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Sample records for exfoliative toxin types

  1. Exfoliative Toxins of Staphylococcus aureus

    PubMed Central

    Bukowski, Michal; Wladyka, Benedykt; Dubin, Grzegorz

    2010-01-01

    Staphylococcus aureus is an important pathogen of humans and livestock. It causes a diverse array of diseases, ranging from relatively harmless localized skin infections to life-threatening systemic conditions. Among multiple virulence factors, staphylococci secrete several exotoxins directly associated with particular disease symptoms. These include toxic shock syndrome toxin 1 (TSST-1), enterotoxins, and exfoliative toxins (ETs). The latter are particularly interesting as the sole agents responsible for staphylococcal scalded skin syndrome (SSSS), a disease predominantly affecting infants and characterized by the loss of superficial skin layers, dehydration, and secondary infections. The molecular basis of the clinical symptoms of SSSS is well understood. ETs are serine proteases with high substrate specificity, which selectively recognize and hydrolyze desmosomal proteins in the skin. The fascinating road leading to the discovery of ETs as the agents responsible for SSSS and the characterization of the molecular mechanism of their action, including recent advances in the field, are reviewed in this article. PMID:22069631

  2. Exfoliative toxins of Staphylococcus aureus.

    PubMed

    Bukowski, Michal; Wladyka, Benedykt; Dubin, Grzegorz

    2010-05-01

    Staphylococcus aureus is an important pathogen of humans and livestock. It causes a diverse array of diseases, ranging from relatively harmless localized skin infections to life-threatening systemic conditions. Among multiple virulence factors, staphylococci secrete several exotoxins directly associated with particular disease symptoms. These include toxic shock syndrome toxin 1 (TSST-1), enterotoxins, and exfoliative toxins (ETs). The latter are particularly interesting as the sole agents responsible for staphylococcal scalded skin syndrome (SSSS), a disease predominantly affecting infants and characterized by the loss of superficial skin layers, dehydration, and secondary infections. The molecular basis of the clinical symptoms of SSSS is well understood. ETs are serine proteases with high substrate specificity, which selectively recognize and hydrolyze desmosomal proteins in the skin. The fascinating road leading to the discovery of ETs as the agents responsible for SSSS and the characterization of the molecular mechanism of their action, including recent advances in the field, are reviewed in this article.

  3. Isoelectric Focusing Patterns of Staphylococcal Exfoliative Toxin

    DTIC Science & Technology

    1983-01-01

    were. however, serologically distinct, giving reac- Electrode solutions in the gel focusing chambers sere 3 ( tions of nonidentity in Ouchterlony ...062 I 312 CURRENT MRIROBOI OG. Vol. 8 119831 TOXIN TYPE ELECTROFOCUS OUCHTERLONY ETA ETB ANODE - pH 8 0 a. ii9- 00 13 PH 6 CATHODE Fig. 2

  4. Detection of staphylococcal exfoliative toxin by slide latex agglutination.

    PubMed Central

    Murono, K; Fujita, K; Yoshioka, H

    1988-01-01

    A simple and rapid method in which slide latex agglutination was used was developed to detect the exfoliative toxin (ET) elaborated by clinical isolates. ET types A and B (ET-A and ET-B) were purified by plate gel isoelectrofocusing, and anti-ET sera were obtained by immunizing rabbits. A specific immunoglobulin G antitoxin was then prepared from the immunized rabbit sera by fast protein liquid chromatography, and latex particles were coated with the antitoxin. Of 74 staphylococcal strains isolated from patients with staphylococcal scalded skin syndrome, 61 strains were found to produce ET by the newborn mouse bioassay. All 61 strains were shown to be positive for ET-A and ET-B production by the slide latex agglutination method. The lowest concentration of ETs detected by the latex agglutination method was 0.5 microgram/ml, which was much lower than that detected by the double immunodiffusion method, with a sensitivity of 50 micrograms/ml. It is crucial to prove ET production by clinical isolates for the diagnosis and surveillance of staphylococcal scalded skin syndrome. The latex agglutination method is a sensitive, simple, and rapid test which can be used as an alternative to the newborn mouse bioassay. Images PMID:3343322

  5. Molecular and Serological Differentiation of Staphylococcal Exfoliative Toxin Synthesized Under Chromosomal and Plasmid Control

    PubMed Central

    Wiley, Bill B.; Rogolsky, M.

    1977-01-01

    Evidence for the existence of two molecular species of exfoliative toxin (ET) synthesized by phage group II Staphylococcus aureus under chromosomal and plasmid control is presented. Serological evidence that these molecular species of toxin are distinct from each other is given. The plasmid-controlled toxin was synthesized along with the chromosomally controlled toxin by the group II UT0002 strain, whereas another group II strain, UT0007, synthesized only the plasmid-controlled toxin. The molecular weight of the plasmid-controlled toxin was slightly less than that of the chromosomally controlled type and could be separated from the latter on 12.5% sodium dodecyl sulfate (SDS)-polyacrylamide slab gels. On 7.5% SDS-polyacrylamide cylindrical gels there was no hint of heterogeneity, and both ETs migrated together as a single homogeneous band. The existence of two serotypes of ET among phage group II strains complicates interpretation of previous work in this field and makes necessary the preparation of two different antigens for radioimmunobinding assays. Discovery of these ET serotypes provided an explanation for previously reported low binding by rabbit hyperimmune serum (B. Wiley, L. Glasgow, and M. Rogolsky, Infect. Immun. 13:513-520, 1976) in the radioimmunobinding test. A molecular species of ET differing from each of the other two serotypes was isolated from cultures of a phage group III S. aureus. This ET produced scalding in suckling mice and was lower in molecular weight than the ET produced under plasmid control by group II strains. Preliminary serological studies indicated that the ET in the group III strain is closely related to or possibly identical to the group II toxin produced under plasmid control. Images PMID:411758

  6. Clinical, Microbial, and Biochemical Aspects of the Exfoliative Toxins Causing Staphylococcal Scalded-Skin Syndrome

    PubMed Central

    Ladhani, Shamez; Joannou, Christopher L.; Lochrie, Denise P.; Evans, Robert W.; Poston, Susan M.

    1999-01-01

    The exfoliative (epidermolytic) toxins of Staphylococcus aureus are the causative agents of the staphylococcal scalded-skin syndrome (SSSS), a blistering skin disorder that predominantly affects children. Clinical features of SSSS vary along a spectrum, ranging from a few localized blisters to generalized exfoliation covering almost the entire body. The toxins act specifically at the zona granulosa of the epidermis to produce the characteristic exfoliation, although the mechanism by which this is achieved is still poorly understood. Despite the availability of antibiotics, SSSS carries a significant mortality rate, particularly among neonates with secondary complications of epidermal loss and among adults with underlying diseases. The aim of this article is to provide a comprehensive review of the literature spanning more than a century and to cover all aspects of the disease. The epidemiology, clinical features, potential complications, risk factors, susceptibility, diagnosis, differential diagnoses, investigations currently available, treatment options, and preventive measures are all discussed in detail. Recent crystallographic data on the toxins has provided us with a clearer and more defined approach to studying the disease. Understanding their mode of action has important implications in future treatment and prevention of SSSS and other diseases, and knowledge of their specific site of action may provide a useful tool for physiologists, dermatologists, and pharmacologists. PMID:10194458

  7. Superficial dsg2 expression limits epidermal blister formation mediated by pemphigus foliaceus antibodies and exfoliative toxins.

    PubMed

    Brennan, Donna; Hu, Ying; Medhat, Walid; Dowling, Alicia; Mahoney, My G

    2010-01-01

    Cell-cell adhesion mediated by desmosomes is crucial for maintaining proper epidermal structure and function, as evidenced by several severe and potentially fatal skin disorders involving impairment of desmosomal proteins. Pemphigus foliaceus (PF) and staphylococcal scalded skin syndrome (SSSS) are subcorneal blistering diseases resulting from loss of function of the desmosomal cadherin, desmoglein 1 (Dsg1). To further study the pathomechanism of these diseases and to assess the adhesive properties of Dsg2, we employed a recently established transgenic (Tg) mouse model expressing Dsg2 in the superficial epidermis. Neonatal Tg and wild type (WT) mice were injected with purified ETA or PF Ig. We showed that ectopic expression of Dsg2 reduced the extent of blister formation in response to both ETA and PF Ig. In response to PF Ig, we observed either a dramatic loss or a reorganization of Dsg1-alpha, Dsg1-beta, and, to a lesser extent, Dsg1-gamma, in WT mice. The Inv-Dsg2 Tg mice showed enhanced retention of Dsg1 at the cell-cell border. Collectively, our data support the role for Dsg2 in cell adhesion and suggest that ectopic superficial expression of Dsg2 can increase membrane preservation of Dsg1 and limit epidermal blister formation mediated by PF antibodies and exfoliative toxins.

  8. Superficial Dsg2 Expression Limits Epidermal Blister Formation Mediated by Pemphigus Foliaceus Antibodies and Exfoliative Toxins

    PubMed Central

    Brennan, Donna; Hu, Ying; Medhat, Walid; Dowling, Alicia; Mahoney, Mỹ G.

    2010-01-01

    Cell-cell adhesion mediated by desmosomes is crucial for maintaining proper epidermal structure and function, as evidenced by several severe and potentially fatal skin disorders involving impairment of desmosomal proteins. Pemphigus foliaceus (PF) and staphylococcal scalded skin syndrome (SSSS) are subcorneal blistering diseases resulting from loss of function of the desmosomal cadherin, desmoglein 1 (Dsg1). To further study the pathomechanism of these diseases and to assess the adhesive properties of Dsg2, we employed a recently established transgenic (Tg) mouse model expressing Dsg2 in the superficial epidermis. Neonatal Tg and wild type (WT) mice were injected with purified ETA or PF Ig. We showed that ectopic expression of Dsg2 reduced the extent of blister formation in response to both ETA and PF Ig. In response to PF Ig, we observed either a dramatic loss or a reorganization of Dsg1-α, Dsg1-β, and, to a lesser extent, Dsg1-γ, in WT mice. The Inv-Dsg2 Tg mice showed enhanced retention of Dsg1 at the cell-cell border. Collectively, our data support the role for Dsg2 in cell adhesion and suggest that ectopic superficial expression of Dsg2 can increase membrane preservation of Dsg1 and limit epidermal blister formation mediated by PF antibodies and exfoliative toxins. PMID:20631906

  9. Synthetic exfoliative toxin A and B DNA probes for detection of toxigenic Staphylococcus aureus strains.

    PubMed Central

    Rifai, S; Barbancon, V; Prevost, G; Piemont, Y

    1989-01-01

    Two methods for the detection of exfoliative toxin (ET) from Staphylococcus aureus were compared: (i) a phenotypic assay, electrosyneresis, and (ii) a genotypic assay, staphylococcal DNA hybridization with oligodeoxynucleotide probes. The probes were chosen from the previously determined sequences of serotype A and B of ET, one probe for serotype A and another for serotype B. Strains exhibiting ET production in electrosyneresis always possessed the ET gene(s). Conversely, some strains not exhibiting ET production in electrosyneresis harbored the ET gene(s). The latter strains produced levels of ET. ET-negative phage group 2 strains of S. aureus as well as tested coagulase-negative staphylococci did not possess the ET gene(s). The sensitivity of the DNA hybridization technique was 10(6) bacteria or 100 ng of genomic DNA. Images PMID:2715322

  10. Development and Evaluation of Detection Systems for Staphylococcal Exfoliative Toxin A Responsible for Scalded-Skin Syndrome

    PubMed Central

    Ladhani, Shamez; Robbie, Scott; Garratt, Richard C.; Chapple, Daniel S.; Joannou, Christopher L.; Evans, Robert W.

    2001-01-01

    Staphylococcal scalded-skin syndrome is usually diagnosed clinically by its characteristic exfoliating rash. Isolation of Staphylococcus aureus from the patient further supports the diagnosis. Several detection systems have been developed to determine whether the isolated strain produces exfoliative toxin, but none are routinely available in hospital laboratories. In a novel approach, we used computer models to predict the structure of the exfoliative toxins based on other serine proteases and to identify surface epitopes for the production of antibodies that specifically bound the exfoliative toxin A (ETA) serotype. Several rapid immunologically based diagnostic tests for ETA were developed with these antibodies and compared with existing systems. Our results showed that Western blot analysis using these antibodies was in complete correlation with PCR, which has been validated against the “gold standard” mouse model. On the other hand, the double-antibody enzyme-linked immunosorbent assay (ELISA) and Ouchterlony immunodiffusion assay gave unacceptably high false-positive results due to interference by staphylococcal protein A. This problem was successfully overcome by the development of a F(ab′)2 fragment ELISA, which was rapid and reproducible and was as sensitive and specific as PCR and Western blot analysis. The F(ab′)2 fragment ELISA is superior to existing diagnostic systems because it is quantitative, which may be related to the severity of the condition, and can detect amounts of exfoliative toxin in the picogram range directly from serum. This is the first detection system with the potential to confirm the diagnosis of staphylococcal scalded-skin syndrome from a routine blood test within 3 h of presentation. PMID:11376033

  11. Keratinocytes produce IL-6 in response to desmoglein 1 cleavage by Staphylococcus aureus exfoliative toxin A.

    PubMed

    Rolle, Cleo E; Chen, Juan; Pastar, Irena; Cardenas, Tatiana C P; Perez, Roberto; Hower, Suzanne; Ferracci, Franco; Snyder, Richard; Tomic-Canic, Marjana; Plano, Lisa R W

    2013-12-01

    Many skin infections are caused by Staphylococcus aureus, a bacterial pathogen that produces virulence factors associated with these conditions such as exfoliative toxins A and B (ETA, ETB) and the leukotoxin Panton-Valentine leukocidin (PVL). Herein, we examine the potential of skin-infecting S. aureus to produce virulence factors and their impact on the local immune response. Toxin gene profiles were generated from 188 S. aureus isolated as single infecting organisms from skin lesions and demonstrated a higher potential to express ETA, ETB, and PVL than community isolates (p < 0.001). Within the study isolate group, the prevalence of genes encoding PVL was higher among methicillin-resistant S. aureus (MRSA; n = 49), while genes encoding ETs were more prevalent in methicillin-susceptible S. aureus (MSSA; n = 139). When lesion-associated white blood cell (WBC) counts were dichotomized into high- or low-WBC-count-associated bacteria, the gene for ETA was found to be associated with a low WBC count among MSSA (p = 0.001). The ETA-induced mouse model of staphylococcal scalded skin syndrome was used to investigate the link between ETA and cytokine production. Elevated IL-6 levels in the serum and increased expression of IL-6 mRNA in the skin were detected in response to ETA exposure. These findings were recapitulated in vitro using primary human keratinocytes. Thus, S. aureus may influence the local immune response via ETA cleavage of desmoglein 1 and the induction of cutaneous IL-6 expression.

  12. Identification of a novel Staphylococcus pseudintermedius exfoliative toxin gene and its prevalence in isolates from canines with pyoderma and healthy dogs.

    PubMed

    Iyori, Keita; Hisatsune, Junzo; Kawakami, Tetsuji; Shibata, Sanae; Murayama, Nobuo; Ide, Kaori; Nagata, Masahiko; Fukata, Tsuneo; Iwasaki, Toshiroh; Oshima, Kenshiro; Hattori, Masahira; Sugai, Motoyuki; Nishifuji, Koji

    2010-11-01

    Staphylococcal exfoliative toxins are involved in some cutaneous infections in mammals by targeting desmoglein 1 (Dsg1), a desmosomal cell-cell adhesion molecule. Recently, an exfoliative toxin gene (exi) was identified in Staphylococcus pseudintermedius isolated from canine pyoderma. The aim of this study was to identify novel exfoliative toxin genes in S. pseudintermedius. Here, we describe a novel orf in the genome of S. pseudintermedius isolated from canine impetigo, whose deduced amino acid sequence was homologous to that of the SHETB exfoliative toxin from Staphylococcus hyicus (70.4%). The ORF recombinant protein caused skin exfoliation and abolished cell surface staining of Dsg1 in canine skin. Moreover, the ORF protein degraded the recombinant extracellular domains of canine Dsg1, but not Dsg3, in vitro. PCR analysis revealed that the orf was present in 23.2% (23/99) of S. pseudintermedius isolates from dogs with superficial pyoderma exhibiting various clinical phenotypes, while the occurrence in S. pseudintermedius isolates from healthy dogs was 6.1% (3/49). In summary, this newly found orf in S. pseudintermedius encodes a novel exfoliative toxin, which targets a cell-cell adhesion molecule in canine epidermis and might be involved in a broad spectrum of canine pyoderma.

  13. Emergence of Staphylococcus aureus Carrying Multiple Drug Resistance Genes on a Plasmid Encoding Exfoliative Toxin B

    PubMed Central

    Hisatsune, Junzo; Hirakawa, Hideki; Yamaguchi, Takayuki; Fudaba, Yasuyuki; Oshima, Kenshiro; Hattori, Masahira; Kato, Fuminori; Kayama, Shizuo

    2013-01-01

    We report the complete nucleotide sequence and analysis of pETBTY825, a Staphylococcus aureus TY825 plasmid encoding exfoliative toxin B (ETB). S. aureus TY825 is a clinical isolate obtained from an impetigo patient in 2002. The size of pETBTY825, 60.6 kbp, was unexpectedly larger than that of the archetype pETBTY4 (∼30 kbp). Genomic comparison of the plasmids shows that pETBTY825 has the archetype pETBTY4 as the backbone and has a single large extra DNA region of 22.4 kbp. The extra DNA region contains genes for resistance to aminoglycoside [aac(6′)/aph(2″)], macrolide (msrA), and penicillin (blaZ). A plasmid deletion experiment indicated that these three resistance elements were functionally active. We retrospectively examined the resistance profile of the clinical ETB-producing S. aureus strains isolated in 1977 to 2007 using a MIC determination with gentamicin (GM), arbekacin (ABK), and erythromycin (EM) and by PCR analyses for aac(6′)/aph(2″) and msrA using purified plasmid preparations. The ETB-producing S. aureus strains began to display high resistance to GM, which was parallel with the detection of aac(6′)/aph(2″) and mecA, after 1990. Conversely, there was no significant change in the ABK MIC during the testing period, although it had a tendency to slightly increase. After 2001, isolates resistant to EM significantly increased; however, msrA was hardly detected in ETB-producing S. aureus strains, and only five isolates were positive for both aac(6′)/aph(2″) and msrA. In this study, we report the emergence of a fusion plasmid carrying the toxin gene etb and drug resistance genes. Prevalence of the pETBTY825 carrier may further increase the clinical threat, since ETB-producing S. aureus is closely related to more severe impetigo or staphylococcal scalded-skin syndrome (SSSS), which requires a general antimicrobial treatment. PMID:24080652

  14. A novel positive regulatory element for exfoliative toxin A gene expression in Staphylococcus aureus.

    PubMed

    Sakurai, Susumu; Suzuki, Hitoshi; Hata, Toshiaki; Yoshizawa, Yukio; Nakayama, Ritsuko; Machida, Katsuhiko; Masuda, Shogo; Tsukiyama, Takashi

    2004-04-01

    A 1.4 kb positive regulatory element (ETA(exp)) that controls staphylococcal exfoliative toxin A (sETA) transcription was cloned from Staphylococcus aureus. ETA(exp) is located upstream of the cloned 5.8 kb eta gene (etaJ1) obtained from the chomosomal DNA of S. aureus ZM, the standard ETA-producing strain. The cETA prepared from an Escherichia coli transformant into which the recombinant plasmid petaJ1 (5.8 kb eta/pUC9) had been introduced was expressed at high levels in the culture supernatant and the ammonium-sulfate-precipitated culture supernatant fraction as shown by immunoblotting and the single radial immunodiffusion test. However, cETA produced by the recombinant plasmid petaJ3 containing the 1.7 kb eta sequence (etaJ3) with a 1.45 kb ETA(exp)-deficient eta fragment (1.7 kb eta/pUC9) obtained from the 5.8 kb eta sequence by subcloning was not detected in either the culture supernatant or the ammonium-sulfate-precipitated culture supernatant fraction (167-fold concentrate of the culture supernatant) by immunoblotting or the single radial immunodiffusion test. A large amount of cETA was produced by the 1.7 kb eta sequence when it was linked to ETA(exp) amplified by PCR (1.7 kb eta-ETA(exp)/pUC9), regardless of the orientation of ETA(exp) insertion. Northern blot hybridization showed lower levels of the transcripts of the 1.7 kb eta sequence than of the 5.8 kb eta sequence. The rsETA prepared from an S. aureus transformant into which the recombinant plasmid 3.4 kb eta-ETA(exp)/pYT3 (pYT3-etaJ6) had been introduced was expressed at high levels in the culture supernatant fraction as shown by the latex agglutination test. However, the agglutination titre in the culture supernatant fraction of rsETA produced by the recombinant plasmid (1.7 kb eta/pYT3) containing the 1.7 kb eta sequence carrying the 1.4 kb ETA(exp)-deficient eta fragment (pYT3-etaJ3) was 2500-4000 times lower than that of pYT3-etaJ6.

