in the treatment of myofascial pain and mouth opening: A randomized clinical trial.

    Science.gov (United States)

    De Carli, Bethânia Molin Giaretta; Magro, Alessandra Kuhn Dall; Souza-Silva, Bianca Núbia; Matos, Felipe de Souza; De Carli, João Paulo; Paranhos, Luiz Renato; Magro, Eduardo Dall

    2016-06-01

    This study conducted a randomized clinical trial in 15 patients, who sought care at the Dental Clinic of the University of Passo Fundo, in order to compare the use of low-level laser and botulinum toxin in the treatment of myofascial pain and whether they alter the mouth opening of patients with temporomandibular disorder. The patients were divided into two groups: the Laser group received low-level GaAlAs laser, 100mW of power at a wavelength of 830nm in continuous light emission; and the Toxin group received 30U of botulinum toxin type A (BTX-A) in the first session, and 15U after fifteen days. The assessments were performed by measuring pain with Visual Analogue Scale (VAS), and mouth opening with a digital caliper. Data were submitted to Student's t test at 5% significance level. Regarding pain symptoms, the results indicate that groups treated with laser and toxin registered 7U in VAS, at day 5 the scores were 4.75 and 4.86U, respectively. The laser worked faster (day 12) at 2.75U, and the group treated with BTX-A registered 2.86U at day 30. Both therapies investigated were effective in reducing pain, but the effect of low-level laser was faster than the use of BTX-A. Both treatments showed no statistically significant improvement in mouth opening. PMID:27045280

  9. The effect of laser and botulinum toxin in the treatment of myofascial pain and mouth opening: A randomized clinical trial.

    Science.gov (United States)

    De Carli, Bethânia Molin Giaretta; Magro, Alessandra Kuhn Dall; Souza-Silva, Bianca Núbia; Matos, Felipe de Souza; De Carli, João Paulo; Paranhos, Luiz Renato; Magro, Eduardo Dall

    2016-06-01

    This study conducted a randomized clinical trial in 15 patients, who sought care at the Dental Clinic of the University of Passo Fundo, in order to compare the use of low-level laser and botulinum toxin in the treatment of myofascial pain and whether they alter the mouth opening of patients with temporomandibular disorder. The patients were divided into two groups: the Laser group received low-level GaAlAs laser, 100mW of power at a wavelength of 830nm in continuous light emission; and the Toxin group received 30U of botulinum toxin type A (BTX-A) in the first session, and 15U after fifteen days. The assessments were performed by measuring pain with Visual Analogue Scale (VAS), and mouth opening with a digital caliper. Data were submitted to Student's t test at 5% significance level. Regarding pain symptoms, the results indicate that groups treated with laser and toxin registered 7U in VAS, at day 5 the scores were 4.75 and 4.86U, respectively. The laser worked faster (day 12) at 2.75U, and the group treated with BTX-A registered 2.86U at day 30. Both therapies investigated were effective in reducing pain, but the effect of low-level laser was faster than the use of BTX-A. Both treatments showed no statistically significant improvement in mouth opening.

  10. Botulinum Toxin Injection for Spastic Scapular Dyskinesia After Stroke: Case Series.

    Science.gov (United States)

    Hou, Saiyun; Ivanhoe, Cindy; Li, Sheng

    2015-08-01

    Spastic scapular dyskinesia after stroke is rare, which causes impaired shoulder active range of motion (ROM). To date, there has been no report about botulinum toxin injection to spastic periscapular muscles. This study presents botulinum toxin A injection for management of spastic periscapular muscles after stroke in 2 cases.This is a retrospective study of 2 cases of spastic scapular dyskinesia after stroke. Spasticity of periscapular muscles including rhomboid and lower trapezius was diagnosed by physical examination and needle electromyographic study. Botulinum toxin was injected into the spastic periscapular muscles under ultrasound imaging guidance.During the 3-week follow-up visit after injection, both patients showed increased shoulder active ROM, without any sign of scapular destabilization.The results suggest that botulinum toxin injection to spastic periscapular muscles can increase shoulder active ROM without causing scapular destabilization in patients with poststroke spastic scapular dyskinesia. PMID:26266368