  15. Distribution of toxin genes among different spa types and phage types of animal Staphylococcus aureus.

    PubMed

    Garbacz, Katarzyna; Piechowicz, Lidia; Mroczkowska, Aneta

    2015-09-01

    We analyzed distribution of toxin genes (sea-seo, eta, etb, tst, lukS/lukF-PV) among spa types and phage types of 39 Staphylococcus aureus (S. aureus) isolates from healthy and diseased animals. All isolates turned out to be mecA negative (MSSA). Nine spa types were identified: t144 and t723 (dogs), t084 (dogs and pigs), t5447 (cat), t1491 and t008 (pigs), t002, t127 and t3478 (poultry). Seven phage types were detected, enclosed within four phage groups: I (cat), II (dogs), III (pigs) and mixed group (dogs and pigs). Three poultry spa types proved to be non-typeable by phages. Toxin genes were detected in 33 out of the 39 animal isolates. Our analysis revealed that the incidence of some toxin genes in S. aureus is host specific. Canine isolates t144 of phage group II harbored exfoliative toxin gene (eta), and porcine isolates type t1491 representing phage group III showed enterotoxin A gene (sea). The enterotoxin gene cluster (egc1) and enterotoxin gene seh were found in non-typeable isolates from chicken and in one feline isolate type t5447.

  16. Severity of Nonbullous Staphylococcus aureus Impetigo in Children Is Associated with Strains Harboring Genetic Markers for Exfoliative Toxin B, Panton-Valentine Leukocidin, and the Multidrug Resistance Plasmid pSK41

    PubMed Central

    Koning, Sander; van Belkum, Alex; Snijders, Susan; van Leeuwen, Willem; Verbrugh, Henri; Nouwen, Jan; Op ′t Veld, Mariet; van Suijlekom-Smit, Lisette W. A.; van der Wouden, Johannes C.; Verduin, Cees

    2003-01-01

    Nonbullous impetigo is a common skin infection in children and is frequently caused by Staphylococcus aureus. Staphylococcal toxins and especially exfoliative toxin A are known mediators of bullous impetigo in children. It is not known whether this is also true for nonbullous impetigo. We set out to analyze clonality among clinical isolates of S. aureus from children with nonbullous impetigo living in a restricted geographical area in The Netherlands. We investigated whether staphylococcal nasal carriage and the nature of the staphylococcal strains were associated with the severity and course of impetigo. Bacterial isolates were obtained from the noses and wounds of children suffering from impetigo. Strains were genetically characterized by pulsed-field gel electrophoresis-mediated typing and binary typing, which was also used to assess toxin gene content. In addition, a detailed clinical questionnaire was filled in by each of the participating patients. Staphylococcal nasal carriage seems to predispose the patients to the development of impetigo, and 34% of infections diagnosed in the Rotterdam area are caused by one clonal type of S. aureus. The S. aureus strains harbor the exfoliative toxin B (ETB) gene as a specific virulence factor. In particular, the numbers (P = 0.002) and sizes (P < 0.001) of the lesions were increased in patients infected with an ETB-positive strain. Additional predictors of disease severity and development could be identified. The presence of a staphylococcal plasmid encoding multiple antibiotic resistance traits, as detected by binary typing, was associated with a reduction in the cure rate. Our results recognize that a combination of staphylococcal virulence and resistance genes rather than a single gene determines the development and course of nonbullous impetigo. The identification of these microbial genetic markers, which are predictive of the severity and the course of the disease, will facilitate guided individualized antimicrobial

  17. Structural similarities and differences in Staphylococcus aureus exfoliative toxins A and B as revealed by their crystal structures.

    PubMed

    Papageorgiou, A C; Plano, L R; Collins, C M; Acharya, K R

    2000-03-01

    Staphylococcal aureus epidermolytic toxins (ETs) A and B are responsible for the induction of staphylococcal scalded skin syndrome, a disease of neonates and young children. The clinical features of this syndrome vary from localized blisters to severe exfoliation affecting most of the body surface. Comparison of the crystal structures of two subtypes of ETs-rETA (at 2.0 A resolution), rETB (at 2.8 A resolution), and an active site variant of rETA, Ser195Ala at 2.0 A resolution has demonstrated that their overall topology resembles that of a "trypsin-like" serine protease, but with significant differences at the N- and C-termini and loop regions. The details of the catalytic site in both ET structures are very similar to those in glutamate-specific serine proteases, suggesting a common catalytic mechanism. However, the "oxyanion hole," which is part of the catalytic sites of glutamate specific serine proteases, is in the closed or inactive conformation for rETA, yet in the open or active conformation for rETB. The ETs contain a unique amphipathic helix at the N-terminus, and it appears to be involved in optimizing the conformation of the catalytic site residues. Determination of the structure of the rETA catalytic site variant, Ser195Ala, showed no significant perturbation at the active site, establishing that the loss of biological and esterolytic activity can be attributed solely to disruption of the catalytic serine residue. Finally, the crystal structure of ETs, together with biochemical data and mutagenesis studies, strongly confirms the classification of these molecules as "serine proteases" rather than "superantigens."

  18. Exfoliative cytology of oral epithelial cells from patients with type 2 diabetes: cytomorphometric analysis.

    PubMed

    Rivera, César; Núñez-de-Mendoza, Camila

    2013-01-01

    This research objective is to identify cytomorphometrical changes using exfoliative cytology (EC) and later Papanicolaou (Pap) staining, for oral epithelial cells of patients with type 2 diabetes (DM2) (n = 30), while being compared to patients without the disease (n = 30). Additionally, we investigated an association between cellular changes and salivary flow levels; relationship that until now has not been reported. Results show that the cell diameter and the nuclear-cytoplasmic ratio was significantly higher compared to those patients without the disease (p ≤ 0.001 Student and Welch test). Decreased salivary flow was significantly associated with increased cell diameter and nuclear-cytoplasmic ratio (p ≤ 0.001 ANOVA with Tukey test). Evidence and clinical observations show that DM2 and decreased salivary flow are related to detectable cytomorphometrical changes in exfoliated cells, which may extend the horizon of this cytological technique.

  19. Exfoliative cytology of oral epithelial cells from patients with type 2 diabetes: cytomorphometric analysis

    PubMed Central

    Rivera, César; Núñez-de-Mendoza, Camila

    2013-01-01

    This research objective is to identify cytomorphometrical changes using exfoliative cytology (EC) and later Papanicolaou (Pap) staining, for oral epithelial cells of patients with type 2 diabetes (DM2) (n = 30), while being compared to patients without the disease (n = 30). Additionally, we investigated an association between cellular changes and salivary flow levels; relationship that until now has not been reported. Results show that the cell diameter and the nuclear-cytoplasmic ratio was significantly higher compared to those patients without the disease (p ≤ 0.001 Student and Welch test). Decreased salivary flow was significantly associated with increased cell diameter and nuclear-cytoplasmic ratio (p ≤ 0.001 ANOVA with Tukey test). Evidence and clinical observations show that DM2 and decreased salivary flow are related to detectable cytomorphometrical changes in exfoliated cells, which may extend the horizon of this cytological technique. PMID:24040475

  20. Botulinum toxin type B micromechanosensor

    PubMed Central

    Liu, W.; Montana, Vedrana; Chapman, Edwin R.; Mohideen, U.; Parpura, Vladimir

    2003-01-01

    Botulinum neurotoxin (BoNT) types A, B, E, and F are toxic to humans; early and rapid detection is essential for adequate medical treatment. Presently available tests for detection of BoNTs, although sensitive, require hours to days. We report a BoNT-B sensor whose properties allow detection of BoNT-B within minutes. The technique relies on the detection of an agarose bead detachment from the tip of a micromachined cantilever resulting from BoNT-B action on its substratum, the synaptic protein synaptobrevin 2, attached to the beads. The mechanical resonance frequency of the cantilever is monitored for the detection. To suspend the bead off the cantilever we use synaptobrevin's molecular interaction with another synaptic protein, syntaxin 1A, that was deposited onto the cantilever tip. Additionally, this bead detachment technique is general and can be used in any displacement reaction, such as in receptor-ligand pairs, where the introduction of one chemical leads to the displacement of another. The technique is of broad interest and will find uses outside toxicology. PMID:14573702

  1. Serological Screening Test for Any Botulinum Toxin Type

    DTIC Science & Technology

    1982-04-01

    1), 400 LD50 of type B toxin (2) and 80 LD50 of type E toxin (3). These botulinum toxin ELISA are based on the " double sandwich" prin- ciple but...antibody for IgG of AT but not of AT 2 1, The double sandwich approach is possible because of multiple antigenic determinants of the toxin molecule... immunodiffusion tests against homologous type antitoxin obtained by injecting a rabbit with toxoid made of a relatively crude toxin sample. All neurotoxins

  2. Detection and genetic characterization of PVL-positive ST8-MRSA-IVa and exfoliative toxin D-positive European CA-MRSA-Like ST1931 (CC80) MRSA-IVa strains in Bangladesh.

    PubMed

    Paul, Shyamal Kumar; Ghosh, Souvik; Kawaguchiya, Mitsuyo; Urushibara, Noriko; Hossain, Mohammad Akram; Ahmed, Salma; Mahmud, Chand; Jilani, Md Shariful Alam; Haq, Jalaluddin Ashraful; Ahmed, Abdullah Akhtar; Kobayashi, Nobumichi

    2014-08-01

    Severe skin lesions caused by Staphylococcus aureus infection are associated with production from bacterial cells of Panton-Valentine leukocidin (PVL), a typical virulence factor of community-acquired methicillin-resistant S. aureus (CA-MRSA), as well as other toxins represented by exfoliative toxins. Through a retrospective study of 26 S. aureus strains isolated from skin lesions of diabetic patients admitted to a hospital in Bangladesh, 2 PVL-gene-positive MRSA-IVa strains and 8 PVL-negative, exfoliative toxin D (ETD) gene (etd)-positive MRSA-IVa strains were isolated. A PVL-positive MRSA-IVa strain had a type I arginine catabolic mobile element (ACME), belonged to ST8/agr-type I/spa-type t121 (a variant of t008), and harbored blaZ, tet(K), msrA, and aph(3')-IIIa, which are mostly typical characteristics found in USA300, a predominant CA-MRSA clone in the United States. Another PVL-positive MRSA strain, belonging to ST1929 (CC88)/agr-type III/spa-type t3341, was negative for ACME, but possessed blaZ and tet(K). The etd-positive MRSA-IVa strains possessed the epidermal cell differentiation inhibitor B (EDIN-B)-encoding gene (edinB) and belonged to ST1931 (CC80)/agr-type III/spa-type t11023 (a variant of t044), which was genetic trait similar to that of the European CA-MRSA ST80 clone. However, unlike the European ST80 strains, the etd-positive MRSA strains detected in the present study harbored seb, sek, and seq, while they were negative for tet(K), aph(3')-IIIa, and fusB, showing susceptibility to fusidic acid. These findings suggested that etd-positive ST1931 MRSA strains belong to the same lineage as the European ST80 MRSA clone, evolving from a common ancestral clone via acquisition of a different pathogenicity island. This is the first report of a USA300-like MRSA-IV strain, PVL-positive ST1929 (CC88) MRSA-IV, and European ST80 CA-MRSA-like etd-positive ST1931 (CC80) MRSA-IV strains isolated in Bangladesh.

  3. Exfoliative erythroderma.

    PubMed

    Milavec-Puretić, Visnja; Zorić, Zdenka; Zidanić, Martina; Drcelić, Ada; Stajminger, Gordana

    2007-01-01

    Exfoliative erythroderma refers to the skin that is diffusely red and inflamed with varying degrees and types of scaling. There are many causes of erythroderma, but the most common are exacerbations of an underlying skin disease, drug reactions and underlying malignancies. Erythroderma is a rare, potentially serious skin condition. Protein loss in the form of desquamation and exudation is significant, resulting in hypoproteinemia. Usually more than one skin biopsy should be done. Biopsy analysis is important to rule out cutaneous T-cell lymphoma. Patients should be carefully evaluated for underlying disease. Erythroderma can represent a serious medical threat to the patient, and may require hospitalization. Various forms of exfoliative erythroderma are presented, considering the etiopathogenesis, physical findings, differential diagnosis and treatment.

  4. Evaluation of oral mucosa epithelium in type II diabetic patients by an exfoliative cytology method.

    PubMed

    Jajarm, Hassan Hosseinpour; Mohtasham, Nooshin; Moshaverinia, Maryam; Rangiani, Afsaneh

    2008-09-01

    Diabetes mellitus is a common metabolic disease that causes chronic hyperglycemia and disturbances in carbohydrate, lipid, and protein metabolism. Although diabetes can cause considerable cellular changes, this field has attracted little research. We therefore decided to evaluate the quantitative and qualitative changes in oral epithelial cells using an exfoliative cytology method. In 30 control individuals and 30 patients with type II diabetes, smears were obtained from two distinct oral sites: the buccal mucosa and tongue dorsum. The oral smears were stained using Papanicolaou solution. Quantitative and qualitative changes were evaluated in each slide. For this purpose, 50 clearly defined cells in each slide were microscopically evaluated, and photographs were subjected to computerized morphometric analysis. Cytoplasmic and nuclear areas in the diabetic group were significantly higher than in the control group. The cytoplasmic/nuclear ratio was lower in the control group. At both smear sites, the proportion of cells with nuclear changes was higher in the diabetic group. Diabetes mellitus can cause alterations in the oral epithelium that are detectable with this exfoliative cytology method. The method may be viable in evaluating this disease.

  5. Infantile botulism caused by Clostridium butyricum type E toxin.

    PubMed

    Abe, Yuichi; Negasawa, Tetsuro; Monma, Chie; Oka, Akira

    2008-01-01

    The case of a 9-month-old boy with infantile botulism caused by Clostridium butyricum type E toxin is reported. Because infantile botulism is rare in Japan, it was difficult to diagnose it at an early stage. Electrophysiologic findings were useful for the diagnosis, and were characterized by incremental responses (waxing) to short intervals and rapid repetitive nerve stimulation. A bioassay for botulism in mice indicated that the patient had botulism due to type E or F botulinum toxin. C. butyricum type E was isolated from his feces, confirming the diagnosis. This is the first known case of infantile botulism due to C. butyricum type E toxin in Japan.

  6. Botulinum toxin for meralgia paresthetica in type 2 diabetes.

    PubMed

    Dhull, Pawan; Tewari, A K; Upreti, Vimal; Prakash, M S; Hari Kumar, K V S

    2013-01-01

    Botulinum toxin has been used for a variety of neuropathic conditions in diabetes mellitus. Meralgia paresthetica is a mononeuropathy of femoral nerve seen in diabetes and obesity with an unclear etiopathogenesis. We studied the role of botulinum toxin in resistant cases of meralgia paresthetica in type 2 diabetes.

  7. Bullous impetigo in children infected with methicillin-resistant Staphylococcus aureus alone or in combination with methicillin-susceptible S. aureus: analysis of genetic characteristics, including assessment of exfoliative toxin gene carriage.

    PubMed

    Shi, Da; Higuchi, Wataru; Takano, Tomomi; Saito, Kohei; Ozaki, Kyoko; Takano, Misao; Nitahara, Yoshiyuki; Yamamoto, Tatsuo

    2011-05-01

    Among bullous impetigo isolates, exfoliative toxin (ET) gene carriage was found in 61.5% of methicillin-resistant Staphylococcus aureus (MRSA) isolates versus 90.6% of methicillin-susceptible S. aureus (MSSA) isolates. MRSA-only cases were ETB or ETA positive, while MRSA/MSSA coinfection cases were ET negative for MRSA but ETA positive for MSSA. Collagen adhesin may facilitate some MRSA infections.

  8. Bullous Impetigo in Children Infected with Methicillin-Resistant Staphylococcus aureus Alone or in Combination with Methicillin-Susceptible S. aureus: Analysis of Genetic Characteristics, Including Assessment of Exfoliative Toxin Gene Carriage▿

    PubMed Central

    Shi, Da; Higuchi, Wataru; Takano, Tomomi; Saito, Kohei; Ozaki, Kyoko; Takano, Misao; Nitahara, Yoshiyuki; Yamamoto, Tatsuo

    2011-01-01

    Among bullous impetigo isolates, exfoliative toxin (ET) gene carriage was found in 61.5% of methicillin-resistant Staphylococcus aureus (MRSA) isolates versus 90.6% of methicillin-susceptible S. aureus (MSSA) isolates. MRSA-only cases were ETB or ETA positive, while MRSA/MSSA coinfection cases were ET negative for MRSA but ETA positive for MSSA. Collagen adhesin may facilitate some MRSA infections. PMID:21430094

  9. Purification of Clostridium botulinum Type F Progenitor Toxin

    PubMed Central

    Ohishi, Iwao; Sakaguchi, Genji

    1974-01-01

    Clostridium botulinum type F progenitor toxin was purified to a homogeneous state as judged by gel filtration on Sephadex G-200, ultracentrifugation, and disc electrophoresis. The sedimentation constant, corrected to water at 20 C, of type F progenitor toxin was determined to be 10.3 and the molecular weight to be 235,000 by ultracentrifugation at pH 6.0. The purified toxin contained a toxicity of 1.2 × 108 50% lethal doses/mg of N. In agar gel double diffusion, it formed two precipitin lines at pH 6.0. The progenitor toxin of type F differs from that of type A in that it contains no hemagglutinin and from that of type E in that it is not activable. Images PMID:4615636

  10. Purification and Properties of Clostridium botulinum Type F Toxin

    PubMed Central

    Yang, K. H.; Sugiyama, H.

    1975-01-01

    Clostridium botulinum type F toxin of proteolytic Langeland strain was purified. Toxin in whole cultures was precipitated with (NH4)2SO4. Extract of the precipitate was successively chromatographed on diethylaminoethyl-cellulose at pH 6.0, O-(carboxymethyl) cellulose at pH 4.9, Sephadex G-200 at pH 8.1, quaternary aminoethyl-Sephadex at pH 4.9, and finally diethylaminoethyl-cellulose at pH 8.1. The procedure recovered 14% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, double gel diffusion serology, and isoelectric focusing. Purified toxin had a molecular weight of 150,000 by gel filtration and 155,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Specific toxicity was 9.6 × 106 mean lethal doses per absorbancy (278 nm) unit. Sub-units of 105,000 and 56,000 molecular weight are found when purified toxin is treated with a disulfide reducing agent and electrophoresed on sodium dodecyl sulfate-polyacrylamide gels. Reciprocal cross neutralizations were demonstrated when purified type F and E toxins were reacted with antitoxins which were obtained with immunizing toxoids prepared with purified toxins. Images PMID:807160

  11. Botulinum toxin type A versus botulinum toxin type B for cervical dystonia.

    PubMed

    Duarte, Gonçalo S; Castelão, Mafalda; Rodrigues, Filipe B; Marques, Raquel E; Ferreira, Joaquim; Sampaio, Cristina; Moore, Austen P; Costa, João

    2016-10-26

    This is an update of a Cochrane review first published in 2003. Cervical dystonia is the most common form of focal dystonia and is a disabling disorder characterised by painful involuntary head posturing. There are two available formulations of botulinum toxin, with botulinum toxin type A (BtA) usually considered the first line therapy for this condition. Botulinum toxin type B (BtB) is an alternative option, with no compelling theoretical reason why it might not be as- or even more effective - than BtA. To compare the efficacy, safety and tolerability of botulinum toxin type A (BtA) versus botulinum toxin type B (BtB) in people with cervical dystonia. To identify studies for this review we searched the Cochrane Movement Disorders Group Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, Embase, reference lists of articles and conference proceedings. All elements of the search, with no language restrictions, were last run in October 2016. Double-blind, parallel, randomised, placebo-controlled trials (RCTs) comparing BtA versus BtB in adults with cervical dystonia. Two independent authors assessed records, selected included studies, extracted data using a paper pro forma, and evaluated the risk of bias. We resolved disagreements by consensus or by consulting a third author. We performed meta-analyses using the random-effects model, for the comparison BtA versus BtB to estimate pooled effects and corresponding 95% confidence intervals (95% CI). No prespecified subgroup analyses were carried out. The primary efficacy outcome was improvement on any validated symptomatic rating scale, and the primary safety outcome was the proportion of participants with adverse events. We included three RCTs, all new to this update, of very low to low methodological quality, with a total of 270 participants.Two studies exclusively enrolled participants with a known positive response to BtA treatment. This raises concerns of population enrichment

  12. Treatment of a painful keloid with botulinum toxin type A.

    PubMed

    Uyesugi, Betty; Lippincott, Benjamin; Dave, Shashank

    2010-02-01

    Keloids are associated with small-fiber neuropathy and typically present with itching, pain, and allodynia. The following is a case presentation in which painful neuropathic symptoms from a keloid were treated successfully with botulinum toxin type A. To our knowledge, this is the first such case report in the literature. Further research in the use of botulinum toxin to treat keloidal pain is warranted.