  11. Efecto protector de la toxina botulínica en colgajos cutáneos The protective effect of botulinum toxin on skin flaps

    Directory of Open Access Journals (Sweden)

    C.X. Astudillo Carrera

    2011-06-01

    Full Text Available Los colgajos cutáneos de patrón vascular randomizado son de gran utilidad en Cirugía Reconstructiva; un aporte vascular adecuado es el factor más importante para la supervivencia de los mismos. Realizamos un estudio experimental levantando un total de 36 colgajos en el dorso de 9 conejos (4 en cada conejo: a 9 controles, b 9 con aplicación de toxina botulínica tipo A transoperatoria a lo largo del colgajo, c 9 con toxina botulínica tipo A aplicada 7 días antes del procedimiento quirúrgico, y, d 9 con aplicación de toxina botulínica tipo A transoperatoria más epinefrina. El análisis de los colgajos se realizó a través de imágenes digitales que se examinaron con el programa ImageJ. Efectuamos el análisis estadístico con la prueba T de Student. La evaluación final de los colgajos se realizó al séptimo día, tiempo en el cual se obtuvo: a una media de área necrótica en los controles del 72.17 ± 3.9% ; b en los que se aplicó toxina botulínica transoperatoria el porcentaje de aérea necrótica fue del 5.09 ± 1.2% p = 0.032; c en aquellos en los que la toxina botulínica se aplicó 7 días antes del procedimiento se reportó necrosis del 24.97 ± 2.7% p = 0.041 y d en los que se aplicó toxina botulínica transoperatoria más epinefrina el porcentaje de necrosis fue del 23.90 ± 3.9% p = 0.045. Concluimos que la toxina botulínica tipo A actúa como protector contra el proceso de sufrimiento-isquemia de los tejidos, debido a su acción vasodilatadora, recomendando su aplicación al inicio del procedimiento quirúrgico.Random-pattern cutaneous flaps are very useful in Reconstructive Surgery; an adequate vascular contribution is an important factor for the survival of the flaps. We designed an experimental study elevating a total of 36 dorsal flaps, divided in 4 groups: a 9 were controls, to which 0.9% physiological solution was applied, b 9 with a transoperatory application of type A botulinum toxin throughout the length of

  12. Bruxism secondary to brain injury treated with botulinum toxin-A: a case report.

    OpenAIRE

    Swinson Brian; Upile Tahwinder; Jerjes Waseem; El Maaytah Mohammed; Hopper Colin; Ayliffe Peter

    2006-01-01

    Abstract We report a successful treatment of bruxism in a patient with anoxic brain injury using botulinum toxin-A (BTX-A). On examination the mouth opening was 0 mm, no feeding was possible through the mouth. Botulinum toxin was injected into the masseter and temporalis; great improvement in trismus and bruxism was noted after 3 weeks. One further treatment improved the mouth opening on the following week and the patient was discharged from our care to be reviewed when required.

  13. Botulinum toxin treatment for lower limb extensor spasticity in chronic hemiparetic patients.

    OpenAIRE

    Hesse, S; Lücke, D; Malezic, M; Bertelt, C; Friedrich, H. (Hergen); Gregoric, M; Mauritz, K H

    1994-01-01

    Twelve chronic hemiparetic outpatients with pronounced lower limb extensor spasticity were injected with 400 units of botulinum toxin A, EMG guided into the soleus, tibialis posterior, and both heads of the gastrocnemius muscles. Botulinum toxin A caused a definite reduction of plantar flexor spasticity, in 10 patients two weeks after the injection, as assessed by the Ashworth scale. Four of the patients were able to achieve active dorsiflexion of their affected ankle. Gait analysis including...