  13. Anti-Idiotype Probes for Toxin Detection

    DTIC Science & Technology

    1991-09-13

    enterotoxins A and B, the exfoliative toxins A and B and toxic shock syndrome syndrome toxin-1 to activate macrophages. All the toxins tested had the...Staphylococcal enterotoxin A, toxic shock syndrome toxin-1 and both exfoliative toxins triggered C3HeB/FeJ macrophages to secrete tumor necrosis...toxins A and B and toxic shock syndrome toxin-1 to activate macrophages. All the toxins tested had the potential to stimulate tumoricidal activity in

  14. Treatment of Frey's syndrome with botulinum toxin type B.

    PubMed

    Cantarella, Giovanna; Berlusconi, Alessandra; Mele, Vincenzo; Cogiamanian, Filippo; Barbieri, Sergio

    2010-08-01

    Frey's syndrome is a frequent sequela of parotidectomy, causing facial sweating and flushing because of gustatory stimuli. Although botulinum toxin type A has become first-line therapy for Frey's syndrome, some patients become resistant. In this study, we investigated whether another serotype, botulinum toxin type B, might be an effective alternative. Case series with planned data collection. Otolaryngology department in a university hospital. Seven patients aged 30 to 68 years, with severe Frey's syndrome, underwent the Minor test and had 80 U of botulinum toxin type B per cm(2) (mean total dose, 2354 U) injected intracutaneously in the mapped area of gustatory sweating. All patients were followed up for 12 months. One month after treatment, six of the seven patients reported that gustatory sweating and flushing had resolved, and, in the remaining patient, these symptoms had decreased. The Minor test confirmed a significant improvement. The subjective benefits remained stable for six months in four patients and for nine months in the remaining three patients; 12 months after treatment, all patients still reported some improvement. Botulinum toxin type B afforded symptomatic relief in a small sample of patients with Frey's syndrome and might be considered a potential alternative to botulinum toxin type A. Copyright (c) 2010 American Academy of Otolaryngology-Head and Neck Surgery Foundation. Published by Mosby, Inc. All rights reserved.

  15. Removal of amino-terminal extracellular domains of desmoglein 1 by staphylococcal exfoliative toxin is sufficient to initiate epidermal blister formation.

    PubMed

    Nishifuji, Koji; Shimizu, Atsushi; Ishiko, Akira; Iwasaki, Toshiroh; Amagai, Masayuki

    2010-09-01

    In both bullous impetigo and staphylococcal scalded-skin syndrome (SSSS), exfoliative toxins (ETs) produced by Staphylococcus aureus cause superficial intraepidermal blisters. ETs are known to cleave specifically a single peptide bond in the extracellular domains 3 and 4 of desmoglein (Dsg) 1. However, the precise mechanisms underlying ET-induced epidermal blister formation remain poorly understood. To determine whether cleavage of Dsg1 by an ET is sufficient to induce blister formation in vivo or if the subsequent internalization of cleaved Dsg1 or other desmosomal components is required. Skin samples obtained from neonatal mice injected with ETA were analyzed by time-lapse immunofluorescence and transmission electron microscopy for desmosomal components. Epidermal blister formation was observed as early as 60 min after ETA treatment. At this time, the amino-terminal extracellular domains of Dsg1 disappeared from the surface of keratinocytes, while the cleaved carboxy-terminal domain of Dsg1 (Dsg1-C) as well as the extracellular domains of desmocollin 1 (Dsc1-N) remained on the cell surface. Half-split desmosomes with intracytoplasmic dense plaques and attached tonofilaments were recognized ultrastructurally on the split surface of keratinocytes at 60 min. Subsequent to this, Dsg1-C and Dsc1-N gradually disappeared from the surface layer of keratinocytes. Our findings suggest that the removal of amino-terminal extracellular domains of Dsg1 by ETs is sufficient to initiate epidermal blister formation in bullous impetigo and SSSS. Copyright 2010 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.

  16. In Vitro Acetylcholinesterase Inhibition by Type A Botulinum Toxin

    PubMed Central

    Marshall, R.; Quinn, L. Y.

    1967-01-01

    Type A botulinum toxin was studied for its ability to inhibit the action of acetyl-cholinesterase. The chromogenic substrate, indophenyl acetate, was used for assay of enzyme activity. Inhibition of enzyme function was detected through use of both 6.6 × 10−6 mg (20 ld50) and 6.6 × 10−10 mg (2 × 10−3ld50) of type A botulinal toxin. Control assays were performed by use of both homologous antitoxin and heterologous antitoxins (types B and E). Enzyme inhibition was effectively prevented by use of homologous antitoxin only. The inhibition noted was specific and reproducible for given substrate, enzyme, and toxin concentrations. PMID:4860916

  17. Persistence of Upper Blepharoptosis After Cosmetic Botulinum Toxin Type A.

    PubMed

    Steinsapir, Kenneth D; Groth, Michael J; Boxrud, Cynthia A

    2015-07-01

    Upper eyelid ptosis after cosmetic botulinum toxin is generally considered short-lived and responsive to apraclonidine ophthalmic drops. The authors present a series with persistent ptosis. To report a series of patients with persistent upper eyelid ptosis after cosmetic botulinum toxin. A retrospective case review series of 7 patients referred for management after developing visually significant upper eyelid ptosis after cosmetic botulinum toxin type A treatment. Patients in this series experienced persistent visually significant ptosis after cosmetic botulinum toxin lasting from 6 weeks to 13 months. Six of the 7 patients were treated with apraclonidine ophthalmic solution. Apraclonidine drops appeared to be clinically effective within 4 to 6 weeks of the resolution of ptosis. Upper eyelid ptosis after cosmetic botulinum toxin can persist for many months after treatment. Based on this series, the authors propose that apraclonidine drops can be used at the time of initial assessment to predict the relative longevity of ptosis after cosmetic botulinum toxin treatment (Level 4 evidence recommendation). After a 1-week trial, responders can be advised that ptosis is likely to resolve in 4 to 6 weeks. Nonresponders should be counseled that resolution may take longer than 6 weeks.

  18. An SOS-regulated type 2 toxin-antitoxin system.

    PubMed

    Singletary, Larissa A; Gibson, Janet L; Tanner, Elizabeth J; McKenzie, Gregory J; Lee, Peter L; Gonzalez, Caleb; Rosenberg, Susan M

    2009-12-01

    The Escherichia coli chromosome encodes seven demonstrated type 2 toxin-antitoxin (TA) systems: cassettes of two or three cotranscribed genes, one encoding a stable toxin protein that can cause cell stasis or death, another encoding a labile antitoxin protein, and sometimes a third regulatory protein. We demonstrate that the yafNO genes constitute an additional chromosomal type 2 TA system that is upregulated during the SOS DNA damage response. The yafNOP genes are part of the dinB operon, of which dinB underlies stress-induced mutagenesis mechanisms. yafN was identified as a putative antitoxin by homology to known antitoxins, implicating yafO (and/or yafP) as a putative toxin. Using phage-mediated cotransduction assays for linkage disruption, we show first that yafN is an essential gene and second that it is essential only when yafO is present. Third, yafP is not a necessary part of either the toxin or the antitoxin. Fourth, although DinB is required, the yafNOP genes are not required for stress-induced mutagenesis in the Escherichia coli Lac assay. These results imply that yafN encodes an antitoxin that protects cells against a yafO-encoded toxin and show a protein-based TA system upregulated by the SOS response.

  19. Longitudinal Phonatory Characteristics after Botulinum Toxin Type A Injection.

    ERIC Educational Resources Information Center

    Fisher, Kimberly V.; And Others

    1996-01-01

    A study investigated the long-term effects of a Botulinum Toxin Type A injection on the glottal competency of a man with adductor spasmodic dysphonia. Results suggest that change in degree of glottal adduction over time can be observed even when vocal instability is present within each recording session. (CR)

  20. Longitudinal Phonatory Characteristics after Botulinum Toxin Type A Injection.

    ERIC Educational Resources Information Center

    Fisher, Kimberly V.; And Others

    1996-01-01

    A study investigated the long-term effects of a Botulinum Toxin Type A injection on the glottal competency of a man with adductor spasmodic dysphonia. Results suggest that change in degree of glottal adduction over time can be observed even when vocal instability is present within each recording session. (CR)

  1. Clostridium perfringens type-D enterotoxaemia in cattle: the diagnostic significance of intestinal epsilon toxin.

    PubMed

    Jones, A L; Dagleish, M P; Caldow, G L

    2015-10-17

    The aims of this study were to describe 42 cases of Clostridium perfringens type-D enterotoxaemia in cattle seen between 2003 and 2014 and to determine the diagnostic value of detecting epsilon toxin in bovine intestinal content. All cases in the series had histological brain changes considered pathognomonic for C. perfringens type-D enterotoxaemia in sheep and goats and the epsilon toxin of C. perfringens was concurrently detected in the intestinal contents of 15 (36 per cent) cases. The data from the case series indicate that intestinal epsilon toxin has a sensitivity of 56 per cent compared with histology of the brain for diagnosis of bovine C. perfringens type-D enterotoxaemia. The diagnostic specificity of detecting epsilon toxin in bovine intestinal content was investigated by screening intestinal contents of 60 bovine carcases submitted for postmortem examination. Epsilon toxin was detected in 11 (18 per cent) carcases but no pathognomonic histological brain change was found in any. The specificity of intestinal epsilon toxin was estimated to be 80.4 per cent. These studies demonstrate that for a definitive diagnosis of C. perfringens type-D enterotoxaemia in cattle histological examination of the brain is essential as the presence of epsilon toxin in the intestinal contents alone is neither sensitive nor specific enough.

  2. Pharmacotherapy in Pediatric Neurogenic Bladder Intravesical Botulinum Toxin Type A

    PubMed Central

    Sager, Cristian; Burek, Carol; Durán, Victor; Corbetta, Juan Pablo; Weller, Santiago; Juan, Bortagaray; López, Juan Carlos

    2012-01-01

    When the neurogenic bladder is refractory to anticholinergics, botulinum toxin type A is used as an alternative. The neurotoxin type A reduces bladder pressure and increases its capacity and wall compliance. Additionally, it contributes to improving urinary continence and quality of life. This novel therapy is ambulatory with a low incidence of adverse effects. Due to its transitory effect, it is necessary to repeat the injections in order to sustain its therapeutic effect. In these review article we talk about Mechanism of Action, Indications, effects, administration and presentations of the Botulinum Neurotoxin Type A in pediatric patients. Also, we make references to controversial issues surrounding its use. A bibliographic search was done selecting articles and revisions from Pubmed. The key words used were botulinum toxin A, neurogenic bladder, and children. The search was limited to patients younger than 18 years of age and reports written in English in the past ten years. PMID:22720170

  3. Nuclear morphometric and morphological analysis of exfoliated buccal and tongue dorsum cells in type-1 diabetic patients.

    PubMed

    Oz, Zehra Safi; Bektas, Sibel; Battal, Fatih; Atmaca, Hulusi; Ermis, Bahri

    2014-07-01

    Diabetes mellitus type 1 that results from immunologically mediated damage to the β-cells in the pancreas. Diabetes mellitus is characterized by recurrent or persistent hyperglycemia. Hyperglycemia can be associated with salivary gland dysfunction and alterations in the oral epithelial cells. The aim of this study was to evaluate the qualitative and quantitative changes in buccal and tongue dorsum epithelial cells using an exfoliative cytology method in type 1 diabetic patients. We performed light microscopic analysis of the buccal and tongue dorsum smears in thirty type 1 diabetic patients and thirty healthy individuals. The oral smears were stained using Papanicolaou method for cytological examination and nuclear morphometric analysis. In each case, the mean nuclear area, perimeter, length, breadth, and roundness factor were evaluated in each smear using the image analysis software (Q Win, Leica™). The nuclear area, length, breadth, and perimeters were significantly higher in the diabetic group from tongue dorsum smear than that of the control group (P < 0.05). In the cytological examination, karyorrhexis-karyolysis-karyopyknosis, binucleation, nuclear membrane irregularity, cytoplasmic polymorphism, perinuclear halo were observed in oral smears with type 1 diabetic patients. Binucleation (P = 0.002) and nuclear membrane irregularity (P = 0.024) were significantly more common in buccal smears of diabetic group. Furthermore, the sensitivity of buccal mucosa was significantly higher in the diabetic group (P = 0.006). The light microscopic and nuclear morphometric study indicates that type 1 diabetes can produce morphological and nuclear morphometric changes in the oral mucosa that are noticeable with exfoliative cytology.

  4. Type C botulinum toxin causes degeneration of motoneurons in vivo.

    PubMed

    Zhao, Li-Chun; Yang, Bo; Wang, Rengang; Lipton, Stuart A; Zhang, Dongxian

    2010-01-06

    All botulinum toxins (BoNTs, types A-G) inhibit synaptic transmitter release from motoneurons, and thus result in respiratory arrest and death. Rapid treatment with anti-BoNT antibodies can prevent progression, but recovery still requires weeks on a ventilator. Even after recovery, there is a potential for persistent fatigue in some cases of botulism even years after the insult, possibly because of motoneuron dropout for previously unknown reasons. Unique among BoNTs, the C-type (BoNT/C) cleaves two proteins involved in neurotransmitter release, syntaxin and SNAP-25, and induces apoptotic cell death in cultured cerebellar neurons. It is not clear, however, whether BoNT/C also affects neurons that encounter toxin in vivo, namely motoneurons. Here, we provide experimental evidence that BoNT/C causes a slow degeneration of motoneurons both in vitro and in vivo. This novel form of BoNT/C-induced cell death may require new treatment strategies.

  5. Thermal sensitivity of Clostridium botulinum type C toxin.

    PubMed Central

    Hubálek, Z.; Halouzka, J.

    1988-01-01

    A sterile suspension containing 950 mouse LD50 per ml of type C botulinum toxin was exposed for various periods to different temperatures. The time required for the 99% (hundred-fold) reduction of toxicity was more than 5 years at -70 degrees C or -20 degrees C, 6 months at +5 degrees C, 3 weeks at +20 degrees C, 2 weeks at +28 degrees C, 2 days at +37 degrees C, 9 h at +42 degrees C, less than 30 min at +56 degrees C, less than 20 min at +60 degrees C, and below 5 min at +80 degrees C. The results suggest that Clostridium botulinum type C toxin, if produced in an ecosystem of the mild climatic zone, might persist there over the winter season and cause the intoxication of vertebrates next early spring in the absence of further microbial toxigenesis. PMID:2972554

  6. Botulinum toxin type A as treatment of partially accommodative esotropia.

    PubMed

    Flores-Reyes, E M; Castillo-López, M G; Toledo-Silva, R; Vargas-Ortega, J; Murillo-Correa, C E; Aguilar-Ruiz, A

    2016-03-01

    To determine the effectiveness of a botulinum toxin type A injection in both medial rectus muscles in patients with partially accommodative esotropia. Residual deviation and stability of strabismus were evaluated at 18 months follow up. A prospective, analytical, quasi-experimental study was conducted on a cohort of 21 patients who underwent total cycloplegic refraction and with a residual deviation of at least 14 DP. A botulinum toxin type A dose of 5 IU was injected into each medial rectus muscle for a residual deviation greater than 18 DP, with a dose of 2.5 IU being used for a deviation between 14 and 18 DP. Multivariate logistic regression analyses were performed to relate residual deviation to variables recorded as potential predictors. A total of 21 patients were included, 33.3% (n=7) males and 66.6% (n=14) females. Mean visual acuity was -.28±.25 logMAR for right eye (range 0 to -1) and -.42±.31 logMAR for left eye (range 0 to -1.3). Mean angle of residual deviation before application of botulinum toxin was 40.95±8.6DP without spectacles correction, and 22.3±7.99 DP with full cycloplegic refraction. Adverse effects were ptosis in 14.2% (n=3), diplopia 23.8% (n=5), and vertical deviation in 33% (n=7). One patient had a poor outcome, therefore required surgical treatment. At one year follow up, 85.71% of patients showed good results with esotropia of 12 DP or less, dropping to 71.43% at 18 months of follow up. Botulinum toxin type A is an effective long-term treatment with a good response in 71.43% of patients. No predictors of good response were demonstrated. Copyright © 2015 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.

  7. Cytomorphometric analysis and morphological assessment of oral exfoliated cells in type 2 diabetes mellitus and healthy individuals: A comparative study

    PubMed Central

    Sahay, Khushboo; Rehani, Shweta; Kardam, Priyanka; Kumra, Madhumani; Sharma, Rashi; Singh, Nisha

    2017-01-01

    Context: Oral exfoliative cytology is a simple, nonaggressive technique that is well accepted by patients. Therefore, it is an attractive option, which aids in the diagnosis and observation of epithelial atypias associated with oral mucosal diseases. Aims: The aim of this study was to evaluate and compare the quantitative and qualitative alterations in exfoliative smears from type 2 diabetics and healthy individuals. Patients and Methods: The study includes 30 type 2 diabetics and 30 healthy persons of both sexes. PAP and hematoxylin and eosin (H and E) stained smears were prepared from buccal mucosa (BM), tongue (T), floor of the mouth (FOM), and palate (P). Under a light microscope, 50 clearly defined unfolded epithelial cells were quantitatively evaluated for cellular area (CA), nuclear area (NA), and cellular-to-nuclear area ratio (CA:NA) and assessed for morphological features. Statistical Analysis: Collected data was manually entered into the Statistical Package for the Social Sciences version 13.5 for analysis. Student's t-test was used at 95% confidence interval. Results: Quantitative assessment of the overall mean CA was less, mean NA was more, and mean CA:NA was less in diabetics than that in healthy persons at all the four sites. Diabetic oral cells showed qualiative cytoplasmic and nuclear alterations: cytoplasmic vacuoles, karyorrhexis, karyolysis, pyknosis, peri-nuclear halo, binucleation, nuclear vacuoles, inflammation, and microbial colonies. Conclusion: Oral cytology from type 2 diabetics is associated with detectable cytomorphological changes with alteration in size of the cell and nucleus, which is site specific, indicating epithelial cell degeneration in cytoplasm and nucleus. PMID:28182082

  8. Characterization of six type A strains of Clostridium botulinum that contain type B toxin gene sequences.

    PubMed

    Kirma, Nameer; Ferreira, Joseph L; Baumstark, Barbara R

    2004-02-16

    Six Clostridium botulinum isolates exhibiting type A toxicity as measured by the mouse bioassay were found to contain both type A and type B neurotoxin DNA sequences. The six strains were divided into three groups based on the DNA sequence of the type B neurotoxin gene. Members of each group exhibited 100% sequence identity over the 3876 bp type B toxin open reading frame. The type B toxin sequence of all groups differed at more than 60 positions when compared to the BGB control strain.

  9. Exfoliative dermatitis.

    PubMed

    Karakayli, G; Beckham, G; Orengo, I; Rosen, T

    1999-02-01

    Exfoliative dermatitis, also known as erythroderma, is an uncommon but serious skin disorder that family physicians must be able to recognize and treat appropriately. Although the etiology is often unknown, exfoliative dermatitis may be the result of a drug reaction or an underlying malignancy. The approach to treatment should include discontinuation of any potentially causative medications and a search for any underlying malignancy. One of the most common malignancies associated with exfoliative dermatitis is cutaneous T-cell lymphoma, which may not manifest for months or even years after the onset of the skin condition. Hospitalization is usually necessary for initial evaluation and treatment. In the hospital, special attention must be given to maintaining temperature control, replacing lost fluids and electrolytes, and preventing and treating infection. The long-term prognosis is good in patients with drug-induced disease, although the course tends to be remitting and relapsing in idiopathic cases. The prognosis of cases associated with malignancy typically depends on the outcome of the underlying malignancy.

  10. Clostridium perfringens type A-E toxin plasmids.

    PubMed

    Freedman, John C; Theoret, James R; Wisniewski, Jessica A; Uzal, Francisco A; Rood, Julian I; McClane, Bruce A

    2015-05-01

    Clostridium perfringens relies upon plasmid-encoded toxin genes to cause intestinal infections. These toxin genes are associated with insertion sequences that may facilitate their mobilization and transfer, giving rise to new toxin plasmids with common backbones. Most toxin plasmids carry a transfer of clostridial plasmids locus mediating conjugation, which likely explains the presence of similar toxin plasmids in otherwise unrelated C. perfringens strains. The association of many toxin genes with insertion sequences and conjugative plasmids provides virulence flexibility when causing intestinal infections. However, incompatibility issues apparently limit the number of toxin plasmids maintained by a single cell.

  11. Clostridium perfringens type A–E toxin plasmids

    PubMed Central

    Freedman, John C.; Theoret, James R.; Wisniewski, Jessica A.; Uzal, Francisco A.; Rood, Julian I.; McClane, Bruce A.

    2014-01-01

    Clostridium perfringens relies upon plasmid-encoded toxin genes to cause intestinal infections. These toxin genes are associated with insertion sequences that may facilitate their mobilization and transfer, giving rise to new toxin plasmids with common backbones. Most toxin plasmids carry a transfer of clostridial plasmids locus mediating conjugation, which likely explains the presence of similar toxin plasmids in otherwise unrelated C. perfringens strains. The association of many toxin genes with insertion sequences and conjugative plasmids provides virulence flexibility when causing intestinal infections. However, incompatibility issues apparently limit the number of toxin plasmids maintained by a single cell. PMID:25283728

  12. Clinical resistance to three types of botulinum toxin type A in aesthetic medicine.

    PubMed

    Stephan, Farid; Habre, Maya; Tomb, Roland

    2014-12-01

    Botulinum toxin injections have become the most frequent noninvasive cosmetic procedure carried out worldwide. Botulinum toxin has also multiple other indications in different medical fields. However, with the repetition of injections, a new concern has emerged: clinical resistance and loss of effectiveness of the treatment. After reporting a case of primary nonresponsiveness to three types of botulinum toxin type A injections, we conducted a review about all factors leading to the primary or secondary nonresponsiveness, as well as the factors affecting the immunogenicity of this neurotoxin. Most of the reports and studies focused on secondary resistance to botulinum toxin (BT) and the neurotoxin immunogenicity; primary nonresponsiveness was rarely reported. Factors leading to primary or secondary resistance to BT injections were numerous. In the majority of the studies, development of neutralizing antibodies to botulinum toxin was considered responsible of the induced clinical resistance. Patients should be aware of this rising concern as well as clinicians who should learn how to minimize the risk of resistance development, sparing the patients more invasive treatment modalities. Further studies related to botulinum toxin resistance are needed.

  13. Evidence for antinociceptive activity of botulinum toxin type A in pain management.

    PubMed

    Aoki, K Roger

    2003-01-01

    The neurotoxin, botulinum toxin type A, has been used successfully, in some patients, as an analgesic for myofascial pain syndromes, migraine, and other headache types. The toxin inhibits the release of the neurotransmitter, acetylcholine, at the neuromuscular junction thereby inhibiting striated muscle contractions. In the majority of pain syndromes where botulinum toxin type A is effective, inhibiting muscle spasms is an important component of its activity. Even so, the reduction of pain often occurs before the decrease in muscle contractions suggesting that botulinum toxin type A has a more complex mechanism of action than initially hypothesized. Current data points to an antinociceptive effect of botulinum toxin type A that is separate from its neuromuscular activity. The common biochemical mechanism, however, remains the same between botulinum toxin type A's effect on the motor nerve or the sensory nerve: enzymatic blockade of neurotransmitter release. The antinociceptive effect of the toxin was reported to block substance P release using in vitro culture systems. The current investigation evaluated the in vivo mechanism of action for the antinociceptive action of botulinum toxin type A. In these studies, botulinum toxin type A was found to block the release of glutamate. Furthermore, Fos, a product of the immediate early gene, c-fos, expressed with neuronal stimuli was prevented upon peripheral exposure to the toxin. These findings suggest that botulinum toxin type A blocks peripheral sensitization and, indirectly, reduces central sensitization. The recent hypothesis that migraine involves both peripheral and central sensitization may help explain how botulinum toxin type A inhibits migraine pain by acting on these two pathways. Further research is needed to determine whether the antinociceptive mechanism mediated by botulinum toxin type A affects the neuronal signaling pathways that are activated during migraine.

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

    PubMed Central

    Brashear, Allison

    2009-01-01

    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 the studies of botulinum toxin type A to treat cervical dystonia. PMID:19707390

  15. Different types of toxins targeting TRPV1 in pain.

    PubMed

    Min, Jia-Wei; Liu, Wan-Hong; He, Xiao-Hua; Peng, Bi-Wen

    2013-09-01

    The transient receptor potential vanilloid 1(TRPV1) channels are members of the transient receptor potential (TRP) superfamily. Members of this family are expressed in primary sensory neurons and are best known for their role in nociception and sensory transmission. Multiple painful stimuli can activate these channels. In this review, we discussed the mechanisms of different types of venoms that target TRPV1, such as scorpion venom, botulinum neurotoxin, spider toxin, ciguatera fish poisoning (CFP) and neurotoxic shellfish poisoning (NSP). Some of these toxins activate TRPV1; however, some do not. Regardless of TRPV1 inhibition or activation, they occur through different pathways. For example, BoNT/A decreases TRPV1 expression levels by blocking TRPV1 trafficking to the plasma membrane, although the exact mechanism is still under debate. Vanillotoxins from tarantula (Psalmopoeus cambridgei) are proposed to activate TRPV1 via interaction with a region of TRPV1 that is homologous to voltage-dependent ion channels. Here, we offer a description of the present state of knowledge for this complex subject.

  16. Relationship of bacteriophages to alpha toxin production in Clostridium novyi types A and B.

    PubMed Central

    Eklund, M W; Poysky, F T; Peterson, M E; Meyers, J A

    1976-01-01

    The relationship of specific bacteriophages to the production of the lethal alpha toxin in Clostridium novyi types A and B was investigated. When type A strain 5771 reverted to the phage-sensitive state, it ceased to produce alpha toxin but continued to produce the gamma and epsilon antigens. This "nontoxigenic" culture, therefore, more closely resembled C. botulinum types C and D than the other C. novyi types. Phage-sensitive type B strains also ceased to produce the alpha toxin but continued to produce the beta toxin, and therefore very colesly resembled C. novyi type D (C. haemolyticum). Alpha toxin was again produced when the phage-sensitive cultures were reinfected with the respective tox+ phages. Alpha toxin production could also be induced in the "nontoxigenic" phage-sensitive derivatives from type B strain 8024 by tox+ phages isolated from other strains of type B. tox- phages were also isolated, but they did not affect alpha toxin production. The tox+ phages also caused a marked change in the colonial morphology of type B strains. In this report we present evidence that alpha toxin production by C. novyi type A strain 5771 and type B strain 8024 depends upon the continued presence and participation of specific bacteriophages designated as NA1tox+ and NB1tox+, respectively. Images PMID:965096

  17. Protection by atropine against synergistic lethal effects of the Angusticeps-type toxin F7 from eastern green mamba venom and toxin I from black mamba venom.

    PubMed

    Lee, C Y; Chen, Y M; Joubert, F J

    1982-01-01

    A large dose of atropine protected mice effectively from death due to synergistic effects of an Angusticeps-type toxin, F7, isolated from eastern green mamba venom and a trypsin inhibitor homologue, toxin I, isolated from black mamba venom. Neostigmine potentiated the combined toxicity of these two toxins. It is concluded that the synergistic lethal effect of these two mamba toxins is due to massive release of acetylcholine in vivo.

  18. Distinct type I and type II toxin-antitoxin modules control Salmonella lifestyle inside eukaryotic cells.

    PubMed

    Lobato-Márquez, Damián; Moreno-Córdoba, Inmaculada; Figueroa, Virginia; Díaz-Orejas, Ramón; García-del Portillo, Francisco

    2015-03-20

    Toxin-antitoxin (TA) modules contribute to the generation of non-growing cells in response to stress. These modules abound in bacterial pathogens although the bases for this profusion remain largely unknown. Using the intracellular bacterial pathogen Salmonella enterica serovar Typhimurium as a model, here we show that a selected group of TA modules impact bacterial fitness inside eukaryotic cells. We characterized in this pathogen twenty-seven TA modules, including type I and type II TA modules encoding antisense RNA and proteinaceous antitoxins, respectively. Proteomic and gene expression analyses revealed that the pathogen produces numerous toxins of TA modules inside eukaryotic cells. Among these, the toxins HokST, LdrAST, and TisBST, encoded by type I TA modules and T4ST and VapC2ST, encoded by type II TA modules, promote bacterial survival inside fibroblasts. In contrast, only VapC2ST shows that positive effect in bacterial fitness when the pathogen infects epithelial cells. These results illustrate how S. Typhimurium uses distinct type I and type II TA modules to regulate its intracellular lifestyle in varied host cell types. This function specialization might explain why the number of TA modules increased in intracellular bacterial pathogens.

  19. Phospholipid scramblase 1 mediates type i interferon-induced protection against staphylococcal α-toxin.

    PubMed

    Lizak, Miroslaw; Yarovinsky, Timur O

    2012-01-19

    The opportunistic gram-positive pathogen Staphylococcus aureus is a leading cause of pneumonia and sepsis. Staphylococcal α-toxin, a prototypical pore-forming toxin, is a major virulence factor of S. aureus clinical isolates, and lung epithelial cells are highly sensitive to α-toxin's cytolytic activity. Type I interferon (IFN) signaling activated in response to S. aureus increases pulmonary cell resistance to α-toxin, but the underlying mechanisms are uncharacterized. We show that IFNα protects human lung epithelial cells from α-toxin-induced intracellular ATP depletion and cell death by reducing extracellular ATP leakage. This effect depends on protein palmitoylation and induction of phospholipid scramblase 1 (PLSCR1). IFNα-induced PLSCR1 associates with the cytoskeleton after exposure to α-toxin, and cellular depletion of PLSCR1 negates IFN-induced protection from α-toxin. PLSCR1-deficient mice display enhanced sensitivity to inhaled α-toxin and an α-toxin-producing S. aureus strain. These results uncover PLSCR1 activity as part of an innate protective mechanism to a bacterial pore-forming toxin. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Phospholipid scramblase 1 mediates type I interferon-induced protection against staphylococcal α-toxin

    PubMed Central

    Lizak, Miroslaw; Yarovinsky, Timur O.

    2012-01-01

    Summary The opportunistic gram-positive pathogen Staphylococcus aureus is a leading cause of pneumonia and sepsis. Staphylococcal α-toxin, a prototypical pore-forming toxin, is a major virulence factor of S. aureus clinical isolates and lung epithelial cells are highly sensitive to α-toxin's cytolytic activity. Type I interferon (IFN) signaling activated in response to S. aureus increases pulmonary cell resistance to α-toxin but the underlying mechanisms are uncharacterized. We show that IFNα protects human lung epithelial cells from α-toxin-induced intracellular ATP depletion and cell death by reducing extracellular ATP leakage. This effect depends on protein palmitoylation and induction of phospholipid scramblase 1 (PLSCR1). IFNα-induced PLSCR1 associates with the cytoskeleton after exposure to α-toxin and cellular depletion of PLSCR1 negates IFN-induced protection from α-toxin. PLSCR1-deficient mice display enhanced sensitivity to inhaled α-toxin and an α-toxin-producing S. aureus strain. These results uncover PLSCR1 activity as part of an innate protective mechanism to a bacterial pore-forming toxin. PMID:22264514

  1. Rapid Detection and Quantitative Estimation of Type A Botulinum Toxin by Electroimmunodiffusion

    PubMed Central

    Miller, Carol A.; Anderson, Arthur W.

    1971-01-01

    An experimental system is described for the detection and quantitative estimation of type A botulinum toxin by electroimmunodiffusion. The method is shown to be rapid, specific, and quantitative. As little as 14 mouse LD50 per 0.1 ml of type A toxin was detected within 2 hr. When applied to experimentally contaminated foods such as canned tuna, pumpkin, spinach, green beans, and sausage, the technique detected botulinum toxin rapidly and identified it as to type and quantity. A specific rabbit type A antitoxin was produced for this in vitro system since the equine antitoxin (Center for Disease Control) tested in this experiment was found to be unsuitable. Images PMID:5005291

  2. Experience with botulinum toxin type A in medically intractable pediatric chronic daily headache.

    PubMed

    Ahmed, Karman; Oas, Kimberly Hall; Mack, Kenneth J; Garza, Ivan

    2010-11-01

    In adults, botulinum toxin type A has been studied as a potentially effective treatment for chronic daily headache. For pediatric chronic daily headache, the literature evaluating efficacy of botulinum toxin type A is sparse, with no studies assessing tolerability. The purpose of this retrospective case series study was to assess tolerability and efficacy of botulinum toxin type A in the treatment of pediatric chronic daily headache. The series comprises 10 patients (ages 11-17 years) who received a standard 100-unit dose of onabotulinumtoxinA (trade name, Botox) for refractory chronic daily headache. Attention was given to therapeutic history, efficacy, and tolerability. The patients had attempted an average of 8.0 ± 2.40 S.D. therapies prior to botulinum toxin type A. Most patients reported adverse events from at least one of these prior medications. With botulinum toxin type A, four patients (40%) reported subjective but clinically meaningful relief, consisting of a decrease in headache intensity, and two patients additionally noted a decrease in headache frequency. The four responders noted improvements in quality of life. Three patients experienced minor adverse events from botulinum toxin type A. This case series suggests that botulinum toxin type A can be well tolerated and may be a useful therapeutic in pediatric patients with highly medically intractable chronic daily headache. Copyright © 2010 Elsevier Inc. All rights reserved.

  3. Type VI Secretion System Toxins Horizontally Shared between Marine Bacteria

    PubMed Central

    Salomon, Dor; Klimko, John A.; Trudgian, David C.; Kinch, Lisa N.; Grishin, Nick V.; Mirzaei, Hamid; Orth, Kim

    2015-01-01

    The type VI secretion system (T6SS) is a widespread protein secretion apparatus used by Gram-negative bacteria to deliver toxic effector proteins into adjacent bacterial or host cells. Here, we uncovered a role in interbacterial competition for the two T6SSs encoded by the marine pathogen Vibrio alginolyticus. Using comparative proteomics and genetics, we identified their effector repertoires. In addition to the previously described effector V12G01_02265, we identified three new effectors secreted by T6SS1, indicating that the T6SS1 secretes at least four antibacterial effectors, of which three are members of the MIX-effector class. We also showed that the T6SS2 secretes at least three antibacterial effectors. Our findings revealed that many MIX-effectors belonging to clan V are “orphan” effectors that neighbor mobile elements and are shared between marine bacteria via horizontal gene transfer. We demonstrated that a MIX V-effector from V. alginolyticus is a functional T6SS effector when ectopically expressed in another Vibrio species. We propose that mobile MIX V-effectors serve as an environmental reservoir of T6SS effectors that are shared and used to diversify antibacterial toxin repertoires in marine bacteria, resulting in enhanced competitive fitness. PMID:26305100

  4. Frontal Alopecia after Repeated Botulinum Toxin Type A Injections for Forehead Wrinkles: An Underestimated Entity?

    PubMed Central

    Di Pietro, Antonino; Piraccini, Bianca Maria

    2016-01-01

    Background Injections of botulinum toxin type A in the forehead have never been reported to cause hair side effects. Objective The aim of this paper is to report a new type of alopecia, which we have seen in women undergoing periodic injections of botulinum toxin type A for forehead wrinkles, and to differentiate it from other types of hair loss. Methods We conducted an observational study on 5 females recruited from a private and an institutional practice who complained of progressive recession of the hairline after periodic injections of botulinum toxin type A in the forehead. Results Alopecia of the frontal hairline was evident in all 5 patients, with absence of skin atrophy or scarring and progressive hair miniaturization at trichoscopy. Conclusion Dermatologists should be aware of the possible occurrence of frontal alopecia after repeated injections of botulinum toxin type A for forehead wrinkles. PMID:27843928

  5. The effects of botulinum toxin type A on muscle blood perfusion and metabolism.

    PubMed

    Matic, Damir B; Lee, Ting Y; Wells, R Glenn; Gan, Bing S

    2007-12-01

    Botulinum toxin type A is approved by the U.S. Food and Drug Administration for the treatment of facial rhytides. However, the complete spectrum of action of botulinum toxin A has not yet been completely defined. Little is known about the metabolism of muscle after botulinum toxin A injection. This information may give insight into the additional effects botulinum toxin A may have on muscle. The authors assessed the influence of botulinum toxin A on the metabolism of muscle using dynamic investigative techniques. Twenty New Zealand White rabbits were divided into control, paralysis, and sham groups. Masseter muscle paralysis was achieved with botulinum toxin A. Dynamic computed tomographic and positron emission tomographic scans were obtained. Masseter muscle blood flow, blood volume, permeability surface, and mean transit time and glucose uptake were measured. Eighteen animals completed the study. Masseter blood perfusion showed consistent results across all parameters. Blood flow, blood volume, and permeability surface were significantly increased at weeks 4 and 8 on the paralyzed side. Mean transit time at week 4 was decreased on the paralyzed side. Positron emission tomographic scans showed that injected muscles in the botulinum toxin A group tended to have increased glucose uptake compared with untreated muscles. Botulinum toxin A injection increases muscle blood perfusion parameters and glucose uptake for a transient period. This increase is similar in duration to the known interval of botulinum toxin A-induced paralysis. These changes have been identified in a dynamic fashion and may represent changes in calcitonin gene-related peptide release.

  6. The Crystal Structure of Shiga Toxin Type 2 with Bound Disaccharide Guides the Design of a Heterobifunctional Toxin Inhibitor*

    PubMed Central

    Jacobson, Jared M.; Yin, Jiang; Kitov, Pavel I.; Mulvey, George; Griener, Tom P.; James, Michael N. G.; Armstrong, Glen; Bundle, David R.

    2014-01-01

    Shiga toxin type 2 (Stx2a) is clinically most closely associated with enterohemorrhagic E. coli O157:H7-mediated hemorrhagic colitis that sometimes progresses to hemolytic-uremic syndrome. The ability to express the toxin has been acquired by other Escherichia coli strains, and outbreaks of food poisoning have caused significant mortality rates as, for example, in the 2011 outbreak in northern Germany. Stx2a, an AB5 toxin, gains entry into human cells via the glycosphingolipid receptor Gb3. We have determined the first crystal structure of a disaccharide analog of Gb3 bound to the B5 pentamer of Stx2a holotoxin. In this Gb3 analog, α-GalNAc replaces the terminal α-Gal residue. This co-crystal structure confirms previous inferences that two of the primary binding sites identified in the B5 pentamer of Stx1 are also functional in Stx2a. This knowledge provides a rationale for the synthesis and evaluation of heterobifunctional antagonists for E. coli toxins that target Stx2a. Incorporation of GalNAc Gb3 trisaccharide in a heterobifunctional ligand with an attached pyruvate acetal, a ligand for human amyloid P component, and conjugation to poly[acrylamide-co-(3-azidopropylmethacrylamide)] produced a polymer that neutralized Stx2a in a mouse model of Shigatoxemia. PMID:24225957

  7. Genotoxicity evaluation of metformin and glimepiride by micronucleus assay in exfoliated urothelial cells of type 2 diabetes mellitus patients.