  14. Therapeutic attenuation of mitochondrial dysfunction and oxidative stress in neurotoxin models of Parkinson's disease

    OpenAIRE

    Stack, Edward C.; Ferro, Joellyn L.; Kim, Jinho; Del Signore, Steven J.; Goodrich, Sarah; Matson, Samantha; Hunt, Bonnie B.; Cormier, Kerry; Smith, Karen; Matson, Wayne R.; Ryu, Hoon; Ferrante, Robert J.

    2008-01-01

    Therapeutic attenuation of mitochondrial dysfunction and oxidative stress in neurotoxin models of Parkinson?s disease correspondence: Corresponding author. GRECC Unit 182B, Bedford VA Medical Center, 200 Springs Road, Bedford, MA 01730, USA. Tel.: +1 781 687 2908; fax: +1 781 687 3515. (Ferrante, Robert J.) (Ferrante, Robert J.) Department of Neurology, Boston University School of Medicine - Boston--> , MA 02118--> ...

  15. Oligosaccharide composition of the neurotoxin responsive Na+ channel and the requirement of sialic acid for activity

    International Nuclear Information System (INIS)

    The neurotoxin responsive Na+ channel was purified to homogeneity in an 18% yield from a clonal cell line of mouse neuroblastoma, N-18, metabolically labeled with L-[3H]fucose. The Na+ channel, a glycoprotein, M/sub r/=200,000 (gradient 7-14% PAGE) was digested with Pronase and the glycopeptides were characterized by serial lectin affinity chromatography. greater than 90% of the oligosaccharides contained sialic acid and 18% were biantennary, 39% were triantennary and 30% tetraantennary. The glycoprotein was reconstituted into artificial phospholipid vesicles and 86Rb flux was stimulated (65%) by 200 μM veratridine and 1.2 μg of scorpion venom and was inhibited (95%) by 5 μM tetrodotoxin. The requirement of sialic acid for Na+ channel activity was demonstrated since neuraminidase (0.01 U) treatment of the reconstituted glycoprotein eliminated the response of 86Rb flux to the stimulating neurotoxins. In other experiments, treatment of N-18 cells with 10 μM swainsonine, an inhibitor of glycoprotein processing, altered the oligosaccharide composition of the Na+ channel. When the abnormally glycosylated Na+ channel was reconstituted into artificial phospholipid vesicles, 86Rb flux in response to neurotoxins was impaired. Thus, glycosylation of the polypeptide with oligosaccharides of specific composition and structure is essential for expression of the biological activity of the neurotoxin responsive Na+ channel

  16. A comparative study on three analytical methods for the determination of the neurotoxin BMAA in cyanobacteria

    NARCIS (Netherlands)

    Faassen, E.J.; Gillissen, F.; Lurling, M.

    2012-01-01

    The cyanobacterial neurotoxin ß-N-methylamino-L-alanine (BMAA) has been considered a serious health threat because of its putative role in multiple neurodegenerative diseases. First reports on BMAA concentrations in cyanobacteria were alarming: nearly all cyanobacteria were assumed to contain high B

  17. Presence of the neurotoxin BMAA in aquatic ecosystems: What do we really know?

    NARCIS (Netherlands)

    Faassen, E.J.

    2014-01-01

    The neurotoxin ß-N-methylamino-L-alanine (BMAA) is suspected to play a role in the neurological diseases amyotrophic lateral sclerosis, Alzheimer's disease, and Parkinson's disease. BMAA production by cyanobacteria has been reported and contact with cyanobacteria infested waters or consumption of aq

  18. Innovative mode of action based in vitro assays for detection of marine neurotoxins

    NARCIS (Netherlands)

    Nicolas, J.A.Y.

    2015-01-01

    Innovative mode of action based in vitro assays for detection of marine neurotoxins J. Nicolas, P.J.M. Hendriksen, T.F.H. Bovee, I.M.C.M. Rietjens Marine biotoxins are naturally occurring compounds produced by particular phytoplankton species. These toxins often accumulate in seafood and thereby rep

  19. Trans generational effects of the neurotoxin BMAA on the aquatic grazer Daphnia magna

    NARCIS (Netherlands)

    Faa