    PubMed

    Harishankar, M K; Logeshwaran, S; Sujeevan, S; Aruljothi, K N; Dannie, M A; Devi, A

    2015-09-01

    Micronucleus (MN) assay was performed on the exfoliated urothelial cells to detect the genotoxic effects of the anti-hyperglycemic drugs, metformin and glimepiride in T2DM patients and to use it as a biomarker for DNA damage by assessing the frequency of micronuclei in the exfoliated urothelial cells. A total of 201 subjects (147 T2DM patients & 54 Normal cases) were selected from diverse age groups (25-75 years) and the mean MN frequency was examined per 1000 cells in all the subjects. Relative to the control group (5.02 ± 1.01), an increased MN frequency was observed in females (26.15 ± 2.15) when compared to males (23.08 ± 2.09) in T2DM patients. Further analysis showed that there was a profound increase in the number of MN in the patients using metformin alone (23.02 ± 4.44), or combination of metformin & glimepiride (24.98 ± 2.87) than to the subjects using glimepiride alone (17.52 ± 3.28). It has been proven by this simple, reliable and non-invasive method that metformin has a potential role in causing genotoxicity and that the MN observed in exfoliated urothelial cells could be used as a reliable biomarker in monitoring the genotoxic risk of the anti-hyperglycemic drugs.

  8. Effects of botulinum toxin type D on secretion of tumor necrosis factor from human monocytes

    SciTech Connect

    Imamura, K.; Spriggs, D.; Ohno, T.; Kufe, D.

    1989-05-01

    Botulinum toxins are potent neurotoxins which block the release of neurotransmitters. The effects of these toxins on hematopoietic cells, however, are unknown. Monocytes secrete a variety of polypeptide growth factors, including tumor necrosis factor (TNF). In the study reported here, the effects of botulinum toxin type D on the secretion of TNF from human monocytes were examined. The results demonstrate that biotulinum toxin type D inhibits the release of TNF from monocytes activated by lipopolysaccharide (LPS) but not by 12-O-tetradecanoylphorbol-13-acetate. Botulinum toxin type D had no detectable effect on intracellular TNF levels in LPS-treated monocytes, indicating that the effects of this toxin involve the secretory process. This inhibitory effect of botulinum toxin type D on TNF secretion from LPS-treated monocytes was partially reversed by treatment with 12-O-tetradecanoylphorbol-13-acetate or introduction of guanosine 5'-(/gamma/-thio)t-riphosphate into these cells. The results demonstrate that TNF secretion is regulated by at least two distinct guanine nucleotide-binding proteins, one responsible for the activation of phospholiphase C and another which acts as a substrate for botulinum toxin type D. ADP-ribosylation of monocyte membranes by botulinum toxin type D demonstrated the presence of three substrates with M/sub r/s of 45,000, 21,000, and 17,000. While the role of these substrates in exocytosis is unknown, the results suggest that the M/sub r/ 21,000 substrate is involved in a process other than TNF secretion.

  9. PURIFICATION AND MOLECULAR WEIGHT DETERMINATION OF CLOSTRIDIUM BOTULINUM TYPE E TOXIN

    PubMed Central

    Gerwing, Julia; Dolman, Claude E.; Reichmann, M. E.; Bains, Hardial S.

    1964-01-01

    Gerwing, Julia (The University of British Columbia, Vancouver, British Columbia, Canada), Claude E. Dolman, M. E. Reichmann, and Hardial S. Bains. Purification and molecular weight determination of Clostridium botulinum type E toxin. J. Bacteriol. 88:216–219. 1964.—A method was developed whereby type E botulinus toxin can be obtained in a highly purified state by elution through acidified diethylaminoethyl-cellulose columns. The material thus isolated appears to be electrophoretically and ultracentrifugally homogeneous. A molecular weight of 18,600 was calculated for the toxin. Images PMID:14197891

  10. Botulinum Toxin Type A (BOTOX) for Refractory Myofascial Pelvic Pain

    PubMed Central

    ADELOWO, Amos; HACKER, Michele R.; SHAPIRO, Alex; Modest, Anna Merport; ELKADRY, Eman

    2013-01-01

    Objective To assess intralevator Botulinum toxin type A (Botox) injections for refractory myofascial pelvic pain with short tight pelvic floor. Methods Retrospective cohort study of all women with intralevator Botox injection (100-300 Units) from 2005 through 2010 for refractory myofascial pelvic pain. Primary outcomes were self-reported pain on palpation and symptom improvement. Secondary outcomes included post-injection complications and repeat injection. Pain was assessed during digital palpation of the pelvic floor muscles using a scale of 0-10, with 10 being the worst possible pain. Follow-up occurred at <6 weeks post-injection and again at ≥ 6 weeks. Data are presented as median (interquartile range) or proportion. Results Thirty-one patients met eligibility criteria; 2 were lost to follow up and excluded. Median age was 55.0 years (38.0-62.0). Before Botox injection, median pain score was 9.5 (8.0-10.0). Twenty-nine patients (93.5%) returned for the first follow-up visit; 79.3% reported improvement in pain, while 20.7% reported no improvement. Median pain with levator palpation was significantly lower than before injection (P<0.0001). Eighteen women (58.0%) had a second follow-up visit with a median pain score that remained lower than before injection (P<0.0001). Fifteen (51.7%) women elected to have repeat Botox injection; the median time to repeat injection was 4.0 (3.0-7.0) months. Three (10.3%) women developed de-novo urinary retention, 2 (6.9%) reported fecal incontinence and 3 (10.3%) reported constipation and/or rectal pain; all side effects resolved spontaneously. Conclusions Intralevator injection of Botox demonstrates effectiveness in women with refractory myofascial pelvic pain with few, self-limiting adverse effects. PMID:23982578

  11. Botulinum toxin type a in the treatment of children with congenital muscular torticollis.

    PubMed

    Oleszek, Joyce L; Chang, Nicki; Apkon, Susan D; Wilson, Pamela E

    2005-10-01

    This is a retrospective case series describing the use of botulinum toxin type A in the treatment of children with congenital muscular torticollis who fail to progress with conservative management. A total of 27 children with congenital muscular torticollis, 6-18 mos of age, received 30 botulinum toxin type A injections into their sternocleidomastoid or upper trapezius muscle, or both, at a pediatric tertiary care center between 1995 and 2001. Three children received repeat injections. Twenty of 27 children (74%) had improved cervical rotation or head tilt after the injections, and 2 of 27 (7%) experienced transient adverse events, specifically, mild dysphagia and neck weakness. This series suggests that botulinum toxin type A may be a safe and effective treatment option for children with congenital muscular torticollis who are unresponsive to a traditional regimen of physical therapy and a home program. A prospective, randomized controlled trial is necessary to definitively assess the role of botulinum toxin type A in this population.

  12. Ranula successfully treated by botulinum toxin type A: report of 3 cases.

    PubMed

    Chow, Tam-Lin; Chan, Sharon W W; Lam, Siu-Ho

    2008-01-01

    The conventional treatment of ranula is surgical procedure. We report an innovative method for ranula by using botulinum toxin type A on 3 patients. All 3 cases of ranula resolved after this minimally invasive therapy. The treatment complication was minimal.

  13. Beneficial effects of botulinum toxin type a for patients with painful tic convulsif.

    PubMed

    Micheli, Federico; Scorticati, María Clara; Raina, Gabriela

    2002-01-01

    Botulinum toxin is a well-known therapy for patients with diverse movement disorders. Its application has been extended to other disorders. Here, we document the case of a 70-year-old man with hemifacial spasm associated to trigeminal neuralgia secondary to an ectatic basilar artery. He was treated with botulinum toxin type A, 2.5 mouse units over five sites at the orbicularis oculi and one over the buccinator muscle. After botulinum toxin injections, relief was gained not only from twitching but also from pain. When the effects of the toxin vanished, spasms and pain recurred. Further infiltrations were given every 12 weeks following the same response pattern. This observation further validates the increasing role of botulinum toxin in pain management.

  14. Purification and characterization of alpha-toxin produced by Clostridium novyi type A.

    PubMed Central

    Ball, D W; Van Tassell, R L; Roberts, M D; Hahn, P E; Lyerly, D M; Wilkins, T D

    1993-01-01

    Our study describes the production, purification, and properties of alpha-toxin from Clostridium novyi type A 19402. The bacterium produced maximal amounts of alpha-toxin when grown at 37 degrees C for 72 h in dialysis flask cultures containing brain heart infusion supplemented with 0.75% Tween 80 and 2% glycerol. The alpha-toxin was purified by precipitation with polyethylene glycol 6000, followed by chromatography on Q-Sepharose, phenyl-agarose, and Mono-Q. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the toxin exhibited a single band with an M(r) of 200,000. The toxin also exhibited a single immunoprecipitin arc by crossed immunoelectrophoresis with antiserum against crude toxin. It was stable when stored at 4 degrees C and also following exposure to buffers with pHs in the range of 4 to 7. The toxin had a minimum lethal dose in mice of 5 to 10 ng, caused rounding of a variety of cells in tissue culture, and was negative in the rabbit ileal loop assay. The cytotoxic activity was inhibited by agents that affect receptor-mediated processes, and the toxin was less active on a CHO mutant cell line that is defective in endosomal acidification. The analysis of the amino acid composition revealed an unusually high proline content. The N-terminal sequence is Met-Leu-Ile-Thr-Arg-Glu-Gln-Leu-Met-Lys. Images PMID:8514395

  15. Cloning and sequencing of the genes for Shiga toxin from Shigella dysenteriae type 1.

    PubMed Central

    Strockbine, N A; Jackson, M P; Sung, L M; Holmes, R K; O'Brien, A D

    1988-01-01

    The structural genes for Shiga toxin, designated stx A and stx B, were cloned from Shigella dysenteriae type 1 3818T, and a nucleotide sequence analysis was performed. Both stx A and stx B were present on a single transcriptional unit, with stx A preceding stx B. The molecular weight calculated for the processed A subunit was 32,225, while the molecular weight of the processed B subunit was 7,691. Comparison of the nucleotide sequences for Shiga toxin and Shiga-like toxin I (SLT-I) from Escherichia coli revealed that the genes for Shiga toxin and SLT-I were greater than 99% homologous; three nucleotide changes were detected in three separate codons of the A subunits. Only one of these codon differences resulted in a change in the amino acid sequence: a threonine in Shiga toxin at position 45 of the A subunit compared with a serine in the corresponding position in SLT-I. Furthermore, Shiga toxin and SLT-I had identical signal peptides for the A and B subunits, as well as identical ribosome-binding sites, a putative promoter, and iron-regulated operator sequences. These findings indicate that Shiga and SLT-I are essentially the same toxin. Southern hybridization studies with total cellular DNA from several Shigella strains and internal toxin probes for SLT-I and its antigenic variant SLT-II showed that a single fragment in S. dysenteriae type 1 hybridized strongly with the internal SLT-I probe. Fragments with weaker homology to the SLT-I probe were detected in S. flexneri type 2a but no other shigellae. No homology between the Shiga-like toxin II (SLT-II) probe and any of the Shigella DNAs was detected. Whereas SLT-I and SLT-II are phage encoded, no phage could be induced from S. dysenteriae type 1 or other Shigella spp. tested. These results suggest that the Shiga (SLT-I) toxin genes responsible for high toxin production are present in a single copy in S. dysenteriae type 1 but not in other shigellae. The findings further suggest that SLT-II genes are absent in

  16. Comparison of the relative toxicities of Shiga-like toxins type I and type II for mice.

    PubMed Central

    Tesh, V L; Burris, J A; Owens, J W; Gordon, V M; Wadolkowski, E A; O'Brien, A D; Samuel, J E

    1993-01-01

    In earlier studies using a streptomycin-treated mouse model of infection caused by enterohemorrhagic Escherichia coli (EHEC), animals fed Shiga-like toxin type II (SLT-II)-producing strains developed acute renal cortical necrosis and died, while mice fed Shiga-like toxin type I (SLT-I)-producing clones did not die (E. A. Wadolkowski, L. M. Sung, J. A. Burris, J. E. Samuel, and A. D. O'Brien, Infect. Immun. 58:3959-3965, 1990). To examine the bases for the differences we noted between the two toxins in the murine infection model, we injected mice with purified toxins and carried out histopathological examinations. Despite the genetic and structural similarities between the two toxins, SLT-II had a 50% lethal dose (LD50) which was approximately 400 times lower than that of SLT-I when injected intravenously or intraperitoneally into mice. Histopathologic examination of toxin-injected mice revealed that detectable damage was limited to renal cortical tubule epithelial cells. Passive administration of anti-SLT-II antibodies protected mice from SLT-II-mediated kidney damage and death. Immunofluorescence staining of normal murine kidney sections incubated with purified SLT-I or SLT-II demonstrated that both toxins bound to cortical tubule and medullary duct epithelial cells. Compared with SLT-I, SLT-II was more heat and pH stable, suggesting that SLT-II is a relatively more stable macromolecule. Although both toxins bound to globotriaosylceramide, SLT-I bound with a higher affinity in a solid-phase binding assay. Differences in enzymatic activity between the two toxins were not detected. These data suggest that structural/functional differences between the two toxins, possibly involving holotoxin stability and/or receptor affinity, may contribute to the differential LD50s in mice. Images PMID:8335369

  17. [Exfoliative erythroderma and infective dermatitis in an infant infected with human T-lymphotropic virus type I (HTLV I)].

    PubMed

    Pérez C, Lilian; Villarroel B, Julia; Reyes J, Alejandra; Benavides M, Alicia; Muñoz O, Carla

    2007-04-01

    We report a HTLV-I positive infant, whose infection was confirmed by polymerase chain reaction. The infant presented with an acute, severe, generalized eczema, exfoliation and severe erythroderma that yielded to an acute proteic malnutrition and frequent staphylococcal infections, unresponsive to treatment, since the second month of life. Immunodeficiencies from other origin and other causes of erythroderma were ruled out. The histopathology studies and clinical course yielded to the diagnosis of infective dermatitis associated to HTLV-I. A review of the literature is performed.

  18. Type A and type B botulism in the North: first reported cases due to toxin other than type E in Alaskan Inuit.

    PubMed Central

    Barrett, D. H.; Eisenberg, M. S.; Bender, T. R.; Burks, J. M.; Hatheway, C. L.; Dowell, V. R.

    1977-01-01

    Botulism outbreaks shown to be due to type A and type B toxin occurred in Alaska, a region previously known for only type E botulism. The outbreak due to type A toxin involved three people, two of whom died. The outbreak due to type B toxin involved nine people, none of whom died. Both outbreaks were in Inuit villages, and native foods were incriminated. The occurrence of these outbreaks strongly suggests that Clostridium botulinum, types A and B are indigenous to Alaska. The outbreaks underscore the need for initial treatment of patients with antitoxin that is trivalent (ABE), even in Arctic regions. PMID:332309

  19. Development of antitoxin with each of two complementary fragments of Clostridium botulinum type B derivative toxin.

    PubMed Central

    Kozaki, S; Miyazaki, S; Sakaguchi, G

    1977-01-01

    Two fragments with molecular weights of 111,000 (fragment I) and 59,000 (fragment II) were separated from each other by gel filtration of dithiothreitol and urea-treated, trypsinized derivative toxin (molecular weight, 170,000) of the proteolytic Okra strain of Clostridium botulinum type B on a column of Sephadex G-200 (superfine) with a buffer containing dithiothreitol and urea. Upon removal of dithiothreitol and urea by dialysis, the two fragments reassembled to reconstruct the derivative toxin molecule. Both fragments were immunogenic, and both anti-fragments neutralized type B toxin. The neutralizing activities of both anti-fragment I and anti-fragment II were, however, lower than that of the anti-derivative toxin, suggesting that the molecular integrity of derivative toxin is essential for sufficient production of the neutralizing antibody. The immunological difference found between type B toxin from a proteolytic strain and that from a nonproteolytic strain was ascribed to the antigenic difference of fragment I. Images PMID:412790

  20. Development of antitoxin with each of two complementary fragments of Clostridium botulinum type B derivative toxin.

    PubMed

    Kozaki, S; Miyazaki, S; Sakaguchi, G

    1977-12-01

    Two fragments with molecular weights of 111,000 (fragment I) and 59,000 (fragment II) were separated from each other by gel filtration of dithiothreitol and urea-treated, trypsinized derivative toxin (molecular weight, 170,000) of the proteolytic Okra strain of Clostridium botulinum type B on a column of Sephadex G-200 (superfine) with a buffer containing dithiothreitol and urea. Upon removal of dithiothreitol and urea by dialysis, the two fragments reassembled to reconstruct the derivative toxin molecule. Both fragments were immunogenic, and both anti-fragments neutralized type B toxin. The neutralizing activities of both anti-fragment I and anti-fragment II were, however, lower than that of the anti-derivative toxin, suggesting that the molecular integrity of derivative toxin is essential for sufficient production of the neutralizing antibody. The immunological difference found between type B toxin from a proteolytic strain and that from a nonproteolytic strain was ascribed to the antigenic difference of fragment I.

  1. Thermally exfoliated graphite oxide

    NASA Technical Reports Server (NTRS)

    Prud'Homme, Robert K. (Inventor); Aksay, Ilhan A. (Inventor); Abdala, Ahmed (Inventor)

    2011-01-01

    A modified graphite oxide material contains a thermally exfoliated graphite oxide with a surface area of from about 300 sq m/g to 2600 sq m/g, wherein the thermally exfoliated graphite oxide displays no signature of the original graphite and/or graphite oxide, as determined by X-ray diffraction.

  2. Resistance of ovine, caprine and bovine endothelial cells to Clostridium perfringens type D epsilon toxin in vitro.

    PubMed

    Uzal, F A; Rolfe, B E; Smith, N J; Thomas, A C; Kelly, W R

    1999-08-01

    Ovine, caprine and bovine endothelial cells were grown in vitro and challenged with Clostridium perfringens type D epsilon toxin to compare their susceptibility to this toxin. Madin Darby canine kidney (MDCK) cells, which are known to be susceptible to epsilon toxin, were used as a positive control. No morphological alterations were observed in any of the endothelial cell cultures tested, even after challenging with doses as high as 1200 MLD50/ml of epsilon toxin. MDCK cells showed contour rounding and nuclear condensation as early as 30 min after exposure to 100 MLD50/ml of epsilon toxin and after 60 min of exposure to 12.5 MLD50/ml of the same toxin. All the MDCK cells were dead after 3 h of exposure to all concentrations of epsilon toxin. The results indicate that ovine, caprine and bovine endothelial cells are not morphologically responsive to the action of epsilon toxin in vitro.

  3. Both epsilon-toxin and beta-toxin are important for the lethal properties of Clostridium perfringens type B isolates in the mouse intravenous injection model.

    PubMed

    Fernandez-Miyakawa, Mariano E; Fisher, Derek J; Poon, Rachael; Sayeed, Sameera; Adams, Vicki; Rood, Julian I; McClane, Bruce A; Uzal, Francisco A

    2007-03-01

    Clostridium perfringens is capable of producing up to 15 toxins, including alpha-toxin (CPA), beta-toxin (CPB), epsilon-toxin (ETX), enterotoxin, beta2-toxin (CPB2), and perfringolysin O. Type B isolates, which must produce CPA, CPB, and ETX, are associated with animal illnesses characterized by sudden death or acute neurological signs, with or without intestinal damage. Type B pathogenesis in ruminants is poorly understood, with some animals showing lesions and clinical signs similar to those caused by either type C or type D infections. It is unknown whether host or environmental conditions are dominant for determining the outcome of type B disease or if disease outcomes are determined by variable characteristics of type B isolates. To help clarify this issue, 19 type B isolates were evaluated for toxin production during late-log-phase growth via quantitative Western blotting and by biological activity assays. Most type B isolates produced CPB levels similar to those produced by type C isolates in vitro and have the potential to produce genotype C-like disease. The lethality of type B isolate supernatants administered intravenously to mice was evaluated with or without prior trypsin treatment, and monoclonal antibody neutralization studies also were performed. Correlation analyses comparing toxin levels in type B supernatants versus lethality and neutralization studies both found that the main contributor to lethality without pretreatment with trypsin was CPB, whereas neutralization studies indicated that CPB and ETX were both important after trypsin pretreatment. At least part of the CPB produced by type B isolates remained active after trypsin treatment. However, the overall lethalities of most supernatants were lower after trypsin pretreatment. Also, there was a significant association between ETX, CPB2, and CPA production in vitro among type B isolates. However, our results suggest that both CPB and ETX are likely the most important contributors to the

  4. Botulinum toxin type A in the healing of chronic lesion following bilateral spasticity of gluteus muscle.

    PubMed

    Cigna, Emanuele; Maruccia, Michele; Fanelli, Benedetta; Scuderi, Nicolò

    2014-08-01

    Use of botulinum toxin is expanding as the clinical studies demonstrate new potential therapeutic applications. In rehabilitation, botulinum toxin is above all used as adjunct therapy for the treatment of spasticity, but it may prove useful for other atypical clinical situations. A 17-year-old man had a sub-arachnoid haemorrhage following the rupture of cerebral aneurism. The patient presented gluteus maximus and medius bilaterally spasticity that produced a chronic lesion in the intergluteal cleft, a flexed wrist and a flexed elbow. As treatment for this spasticity, a total of 100 U botulinum toxin type A were injected into the glutei muscles. This treatment allowed for application of topical medication and subsequently, chronic lesion healing. Botulinum toxin A may be an important therapeutic aid for clinicians faced with treating persistent pathological conditions caused by spasticity. © 2012 The Authors. International Wound Journal © 2012 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  5. Electron Microscopy of the Toxin and Hemagglutinin of Type A Clostridium botulinum

    PubMed Central

    Boroff, Daniel A.; Nyberg, Sverker; Höglund, Stefan

    1972-01-01

    Electron micrographs of the toxin and the hemagglutinin of type A Clostridium botulinum showed the toxin to be either round or disclike particles of 4 to 4.5 nm. These particles could also be seen as arranged in long strands or tubules of 9 nm in width. The hemagglutinin appeared as a crystalloid monolayer of stacked particles of 9 nm forming regularly arranged structures of 20 nm. Seen in cross section, these structures appeared as tubules with a lumen of 9 nm. The regularity of the angle of 83° to the long axis of the structure in which the individual particles were arranged suggested that the hemagglutinins formed a helix with sufficient space within its coil to admit the strands of the toxins. A model of the possible arrangement of the toxin and the hemagglutinin in the native state is proposed. Images PMID:4565051

  6. Pathogenesis of brain damage produced in sheep by Clostridium perfringens type D epsilon toxin: a review.

    PubMed

    Finnie, J W

    2003-04-01

    Microvascular endothelial damage by the epsilon toxin of Clostridium perfringens type D appears to be the fundamental cause of cerebral parenchymal injury and lesions occur in a seemingly dose- and time-dependent manner. Large doses of circulating toxin produce a severe, generalised, vasogenic cerebral oedema and an acute or peracute clinical course to death. With lower doses of toxin, or in partially immune sheep, focal necrosis, often bilaterally symmetrical, occurs in certain selectively vulnerable brain regions, which appear to become fewer as the toxin dose is reduced. These cases follow a more protracted clinical course, but death is the usual outcome. The precise pathogenesis of the focal brain damage found in subacutely intoxicated sheep is unresolved, but several possible mechanisms are discussed.

  7. Detection of Clostridium novyi type B alpha toxin by cell culture systems.

    PubMed

    Borrmann, E; Schulze, F

    1999-07-01

    Ten permanent cell lines were examined for their reaction to the Clostridium novyi alpha toxin. The action of the toxin was determined after 3 days by microscopic examination and the MTT assay. The alpha toxin exhibited the strongest effect on ESH-L cells rather than other cell lines. Vero and SFT-R cells reacted in a comparable way, but less sensitively. We were able to show that the cytopathic effect on the three types of cells was neutralised by the international standard for gas gangrene antitoxin (C. novyi) but in no case by heterologous antisera. Our results have shown that the three cell lines were specific indicators for the detection of the cytopathic effect of alpha toxin. The cytopathic effect can be measured reproducibly by the cell culture assay used. These results are suitable as the starting point for the development of the neutralisation test using cell cultures.

  8. Horizontal gene transfer of chromosomal Type II toxin-antitoxin systems of Escherichia coli.

    PubMed

    Ramisetty, Bhaskar Chandra Mohan; Santhosh, Ramachandran Sarojini

    2016-02-01

    Type II toxin-antitoxin systems (TAs) are small autoregulated bicistronic operons that encode a toxin protein with the potential to inhibit metabolic processes and an antitoxin protein to neutralize the toxin. Most of the bacterial genomes encode multiple TAs. However, the diversity and accumulation of TAs on bacterial genomes and its physiological implications are highly debated. Here we provide evidence that Escherichia coli chromosomal TAs (encoding RNase toxins) are 'acquired' DNA likely originated from heterologous DNA and are the smallest known autoregulated operons with the potential for horizontal propagation. Sequence analyses revealed that integration of TAs into the bacterial genome is unique and contributes to variations in the coding and/or regulatory regions of flanking host genome sequences. Plasmids and genomes encoding identical TAs of natural isolates are mutually exclusive. Chromosomal TAs might play significant roles in the evolution and ecology of bacteria by contributing to host genome variation and by moderation of plasmid maintenance.

  9. Structure, Evolution, and Functions of Bacterial Type III Toxin-Antitoxin Systems

    PubMed Central

    Goeders, Nathalie; Chai, Ray; Chen, Bihe; Day, Andrew; Salmond, George P. C.

    2016-01-01

    Toxin-antitoxin (TA) systems are small genetic modules that encode a toxin (that targets an essential cellular process) and an antitoxin that neutralises or suppresses the deleterious effect of the toxin. Based on the molecular nature of the toxin and antitoxin components, TA systems are categorised into different types. Type III TA systems, the focus of this review, are composed of a toxic endoribonuclease neutralised by a non-coding RNA antitoxin in a pseudoknotted configuration. Bioinformatic analysis shows that the Type III systems can be classified into subtypes. These TA systems were originally discovered through a phage resistance phenotype arising due to a process akin to an altruistic suicide; the phenomenon of abortive infection. Some Type III TA systems are bifunctional and can stabilise plasmids during vegetative growth and sporulation. Features particular to Type III systems are explored here, emphasising some of the characteristics of the RNA antitoxin and how these may affect the co-evolutionary relationship between toxins and cognate antitoxins in their quaternary structures. Finally, an updated analysis of the distribution and diversity of these systems are presented and discussed. PMID:27690100

  10. Patient considerations in the treatment of cervical dystonia: focus on botulinum toxin type A

    PubMed Central

    Mills, Reversa R; Pagan, Fernando L

    2015-01-01

    Cervical dystonia is the most common form of focal dystonia characterized by involuntary muscle contractions causing abnormal movements and posturing of the head and neck and is associated with significant pain. Botulinum toxin is considered first-line therapy in the treatment of pain and abnormal head posturing associated with cervical dystonia. There are currently three botulinum toxin type A neurotoxins and one botulinum type B neurotoxin commercially available and US Food and Drug Administration (FDA) labeled for the treatment of cervical dystonia. This review will focus on the efficacy, safety, and therapeutic use of botulinum type A neurotoxins in the treatment of cervical dystonia. We conclude with a discussion of factors influencing toxin selection including therapeutic effect, duration of effect, side effect profile, cost, and physician preference. PMID:26082621

  11. Dual toxin-producing strain of Clostridium botulinum type Bf isolated from a California patient with infant botulism.

    PubMed

    Barash, Jason R; Arnon, Stephen S

    2004-04-01

    A retrospective study of Clostridium botulinum strains isolated from patients from California with infant botulism identified the fourth known C. botulinum strain that produces both type B and type F botulinum toxins. This unique strain represented 0.12% of the California infant botulism case isolates from 1976 to 2003. The relative concentrations of type B and F toxins produced were temperature dependent.

  12. Design and synthesis of type-III mimetics of ShK toxin

    NASA Astrophysics Data System (ADS)

    Baell, Jonathan B.; Harvey, Andrew J.; Norton, Raymond S.

    2002-04-01

    ShK toxin is a structurally defined, 35-residue polypeptide which blocks the voltage-gated Kv1.3 potassium channel in T-lymphocytes and has been identified as a possible immunosuppressant. Our interest lies in the rational design and synthesis of type-III mimetics of protein and polypeptide structure and function. ShK toxin is a challenging target for mimetic design as its binding epitope consists of relatively weakly binding residues, some of which are discontinuous. We discuss here our investigations into the design and synthesis of 1st generation, small molecule mimetics of ShK toxin and highlight any principles relevant to the generic design of type-III mimetics of continuous and discontinuous binding epitopes. We complement our approach with attempted pharmacophore-based database mining.

  13. Clostridium botulinum type A progenitor toxin binds to Intestine-407 cells via N-acetyllactosamine moiety.

    PubMed

    Kojima, Shoudou; Eguchi, Hironobu; Ookawara, Tomomi; Fujiwara, Noriko; Yasuda, Jun; Nakagawa, Kazuhiko; Yamamura, Takehira; Suzuki, Keiichiro

    2005-06-03

    Botulism is a highly fatal disease caused by the botulinum progenitor toxin. In this study, the role of oligosaccharides for the binding of botulinum type A progenitor toxin (type A PTX) to human intestinal cells was investigated. The binding of type A PTX to Intestine-407 cells was inhibited by the addition of N-acetyllactosamine, lactose, and galactose. Treatment of Intestine-407 cells with neuraminidase led to a significant increase in the binding of type A PTX, while further digestion of cell surface oligosaccharides by beta-galactosidase and beta-N-acetylhexosaminidase decreased the binding. These results indicate that the N-acetyllactosamine moiety is responsible for the binding of type A PTX. These findings were further confirmed by a binding assay using synthesized oligosaccharides. Interestingly, sialylation or fucosylation of oligosaccharides inhibited the binding of type A PTX. These data suggest that the type A PTX binds to intestinal cells via cell surface N-acetyllactosamine moiety.

  14. Liquid-type Botulinum Toxin Type A in Adductor Spasmodic Dysphonia: A Prospective Pilot Study.

    PubMed

    Cha, Wonjae; Jang, Jeon Yeob; Wang, Soo-Geun; Kang, Ji-Heon; Jo, Min-Gyu

    2017-05-01

    Botulinum toxin (BTX) has been widely used to treat adductor spasmodic dysphonia (ADSD). Most commercially available forms of BTX require reconstitution before use, which may increase the risk of contamination and requires careful titration. Recently, a liquid-type BTX type A (BTX-A) has been developed, which should simplify the procedure and enhance its efficacy. Herein, we present a prospective pilot study to investigate the efficacy and safety of liquid-type BTX-A in the treatment of ADSD. Twenty-six consecutive liquid-type BTX-A injections were performed in 12 patients with ADSD. We included as a control group 34 consecutive patients with ADSD who had previously undergone 52 vocal fold injection procedures with freeze-dried-type BTX-A. All patients in both groups had improvement of symptoms related to ADSD and period of normal voice. Most patients experienced breathiness, and the onset time, the peak response time, and the duration of breathiness were similar in both groups. The duration of effect (days) was 96.96 ± 18.91 and 77.38 ± 18.97 in the freeze-dried-type and the liquid-type groups, and the duration of benefit (days) was 80.02 ± 18.24 and 62.69 ± 19.73 in the freeze-dried-type and the liquid-type groups. To compare the efficacy between the freeze-dried-type and the liquid-type BTX-A, the sessions of the unilateral vocal fold injection were included and were categorized as group A (1 ~ 2 units BTX-A) and group B (2 ~ 3 units BTX-A), according to the dose per vocal fold. There was no significant difference of effect time between freeze-dried-type and liquid-type BTX-A groups. No adverse events related to BTX or vocal fold injection were reported. Liquid-type BTX-A is safe and effective for the treatment of spasmodic dysphonia. With the advantages of simple preparation, storage, and reuse and animal protein-free constituents, liquid-type BTX-A may be a good option in the treatment of spasmodic dysphonia. Copyright © 2017 The

  15. Effect of Botulinum Toxin Type A on TGF-β/Smad Pathway Signaling: Implications for Silicone-Induced Capsule Formation.

    PubMed

    Kim, Sena; Ahn, Moonsang; Piao, Yibo; Ha, Yooseok; Choi, Dae-Kyoung; Yi, Min-Hee; Shin, Nara; Kim, Dong Woon; Oh, Sang-Ha

    2016-11-01

    One of the most serious complications of breast surgery using implants is capsular contracture. Several preventive treatments have been introduced; however, the mechanism of capsule formation has not been resolved completely. The authors previously identified negative effects of botulinum toxin type A on capsule formation, expression of transforming growth factor (TGF)-β1, and differentiation of fibroblasts into myofibroblasts. Thus, the authors investigated how to prevent capsule formation by using botulinum toxin type A, particularly by means of TGF-β1 signaling, in human fibroblasts. In vitro, cultured human fibroblasts were treated with TGF-β1 and/or botulinum toxin type A. Expression of collagen, matrix metalloproteinase, and Smad was examined by Western blotting. The activation of matrix metalloproteinase was observed by gelatin zymography. In vivo, the effect of botulinum toxin type A on the phosphorylation of Smad2 in silicone-induced capsule formation was evaluated by immunocytochemistry. In vitro, the phosphorylation of Smad2 was inhibited by botulinum toxin type A treatment. The expression levels of collagen types 1 and 3 were inhibited by botulinum toxin type A treatment, whereas those of matrix metalloproteinase-2 and matrix metalloproteinase-9 were enhanced. Gelatin zymography experiments confirmed enhanced matrix metalloproteinase-2 activity in collagen degradation. In vivo, botulinum toxin type A treatment reduced capsule thickness and Smad2 phosphorylation in silicone-induced capsules. This study suggests that botulinum toxin type A plays an important role in the inhibition of capsule formation through the TGF-β/Smad signaling pathway. Therapeutic, V.

  16. A PCR approach to determine the distribution of toxin genes in closely related Clostridium species: Clostridium botulinum type C and D neurotoxins and C2 toxin, and Clostridium novyi alpha toxin.

    PubMed

    Heffron, Ann; Poxton, Ian R

    2007-02-01

    The closely related Clostridium novyi and Clostridium botulinum types C and D are of current interest because of their association with serious infections in injecting drug users (C. novyi type A) and equine and feline dysautonomias (C. botulinum types C/D). The species are defined by the major toxins they produce: the alpha toxin of C. novyi, and the type C and D neurotoxins of C. botulinum (BoNT/C and BoNT/D). The other major toxin produced by this group, and previously thought to be restricted to the botulinum types, is the chromosomally encoded C2--a binary toxin consisting of two components, I and II. In the current study 44 of these clostridia from the authors' culture collection were investigated--most of which had been identified previously by conventional biochemical tests as 'C. novyi type A'. The aim was to check the distribution of toxin genes by PCR to see if the identities were consistent with the genes carried, and to ascertain if the C2 gene was only found in authentic C. botulinum strains. Several combinations of the species-defining genes and the two components of the C2 genes were detected. Only the authentic BoNT/C- and BoNT/D-positive C. botulinum strains and one of two non-neurotoxic variants of type C carried genes for both components of the C2 toxin. Of the remaining 40 C. novyi type A-like strains, the gene for the alpha toxin was found in 22, with 19 of these also possessing the gene for component I (16) or component II (3) but not both. In the alpha toxin-negative strains (22), both of the C2 genes were detected in 5 strains (3 C. botulinum), with component I in 11 strains and neither gene in 6 strains.

  17. [Treatment and indication of botulinum toxin type A for limb spasticity - can we break the 6 months barrier?].

    PubMed

    Miyashiro, Ai

    2012-01-01

    It is common knowledge that recovery of motor function is limited at 6 months after the onset of stroke. But there are some reports that motor functions are improved with using botulinum toxin type A for limb spasticity in the maintenance stage of stroke. Though it has been thought that botulinum toxin type A works in the peripheral nerves so far, Caleo showed botulinum toxin can affect the central nervous system. We suspected botulinum toxin type A affected the spinal cord directly following retrograde transynaptic transport from our experiments and his reports. So, we deduce the abnormal stretch reflex is made a modification by affecting the spinal cord, not only the injected muscle is relaxed, but also motor function is improved. Botulinum toxin type A shows sustained activity up to only 3 months, so we think we should use sufficient dose of botulinum toxin which may cause weakness. Rehabilitation with injected muscles contractions is important soon after botulinum toxin treatment, because botulinum toxin has a specific affinity to cleave certain proteins involved in the mechanism of acetylcholine exocytosis. The new botuslinum toxins type A which decrease the risk of production of antibodies and diffusion of noninjected muscles are under development.

  18. In situ detection of the Clostridium botulinum type C1 toxin gene in wetland sediments with a nested PCR assay

    USGS Publications Warehouse

    Williamson, J.L.; Rocke, T.E.; Aiken, Judd M.

    1999-01-01

    A nested PCR was developed for detection of the Clostridium botulinum type C1 toxin gene in sediments collected from wetlands where avian botulism outbreaks had or had not occurred. The C1 toxin gene was detected in 16 of 18 sites, demonstrating both the ubiquitous distribution of C. botulinum type C in wetland sediments and the sensitivity of the detection assay.

  19. In Situ Detection of the Clostridium botulinum Type C1 Toxin Gene in Wetland Sediments with a Nested PCR Assay

    PubMed Central

    Williamson, Judy L.; Rocke, Tonie E.; Aiken, Judd M.

    1999-01-01

    A nested PCR was developed for detection of the Clostridium botulinum type C1 toxin gene in sediments collected from wetlands where avian botulism outbreaks had or had not occurred. The C1 toxin gene was detected in 16 of 18 sites, demonstrating both the ubiquitous distribution of C. botulinum type C in wetland sediments and the sensitivity of the detection assay. PMID:10388729

  20. Detecting and distinguishing among type 1 and type 2 Shiga toxins in human serum

    USDA-ARS?s Scientific Manuscript database

    Shiga toxins, also known as verotoxins, are a major virulence factor associated with Shiga toxin producing Escherichia coli (STEC). STEC are the E. coli responsible for many of the serious foodborne outbreaks of disease. We have developed a sensitive and specific mass spectrometry-based method of de...

  1. Elevation of the Corner of the Mouth Using Botulinum Toxin Type A

    PubMed Central

    Goldman, Alberto; Wollina, Uwe

    2010-01-01

    Indications for botulinum toxin type A have been constantly evolving, and it can currently be used in virtually any area of the face and neck. The authors present their experience with this neurotoxin in treating the platysmal bands and depressor anguli oris muscle with the purpose of cosmetically improving the anterior neck and lifting the oral commissure. PMID:21430826

  2. Employment of higher doses of botulinum toxin type A to reduce spasticity after stroke.

    PubMed

    Santamato, Andrea; Micello, Maria Francesca; Ranieri, Maurizio; Valeno, Giovanni; Albano, Antonio; Baricich, Alessio; Cisari, Carlo; Intiso, Domenico; Pilotto, Alberto; Logroscino, Giancarlo; Panza, Francesco

    2015-03-15

    Spasticity is a common disabling symptom for several neurological conditions. Botulinum toxin type A injection represents the gold standard treatment for focal spasticity with efficacy, reversibility, and low prevalence of complications. Current guidelines suggest a dose up to 600 units (U) of onabotulinumtoxinA/incobotulinumtoxinA or up to 1,500 U of abobotulinumtoxinA to treat post-stroke spasticity to avoid important adverse effects. However, recently, higher doses of botulinum toxin type A were employed, especially in case of upper and lower limb severe spasticity. With searches of US National Library of Medicine databases, we identified all studies published from December 1989 to July 2014 concerning the use of higher doses of this neurotoxin for spasticity treatment with at least a dose of 600 U of onabotulinumtoxinA and incobotulinumtoxinA or 1,800 U of abobotulinumtoxinA. The cumulative body of evidence coming from the eight studies selected suggested that higher doses of botulinum toxin type A appeared to be efficacious in reducing spasticity of the upper and lower limbs after stroke, with adverse effects generally mild. However, further investigations are needed to determine the safety and reproducibility in larger case series or randomized clinical trials of higher doses of botulinum toxin type A also after repeated injections. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. The use of botulinum toxin type A treatment in children with spasticity.

    PubMed

    Sarioglu, Berrak; Serdaroglu, Gul; Tutuncuoglu, Sarenur; Ozer, Esra A

    2003-10-01

    The current modalities in managing spastic children have some limitations; thus, alternative therapeutic agents are in need. The purpose of this study is to investigate whether intramuscular botulinum toxin type A administration may be an alternative agent in the treatment of children with cerebral palsy. Eighteen children who were aged between 3 and 17 years and manifested cerebral palsy were administered intramuscular botulinum toxin type A with a total dose of 6 U/kg body weight. Outcome measurements were determined with four methods, including Ashworth Spasticity Scale, standardized videotape assessments, observational gait analysis, and walking velocity. Ashworth Spasticity Scale and videotape assessments were statistically significant before and after treatment in all muscles (P < 0.001). The best improvement in video gait analysis was evident at week 8. The botulinum toxin type A injections yielded an improved walking velocity at all visits. The observational gait analysis and walking velocity demonstrated an improvement after treatment in the gastrocnemius-injected group (P < 0.001). In conclusion, intramuscular botulinum toxin type A administration may be effective in children with cerebral palsy, especially at week 4 and when injected in gastrocnemius.

  4. Detection of botulinum toxin types A, B, E, and F activity using the quail embryo

    USDA-ARS?s Scientific Manuscript database

    We recently demonstrated an effective new model for the detection of botulinum toxin type A using quail embryos in place of the mouse model. These experiments demonstrated that the Japanese quail embryo at 15 days of incubation was an effective vertebrate animal model to detect the activity of botu...

  5. Comparative study of biological activity of four botulinum toxin type A preparations in mice.

    PubMed

    Chung, Myung Eun; Song, Dae Heon; Park, Joo Hyun

    2013-01-01

    Units of available botulinum toxin preparations are not interchangeable, and the dose-conversion ratios between such preparations remain controversial. To compare the efficacy and safety of four botulinum toxin type A preparations. Murine gastrocnemius compound muscle action potentials (CMAPs) were recorded before and after injecting the four botulinum toxin preparations (onabotulinumtoxinA, abobotulinumtoxinA, new botulinum toxin, and incobotulinumtoxinA). In all preparations, CMAP amplitudes decreased until 4 days after receiving the injection and then gradually recovered. On postinjection day 84, the amplitudes returned to baseline in all groups except the high-dose groups. CMAP amplitude in the contralateral limb also decreased up to postinjection days 4 to 7 and then gradually returned to baseline by postinjection day 28. The dose-conversion ratio between onabotulinumtoxinA and abobotulinumtoxinA was determined to be 1:2.6; previous reports of 1:3 were considered too high. A dose-conversion ratio between onabotulinumtoxinA and new botulinum toxin of 1:1 was deemed appropriate. OnabotulinumtoxinA and incobotulinumtoxinA demonstrated a dose-conversion ratio of 1:1.07. The efficacy of incobotulinumtoxinA was slightly lower than that of onabotulinumtoxinA. These dose-conversion ratios are applicable solely from an efficacy standpoint and not for safety. This study was conducted in mice, so it may not translate perfectly to human applications. © 2012 by the American Society for Dermatologic Surgery, Inc. Published by Wiley Periodicals, Inc.

  6. Pharmacological and biochemical studies of cytotoxicity of Clostridium novyi type A alpha-toxin.

    PubMed Central

    Bette, P; Frevert, J; Mauler, F; Suttorp, N; Habermann, E

    1989-01-01

    The actions of apparently homogeneous alpha-toxin from Clostridium novyi type A were studied in order to develop an in vitro system which closely mimics its in vivo effects and to search for the mode of poisoning. Time to death (by intravenous injection of mice) was inversely related to dose, with a detection limit of about 200 ng/kg of body weight at 100 h. Injections of 2.5 ng or more into the rat paw led to a slowly (maximum after about 30 h) developing, dose-dependent edema which was useful as a quantitative in vivo assay based on volumetry. Vascular leakage was due to gap formation between endothelial cells. Similarly, endothelial cells cultured from pig pulmonary artery lost their "cobblestone" arrangement after a dose-dependent lag period of some hours after poisoning. The morphological changes were accompanied by depression of uptake or incorporation of [3H]uridine. A quantitative in vitro assay was established on the inhibition of [3H]uridine incorporation. As in animals, the action of alpha-toxin started with a few nanograms per milliliter and proceeded slowly for at least 1 day but became resistant to antitoxin within 2 h of exposure. The toxin action is not limited to endothelial cells, since chicken embryonic cells, a mouse fibroblast line (L-929), and a rat phaeochromocytoma line (PC-12) behaved similarly. Alpha-toxin was found to differ from other bacterial toxins investigated whose modes of action are already known. Images PMID:2744858

  7. Effect of fermentation conditions on toxin production by Clostridium botulinum type B.

    PubMed

    Siegel, L S; Metzger, J F

    1980-12-01

    To obtain high yields of toxin for the preparation of purified neurotoxoids, we examined the time of appearance and the quantity of toxin produced by the Bean strain of Clostridium botulinum type B under various conditions by using a fermentor system. The medium employed consisted of 2.0% casein hydrolylsate and 1.5% yeast extract plus an appropriate concentration of glucose. The maximum toxin concentration (4 x 10(5) to 5 x 10(5) mouse median lethal doses per ml) was attained within 48 h under the following fermentation conditions: an initial glucose concentration of 0.5 or 1.0%, a temperature of 35 degrees C, a nitrogen overlay at a rate of 5 liters/min, and an agitation rate of 50 rpm.

  8. Failure to Inhibit In Vitro or In Vivo Acetycholinesterase with Botulinum Toxin Type A

    PubMed Central

    Simpson, Lance L.; Morimoto, Hiromi

    1969-01-01

    An attempt has been made to replicate an earlier finding that type A botulinum toxin can inhibit the in vitro activity of acetylcholinesterase. Two methods of enzyme assay were employed, but with neither technique were we able to reproduce the finding of in vitro enzyme inhibition. In fact, an examination of the data from the previous report leads us to question the possibility of the observations that were given. Furthermore, an investigation was carried out to determine if botulinum toxin can exert an inhibiting effect on acetylcholinesterase that is situated in the biological tissue. The answer again is negative. The experimental observations, coupled with several mathematical computations, do not support the notion that botulinum toxin is an acetylcholinesterase inhibitor. PMID:5773011

  9. Why so narrow: Distribution of anti-sense regulated, type I toxin-antitoxin systems compared with type II and type III systems

    PubMed Central

    Coray, Dorien S.; Heinemann, Jack A.; Gardner, Paul P.

    2017-01-01

    ABSTRACT Toxin-antitoxin (TA) systems are gene modules that appear to be horizontally mobile across a wide range of prokaryotes. It has been proposed that type I TA systems, with an antisense RNA-antitoxin, are less mobile than other TAs that rely on direct toxin-antitoxin binding but no direct comparisons have been made. We searched for type I, II and III toxin families using iterative searches with profile hidden Markov models across phyla and replicons. The distribution of type I toxin families were comparatively narrow, but these patterns weakened with recently discovered families. We discuss how the function and phenotypes of TA systems as well as biases in our search methods may account for differences in their distribution. PMID:28067598

  10. Positive botulinum toxin type a response is a prognosticator for migraine surgery success.

    PubMed

    Lee, Michelle; Monson, Mikhal A; Liu, Mengyuan T; Reed, Deborah; Guyuron, Bahman

    2013-04-01

    The objective of the study was to determine whether botulinum toxin type A injections can serve as a prognosticator for migraine surgery success. Patients who underwent migraine surgery from 2000 to 2010 by the senior author (B.G.) were reviewed. Patients were included if they had botulinum toxin type A injection before surgery; had completed postinjection, postsurgery Migraine Headache Questionnaires; and had at least 1-year follow-up. Outcome variables include patient demographics and Migraine Headache Index. Treatment success was defined as at least a 50 percent reduction in Migraine Headache Index. One hundred eighty-eight patients were included; 144 reported successful migraine headache reduction after injection (success group) and 44 did not (failure group). The groups were well matched for age, migraine headache characteristics, and number of surgical sites (p > 0.05). The surgery success rate was significantly higher in the success group overall (90.3 percent versus 72.3, p = 0.003), and in patients who reported botulinum toxin type A success and subsequent same-site surgery (97.9 percent versus 71.4 percent, p < 0.0001). Botulinum toxin type A success was prognostic for surgery success at the frontal trigger site (trigger site I) (92.5 percent versus 69.2 percent, p = 0.012), the temporal trigger site (trigger site II) (95.5 percent versus 73.3 percent, p = 0.005), and the occipital trigger site (trigger site IV) (95.9 percent versus 62.5 percent, p = 0.0003). Six patients had exclusively septum or turbinate (site III) surgery, and all failed injections. Positive botulinum toxin type A response is a significant predictor of migraine surgery success. When injections fail, nonmuscular abnormalities should be considered.

  11. Keeping the Wolves at Bay: Antitoxins of Prokaryotic Type II Toxin-Antitoxin Systems.

    PubMed

    Chan, Wai Ting; Espinosa, Manuel; Yeo, Chew Chieng

    2016-01-01

    In their initial stages of discovery, prokaryotic toxin-antitoxin (TA) systems were confined to bacterial plasmids where they function to mediate the maintenance and stability of usually low- to medium-copy number plasmids through the post-segregational killing of any plasmid-free daughter cells that developed. Their eventual discovery as nearly ubiquitous and repetitive elements in bacterial chromosomes led to a wealth of knowledge and scientific debate as to their diversity and functionality in the prokaryotic lifestyle. Currently categorized into six different types designated types I-VI, type II TA systems are the best characterized. These generally comprised of two genes encoding a proteic toxin and its corresponding proteic antitoxin, respectively. Under normal growth conditions, the stable toxin is prevented from exerting its lethal effect through tight binding with the less stable antitoxin partner, forming a non-lethal TA protein complex. Besides binding with its cognate toxin, the antitoxin also plays a role in regulating the expression of the type II TA operon by binding to the operator site, thereby repressing transcription from the TA promoter. In most cases, full repression is observed in the presence of the TA complex as binding of the toxin enhances the DNA binding capability of the antitoxin. TA systems have been implicated in a gamut of prokaryotic cellular functions such as being mediators of programmed cell death as well as persistence or dormancy, biofilm formation, as defensive weapons against bacteriophage infections and as virulence factors in pathogenic bacteria. It is thus apparent that these antitoxins, as DNA-binding proteins, play an essential role in modulating the prokaryotic lifestyle whilst at the same time preventing the lethal action of the toxins under normal growth conditions, i.e., keeping the proverbial wolves at bay. In this review, we will cover the diversity and characteristics of various type II TA antitoxins. We shall

  12. Keeping the Wolves at Bay: Antitoxins of Prokaryotic Type II Toxin-Antitoxin Systems

    PubMed Central

    Chan, Wai Ting; Espinosa, Manuel; Yeo, Chew Chieng

    2016-01-01

    In their initial stages of discovery, prokaryotic toxin-antitoxin (TA) systems were confined to bacterial plasmids where they function to mediate the maintenance and stability of usually low- to medium-copy number plasmids through the post-segregational killing of any plasmid-free daughter cells that developed. Their eventual discovery as nearly ubiquitous and repetitive elements in bacterial chromosomes led to a wealth of knowledge and scientific debate as to their diversity and functionality in the prokaryotic lifestyle. Currently categorized into six different types designated types I–VI, type II TA systems are the best characterized. These generally comprised of two genes encoding a proteic toxin and its corresponding proteic antitoxin, respectively. Under normal growth conditions, the stable toxin is prevented from exerting its lethal effect through tight binding with the less stable antitoxin partner, forming a non-lethal TA protein complex. Besides binding with its cognate toxin, the antitoxin also plays a role in regulating the expression of the type II TA operon by binding to the operator site, thereby repressing transcription from the TA promoter. In most cases, full repression is observed in the presence of the TA complex as binding of the toxin enhances the DNA binding capability of the antitoxin. TA systems have been implicated in a gamut of prokaryotic cellular functions such as being mediators of programmed cell death as well as persistence or dormancy, biofilm formation, as defensive weapons against bacteriophage infections and as virulence factors in pathogenic bacteria. It is thus apparent that these antitoxins, as DNA-binding proteins, play an essential role in modulating the prokaryotic lifestyle whilst at the same time preventing the lethal action of the toxins under normal growth conditions, i.e., keeping the proverbial wolves at bay. In this review, we will cover the diversity and characteristics of various type II TA antitoxins. We shall

  13. Toxin Transport by A-B Type of Toxins in Eukaryotic Target Cells and Its Inhibition by Positively Charged Heterocyclic Molecules.

    PubMed

    Benz, Roland; Barth, Holger

    2017-07-04

    A-B types of toxins are among the most potent bacterial protein toxins produced by gram-positive bacteria. Prominent examples are the tripartite anthrax toxin of Bacillus anthracis and the different A-B type clostridial toxins that are the causative agents of severe human and animal diseases and could serve as biological weapons. The components of all these toxins comprise one binding/transport (B) subunit and one or two separate, non-linked enzymatically active (A) subunits. The A and B subunits are separately produced and secreted by the pathogenic gram-positive bacteria and must assemble on the surface of eukaryotic target cells to form biologically active toxin complexes. The B components are cleaved by proteases to generate the biologically active species that binds to receptors on the surface of the target cells and form there oligomers which bind the A subunits. The AB complexes are internalized by receptor-mediated endocytosis and reach early or late endosomes that become acidified. Subsequently, the B components form channels in endosomal membranes that are indispensable for the transport of the enzymatic subunits across these membranes into the cytosol of target cells via the trans-membrane channels. In addition to the channels formed by the B components, host cell factors including chaperones and further folding helper enzymes are involved in the import of the enzymatic subunits into the cytosol of eukaryotic cells. Positively charged heterocyclic molecules, such as chloroquine and related aminoquinolinium and azolopyridinium salts have been shown in recent years to bind with high affinity to the channels formed by the B components of binary toxins. Since binding to the B components is also a prerequisite for transport of the A components across the endosomal membranes the channel blockers also prevent transport of the A subunits into the host cell cytosol. The inhibition of toxin uptake into cells by such pharmacological compounds should also be of

  14. The Mechanism of the Beneficial Effect of Botulinum Toxin Type a Used in the Treatment of Temporomandibular Joints Dysfunction.

    PubMed

    Malgorzata, Pihut; Piotr, Ceranowicz; Edward, Kijak

    2017-01-01

    In the course of temporomandibular joint, dysfunctions very often occur to the excessive increase in tension of masticatory muscles, so the main aim of the treatment is reduction of this hypertension of muscles. For this reason, we use botulinum toxin type A, which is produced by Grampositive Clostridium bacteria. There are six serotypes of the toxin: A, B, C, D, E, F, and G. The botulinum toxin type A was first isolated in 1920s. Today, botulinum toxin type A is used increasingly more often as an efficient and patient-friendly therapy in neurology, ophthalmology, neurology, urology and laryngology. The aim of the article was to review the literature and description of the current knowledge concerned with mechanism of action of botulinum toxin type A, clinical applications and metabolic determinants of muscle contraction and the beneficial effect of this drug on the state of muscle tension. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  15. The early effects of Clostridium perfringens type D epsilon toxin in ligated intestinal loops of goats and sheep.

    PubMed

    Fernandez Miyakawa, M E; Uzal, F A

    2003-04-01

    Clostridium perfringens type D produces enterotoxaemia in goats, sheep and other animals. The disease is caused by C. perfringens epsilon toxin and, while enterotoxaemia in goats is usually characterized by enterocolitis, the disease in sheep is characterized by systemic lesions (such as lung and brain oedema) with minor and inconsistent changes observed in the intestine. A possible explanation for these differences is that epsilon toxin is more promptly absorbed by the ovine than by the caprine intestine. In an attempt to clarify this, we examined the early effects of epsilon toxin on caprine and ovine intestine. Intestinal loop assays were performed to analyse the physiological and morphological changes induced by epsilon toxin in the intestine of these species. Fluid accumulation was observed in caprine and ovine ileum and colon treated with epsilon toxin. Ileal loops from goats treated with epsilon toxin retained sodium and water earlier than ovine ileal loops treated with the same toxin. Histological analysis showed morphological alterations in the colon of both species as early as 2 h after the commencement of epsilon toxin treatment: these changes were more marked in goats than in sheep. No morphological changes were observed in the ileum of either species after 4 h incubation with epsilon toxin. These results suggest that epsilon toxin modifies ion and water transport in the small and the large intestine of goats and sheep through different mechanisms.

  16. Growth conditions of clostridium perfringens type B for production of toxins used to obtain veterinary vaccines.

    PubMed

    Viana Brandi, Igor; Domenici Mozzer, Otto; Jorge, Edson Vander; Vieira Passos, Frederico Jose; Lopes Passos, Flavia Maria; Cangussu, Alex Sander Rodrigues; Macedo Sobrinho, Eliane

    2014-09-01

    The diseases caused for Clostridium perfringens are generically called enterotoxemias because toxins produced in the intestine may be absorbed into the general circulation. C. perfringens type B, grown in batch fermentation, produced toxins used to obtain veterinary vaccines. Glucose in concentrations of 1.4-111.1 mM was used to define the culture medium. The minimum concentration for a satisfactory production of vaccines against clostridial diseases was 55.6 mM. Best results were brought forth by meat and casein peptones, both in the concentration 5.0 g l(-1) in combination with glucose and a culture pH maintained at 6.5 throughout the fermentation process. The production of lactic, acetic and propionic organic acids was observed. Ethanol was the metabolite produced in the highest concentration when cultures maintained steady pH of 6.5 with exception of cultures with initial glucose concentration of 1.4 mM, where the highest production was of propionic acid. Maximal cell concentration and the highest toxin title concomitantly low yield coefficient to organic acids and ethanol were obtained using basal medium containing 111.1 mM glucose under a controlled pH culture (pH) 6.5 in batch fermentations of C. perfringens type B. These data contribute to improve process for industrial toxin production allowing better condition to produce a toxoid vaccine.

  17. Type II Toxin-Antitoxin Systems in the Unicellular Cyanobacterium Synechocystis sp. PCC 6803.

    PubMed

    Kopfmann, Stefan; Roesch, Stefanie K; Hess, Wolfgang R

    2016-07-21

    Bacterial toxin-antitoxin (TA) systems are genetic elements, which are encoded by plasmid as well as chromosomal loci. They mediate plasmid and genomic island maintenance through post-segregational killing mechanisms but may also have milder effects, acting as mobile stress response systems that help certain cells of a population in persisting adverse growth conditions. Very few cyanobacterial TA system have been characterized thus far. In this work, we focus on the cyanobacterium Synechocystis 6803, a widely used model organism. We expand the number of putative Type II TA systems from 36 to 69 plus seven stand-alone components. Forty-seven TA pairs are located on the chromosome and 22 are plasmid-located. Different types of toxins are associated with various antitoxins in a mix and match principle. According to protein domains and experimental data, 81% of all toxins in Synechocystis 6803 likely exhibit RNase activity, suggesting extensive potential for toxicity-related RNA degradation and toxin-mediated transcriptome remodeling. Of particular interest is the Ssr8013-Slr8014 system encoded on plasmid pSYSG, which is part of a larger defense island or the pSYSX system Slr6056-Slr6057, which is linked to a bacterial ubiquitin-like system. Consequently, Synechocystis 6803 is one of the most prolific sources of new information about these genetic elements.

  18. The receptor and transporter for internalization of Clostridium botulinum type C progenitor toxin into HT-29 cells.

    PubMed

    Nishikawa, Atsushi; Uotsu, Nobuo; Arimitsu, Hideyuki; Lee, Jae-Chul; Miura, Yutaka; Fujinaga, Yukako; Nakada, Hiroshi; Watanabe, Toshihiro; Ohyama, Tohru; Sakano, Yoshiyuki; Oguma, Keiji

    2004-06-25

    Orally ingested botulinum toxin enters the circulatory system and eventually reaches the peripheral nerves, where it elicits a response of neurological dysfunction. In this study, we report the important findings concerning the mechanism of Clostridium botulinum type C progenitor toxin (C16S) endocytic mechanism. C16S toxin bound to high molecular weight proteins on the surface of human colon carcinoma HT-29 cells and was internalized, but not if the cells were pretreated with neuraminidase. Benzyl-GalNAc which inhibited O-glycosylation of glycoproteins also interfered in the toxin's ability to bind the cell surface. On the other hand, the toxin was internalized in spite of pretreatment of the cells with PPMP, an inhibitor of ganglioside synthesis. These results suggest that the glycoproteins, like mucin, fulfill the important roles of receptor and transporter of C16S toxin.

  19. Use of immunofluorescence and animal tests to detect growth and toxin production by Clostridum botulinum type E in food.

    PubMed

    Midura, T; Taclindo, C; Nygaard, G S; Bodily, H L; Wood, R M

    1968-01-01

    The appearance of Clostridium botulinum type E organisms and of toxin in experimentally inoculated packages of turkey roll was followed to study the time relationship between the presence of vegetative cells and the demonstration of toxin. The presence of vegetative cells was determined by immunofluorescence, and animal tests were used to assay toxin production. Growth initiated from detoxified spores of C. botulinum type E resulted in toxin formation within 24 hr. Presence of fluorescing vegetative cells and of toxin coincided from 1 to 14 days of incubation. Beginning with the next testing date, day 21, differences were observed. Toxin could be detected for a longer time than vegetative cells. Neither toxin nor organisms could be found after 56 days of incubation. The mouse lethal dose tests (MLD per gram of turkey roll) showed fluctuations in the amount of toxin present throughout the period of testing. Maximal amounts of toxin were present during the period when fluorescing organisms were also more numerous. The applications of immunofluorescence in the study and in the diagnosis of botulism is discussed.

  20. Contact with enterocyte-like Caco-2 cells induces rapid upregulation of toxin production by Clostridium perfringens type C isolates

    PubMed Central

    Vidal, Jorge E.; Ohtani, Kaori; Shimizu, Tohru; McClane, Bruce A.

    2009-01-01

    Clostridium perfringens type C isolates cause necrotizing enteritis in humans and domestic animals. In vitro, type C isolates often produce beta toxin (CPB), beta2 toxin (CPB2), alpha toxin (CPA), perfringolysin O (PFO), and TpeL during (or after) late log-phase growth. In contrast, the current study found that many type C isolates respond to close contact with enterocyte-like Caco-2 cells by producing all toxins, except TpeL, much more rapidly than occurs during in vitro growth. This in vivo effect involves rapid transcriptional upregulation of the cpb, cpb2, pfoA and plc toxin genes. Rapid Caco-2 cell-induced upregulation of CPB and PFO production involves the VirS/VirR two-component system, since upregulated in vivo transcription of the pfoA and cpb genes was blocked by inactivating the virR gene and was reversible by complementation to restore VirR expression. However, the luxS quorum sensing system is not required for the rapid upregulation of type C toxin production induced by contact with Caco-2 cells. These results provide the first indication of host cell:pathogen cross-talk affecting toxin production kinetics by any pathogenic Clostridium spp., identify in vivo vs. in vitro differences in C. perfringens toxin expression, and implicate VirS/VirR as a possible contributor to some C. perfringens enteric diseases. PMID:19438515

  1. Botulinum toxin type A for aging face and aesthetic uses.

    PubMed

    Hexsel, Camile; Hexsel, Doris; Porto, Manoela Donida; Schilling, Juliana; Siega, Carolina

    2011-01-01

    Botulinum neurotoxin type A injection to correct and/or reverse the physical effects of aging process has become one of the most frequently requested cosmetic procedures at an outpatient setting. Careful clinical evaluation together with proper use of the techniques, including pre- and post-procedures recommendations, reconstitution of the products, techniques, and doses, are described in this article. This article also covers the main indications of botulinum neurotoxin type A for aging face and other aesthetic uses, as well as some possible adverse reactions and their management. © 2011 Wiley Periodicals, Inc.

  2. Liquid phase exfoliated graphene for electronic applications

    NASA Astrophysics Data System (ADS)

    Sukumaran, Sheena S.; Jinesh, K. B.; Gopchandran, K. G.

    2017-09-01

    Graphene dispersions were prepared using the liquid phase exfoliation method with three different surfactants. One surfactant was used from each of the surfactant types, anionic, cationic, and non-ionic; those used, were sodium dodecylbenzene sulfonate (SDBS), cetyltrimethylammonium bromide (CTAB) and polyvinylpyrrolidone (PVP), respectively. Raman spectroscopy was used to investigate the number of layers and the nature of any defects present in the exfoliated graphene. The yield of graphene was found to be less with the non-ionic surfactant, PVP. The deconvolution of 2D peaks at ~2700 cm‑1 indicated that graphene prepared using these surfactants resulted in sheets consisting of few-layer graphene. The ratio of intensity of the D and G bands in the Raman spectra showed that edge defect density is high for samples prepared with SDBS compared to the other two, and is attributed to the smaller size of the graphene sheets, as shown in the electron micrographs. In the case of the dispersion in PVP, it is found that the sizes of the graphene sheets are highly sensitive to the concentration of the surfactant used. Here, we have made an attempt to investigate the local density of states in the graphene sheets by measuring the tunnelling current–voltage characteristics. Graphene layers have shown consistent p-type behaviour when exfoliated with SDBS and n-type behaviour when exfoliated with CTAB, with a larger band gap for graphene exfoliated using CTAB. Hence, in addition to the known advantages of liquid phase exfoliation, we found that by selecting suitable surfactants, to a certain extent it is possible to tune the band gap and determine the type of majority carriers.

  3. Dual Toxin-Producing Strain of Clostridium botulinum Type Bf Isolated from a California Patient with Infant Botulism

    PubMed Central

    Barash, Jason R.; Arnon, Stephen S.

    2004-01-01

    A retrospective study of Clostridium botulinum strains isolated from patients from California with infant botulism identified the fourth known C. botulinum strain that produces both type B and type F botulinum toxins. This unique strain represented 0.12% of the California infant botulism case isolates from 1976 to 2003. The relative concentrations of type B and F toxins produced were temperature dependent. PMID:15071029

  4. Use of botulinum toxin type A in the management of patients with neurological disorders: a national survey

    PubMed Central

    Smania, Nicola; Colosimo, Carlo; Bentivoglio, Anna Rita; Sandrini, Giorgio; Picelli, Alessandro

    2013-01-01

    Summary The aim of this survey was to provide an overview of important issues relating to therapeutic strategies based on botulinum toxin type A injection for the treatment of patients with neurological disorders. Two hundred and ten physicians from neurology and neurorehabilitation units in Italian hospitals answered a questionnaire exploring some clinical aspects of the use of botulinum toxin type A in patients with spasticity/dystonia. 66% of the physicians treated patients with dystonia, 80% treated adults with spasticity, and 35% treated children with cerebral palsy. Palpation with no instrumental guidance was the injection technique most commonly used for treating patients with dystonia, spasticity and cerebral palsy; 57% of the physicians evaluated patients instrumentally before toxin injection, while 45% assessed post-injection improvements by instrumental means; 78% of the physicians prescribed (when appropriate) rehabilitation procedures after toxin injection. Our results seem to show that the routine use of botulinum toxin in clinics is far from standardized. PMID:24598392

  5. Management of gummy smile with Botulinum Toxin Type-A: A case report

    PubMed Central

    Dinker, Sudeeptha; Anitha, A; Sorake, Abhinay; Kumar, Kishore

    2014-01-01

    A 23 year old female patient presented with the chief complaint of gummy smile after previously undergoing Orthodontic treatment. Patient had a straight profile with competent lips and during posed and unposed smile the patient exhibited excessive gingival display. Since the patient was unwilling to undergo Orthodontic treatment and apprehensive about surgical procedures, this problem was addressed by injecting Botulinum toxin type-A as an alternative treatment approach. Two weeks post treatment; on follow up examination, improved results were seen without any side effects. As a result, an attractive and confident smile was perceived by the patient. How to cite the article: Dinker S, Anitha A, Sorake A, Kumar K. Management of gummy smile with Botulinum Toxin Type-A: A case report. J Int Oral Health 2014;6(1):111-5. PMID:24653614

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

    PubMed

    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.

  7. Management of exfoliative glaucoma: challenges and solutions

    PubMed Central

    Holló, Gábor; Katsanos, Andreas; Konstas, Anastasios GP

    2015-01-01

    Exfoliative glaucoma is the most common type of secondary open-angle glaucoma worldwide. It is characterized by high intraocular pressure (IOP) and worse 24-hour IOP characteristics. In order to minimize progression, treatment of exfoliative glaucoma has to provide a low long-term mean IOP and good 24-hour IOP control. To achieve these goals, fixed-dose combination eye drops, argon and selective laser trabeculoplasty, and various forms of surgery (trabeculectomy, deep sclerectomy, viscocanalostomy, ab interno trabeculotomy, trabecular aspiration, and cataract surgery) all need to be considered during the long-term management of the disease. Since exfoliative glaucoma is a disease of the elderly, and is frequently associated with systemic vascular disease, interdisciplinary consultations are of great clinical importance. These management aspects and the current medical, laser, and surgical results are covered in this review, with a special focus on the needs of the general ophthalmologist. PMID:26045655

  8. Management of exfoliative glaucoma: challenges and solutions.

    PubMed

    Holló, Gábor; Katsanos, Andreas; Konstas, Anastasios Gp

    2015-01-01

    Exfoliative glaucoma is the most common type of secondary open-angle glaucoma worldwide. It is characterized by high intraocular pressure (IOP) and worse 24-hour IOP characteristics. In order to minimize progression, treatment of exfoliative glaucoma has to provide a low long-term mean IOP and good 24-hour IOP control. To achieve these goals, fixed-dose combination eye drops, argon and selective laser trabeculoplasty, and various forms of surgery (trabeculectomy, deep sclerectomy, viscocanalostomy, ab interno trabeculotomy, trabecular aspiration, and cataract surgery) all need to be considered during the long-term management of the disease. Since exfoliative glaucoma is a disease of the elderly, and is frequently associated with systemic vascular disease, interdisciplinary consultations are of great clinical importance. These management aspects and the current medical, laser, and surgical results are covered in this review, with a special focus on the needs of the general ophthalmologist.

  9. A Conserved Structural Motif Mediates Retrograde Trafficking of Shiga Toxin Types 1 and 2.

    PubMed

    Selyunin, Andrey S; Mukhopadhyay, Somshuvra

    2015-12-01

    Shiga toxin-producing Escherichia coli (STEC) produce two types of Shiga toxin (STx): STx1 and STx2. The toxin A-subunits block protein synthesis, while the B-subunits mediate retrograde trafficking. STEC infections do not have definitive treatments, and there is growing interest in generating toxin transport inhibitors for therapy. However, a comprehensive understanding of the mechanisms of toxin trafficking is essential for drug development. While STx2 is more toxic in vivo, prior studies focused on STx1 B-subunit (STx1B) trafficking. Here, we show that, compared with STx1B, trafficking of the B-subunit of STx2 (STx2B) to the Golgi occurs with slower kinetics. Despite this difference, similar to STx1B, endosome-to-Golgi transport of STx2B does not involve transit through degradative late endosomes and is dependent on dynamin II, epsinR, retromer and syntaxin5. Importantly, additional experiments show that a surface-exposed loop in STx2B (β4-β5 loop) is required for its endosome-to-Golgi trafficking. We previously demonstrated that residues in the corresponding β4-β5 loop of STx1B are required for interaction with GPP130, the STx1B-specific endosomal receptor, and for endosome-to-Golgi transport. Overall, STx1B and STx2B share a common pathway and use a similar structural motif to traffic to the Golgi, suggesting that the underlying mechanisms of endosomal sorting may be evolutionarily conserved.

  10. The Role of Botulinum Toxin Type A in the Clinical Management of Refractory Anterior Knee Pain

    PubMed Central

    Singer, Barbara J.; Silbert, Benjamin I.; Silbert, Peter L.; Singer, Kevin P.

    2015-01-01

    Anterior knee pain is a highly prevalent condition affecting largely young to middle aged adults. Symptoms can recur in more than two thirds of cases, often resulting in activity limitation and reduced participation in employment and recreational pursuits. Persistent anterior knee pain is difficult to treat and many individuals eventually consider a surgical intervention. Evidence for long term benefit of most conservative treatments or surgical approaches is currently lacking. Injection of Botulinum toxin type A to the distal region of vastus lateralis muscle causes a short term functional “denervation” which moderates the influence of vastus lateralis muscle on the knee extensor mechanism and increases the relative contribution of the vastus medialis muscle. Initial data suggest that, compared with other interventions for anterior knee pain, Botulinum toxin type A injection, in combination with an active exercise programme, can lead to sustained relief of symptoms, reduced health care utilisation and increased activity participation. The procedure is less invasive than surgical intervention, relatively easy to perform, and is time- and cost-effective. Further studies, including larger randomized placebo-controlled trials, are required to confirm the effectiveness of Botulinum toxin type A injection for anterior knee pain and to elaborate the possible mechanisms underpinning pain and symptom relief. PMID:26308056

  11. The Role of Botulinum Toxin Type A in the Clinical Management of Refractory Anterior Knee Pain.

    PubMed

    Singer, Barbara J; Silbert, Benjamin I; Silbert, Peter L; Singer, Kevin P

    2015-08-25

    Anterior knee pain is a highly prevalent condition affecting largely young to middle aged adults. Symptoms can recur in more than two thirds of cases, often resulting in activity limitation and reduced participation in employment and recreational pursuits. Persistent anterior knee pain is difficult to treat and many individuals eventually consider a surgical intervention. Evidence for long term benefit of most conservative treatments or surgical approaches is currently lacking. Injection of Botulinum toxin type A to the distal region of vastus lateralis muscle causes a short term functional "denervation" which moderates the influence of vastus lateralis muscle on the knee extensor mechanism and increases the relative contribution of the vastus medialis muscle. Initial data suggest that, compared with other interventions for anterior knee pain, Botulinum toxin type A injection, in combination with an active exercise programme, can lead to sustained relief of symptoms, reduced health care utilisation and increased activity participation. The procedure is less invasive than surgical intervention, relatively easy to perform, and is time- and cost-effective. Further studies, including larger randomized placebo-controlled trials, are required to confirm the effectiveness of Botulinum toxin type A injection for anterior knee pain and to elaborate the possible mechanisms underpinning pain and symptom relief.

  12. New diphtheria toxin repressor types depicted in a Romanian collection of Corynebacterium diphtheriae isolates.

    PubMed

    Dinu, Sorin; Damian, Maria; Badell, Edgar; Dragomirescu, Cristiana Cerasella; Guiso, Nicole

    2014-10-01

    Corynebacterium diphtheriae is the etiological agent of diphtheria, a potential fatal disease caused by a corynephage toxin. The expression of this diphtheria toxin is controlled via an iron-dependent repressor with various functions (DtxR). Some mutations in the dtxR gene are associated with diminished activity or even with total loss of DtxR function. We conducted a molecular study to characterize the dtxR alleles harbored by 34 isolates of C. diphtheriae recovered from Romanian patients between 1961 and 2007. Three of the seven alleles identified in this study have not previously been described. Two new DtxR types were identified, one of which has an unusual polypeptide length. All the new DtxR types were found in toxigenic isolates, suggesting that they effectively regulate the expression of diphtheria toxin. Furthermore, one of the new DtxR identified was also found in a non-toxigenic isolate, making it a potential source of toxigenic isolates after lysogenic conversion. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Comparison between intramuscular and perimuscular injections of botulinum toxin type A.

    PubMed

    Campos, José H; Oliveira, Lise B; Queiroz, Taise O; Santos, Kleber P; Freitas, Francesca M

    2006-01-01

    Botulinum toxin type A (BTX-A) must be injected in the intramuscular area to exert its paralytic effect. The durability of the BTX-A effect varies in different patients, and this fact can result from different locations of the drug injection, for example, the muscle peripheral area (perimuscular). This study aimed to evaluate whether a difference exists in the effect duration of the muscle paralysis between intramuscular and perimuscular injections of BTX-A. This study used 18 male New Zealand rabbits divided into two groups (A and B) based on the location of the BTX-A injection. The group A animals received 10 units of BTX-A diluted with 0.1 ml of normal saline injected perimuscularly. The group B animals received the same dosage injected in the intramuscular area of the left masseter muscle. An electroneurophysiologic study was performed 1 week before the experiment for all the animals, then repeated 1, 4, and 8 weeks after the toxin injection. The amplitude values recorded in the masseter muscle were significantly lower in both groups throughout the study than the physiologic amplitude. The comparison between groups A and B did not show any statistically significant amplitude variations throughout the 8 weeks. No significant difference in the neuromuscular blockade induced by botulinum toxin type A was observed between injections into the muscle peripheral area and intramuscular injections.

  14. Botulinum toxin type A in motor nervous system: unexplained observations and new challenges.

    PubMed

    Matak, I; Lacković, Z; Relja, M

    2016-12-01

    In the motor system, botulinum toxin type A (BoNT/A) actions were classically attributed to its well-known peripheral anticholinergic actions in neuromuscular junctions. However, the enzymatic activity of BoNT/A, assessed by the detection of cleaved synaptosomal-associated protein 25 (SNAP-25), was recently detected in motor and sensory regions of the brainstem and spinal cord after toxin peripheral injection in rodents. In sensory regions, the function of BoNT/A activity is associated with its antinociceptive effects, while in motor regions we only know that BoNT/A activity is present. Is it possible that BoNT/A presence in central motor nuclei is without any function? In this brief review, we analyze this question. Limited data available in the literature warrant further investigations of BoNT/A actions in motor nervous system.

  15. Evaluation of different fluids for detection of Clostridium perfringens type D epsilon toxin in sheep with experimental enterotoxemia.

    PubMed

    Layana, Jorge E; Fernandez Miyakawa, Mariano E; Uzal, Francisco A

    2006-08-01

    Enterotoxemia caused by Clostridium perfringens type D is a highly lethal disease of sheep, goats and other ruminants. The diagnosis of this condition is usually confirmed by detection of epsilon toxin, a major exotoxin produced by C. perfringens types B and D, in the intestinal content of affected animals. It has been suggested that other body fluids can also be used for detection of epsilon toxin. This study was performed to evaluate the usefulness of intestinal content versus other body fluids in detecting epsilon toxin in cases of sheep enterotoxemia. Samples of duodenal, ileal and colon contents, pericardial and abdominal fluids, aqueous humor and urine from 15 sheep with experimentally induced enterotoxemia, were analysed for epsilon toxin using a capture ELISA. Epsilon toxin was detected in 92% of the samples of ileal content, 64% of the samples of duodenal content, 57% of the samples of colon content and in 7% of the samples of pericardial fluid and aqueous humor. No epsilon toxin was found in samples of abdominal fluid or urine from the animals with enterotoxemia or in any samples from six clinically healthy sheep used as negative controls. The results of this study indicate that with the diagnostic capture ELISA used, intestinal content (preferably ileum) should be used for C. perfringens type D epsilon toxin detection in suspected cases of sheep enterotoxemia.

  16. Muscle structure and stiffness assessment after botulinum toxin type A injection. A systematic review.

    PubMed

    Mathevon, L; Michel, F; Decavel, P; Fernandez, B; Parratte, B; Calmels, P

    2015-12-01

    Botulinum toxin type A manages spasticity disorders in neurological central diseases. Some studies have reported that it might induce muscle changes. We present a literature review abiding by the PRISMA statement guidelines. The purpose was to explore the structural and passive biomechanical muscle properties after botulinum toxin type A injections in healthy and spastic limb muscles, on animals and humans, as well as methods for evaluating these properties. We searched the PubMed and Cochrane Library databases using the following keywords: "Botulinum toxin" AND ("muscle structure" OR "muscle atrophy") and, "Botulinum toxin" AND "muscle elasticity". From the 228 initially identified articles, 21 articles were included. Histological analyses were performed, especially on animals. A neurogenic atrophy systematically occurred. In humans, one year after a single injection, the histological recovery remained incomplete. Furthermore, 2D ultrasound analyses showed a reduction of the gastrocnemius thickness and pennation angle. MRI volumetric analysis evidenced muscular atrophy six months or one year after a single injection. Passive muscle stiffness depends on these structural changes. On the short term, the biomechanical analysis showed an elastic modulus increase in animals whereas no change was recorded in humans. On the short term, ultrasound elastography imaging showed a decreased elastic modulus. To date, few data are available, but all show a structural and mechanical muscle impact post injections, specifically muscle atrophy which can linger over time. Further studies are necessary to validate this element, and the possibility of change must be taken into account particularly with repeated injections. Thus, in clinical practice, 2D ultrasound and ultrasound elastography are two non-invasive techniques that will help physicians to develop an efficient long term monitoring. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  17. Factors affecting growth and toxin production by Clostridium botulinum type E on irradiated (0. 3 Mrad) chicken skins

    SciTech Connect

    Firstenberg-Eden, R.; Rowley, D.B.; Shattuck, G.E.

    1982-05-01

    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 30/sup 0/C in the absence of characteristic spoilage. Unstressed spores germinated, multiplied, and produced toxin on vacuum-packed chicken skins, stored at either 30 or 10/sup 0/C. Cell numbers increased faster and toxin was evident sooner at 30/sup 0/C than at 10/sup 0/C. At 30/sup 0/C, growth occurred and toxin was produced more slowly when samples were incubated aerobically than anaerobically. When samples were incubated aerobically at 10/sup 0/C, no toxin was detected within a test period of 14 days. An irradiation dose of 0.3 Mrad at 5/sup 0/C reduced a spore population on vacuum-sealed chicken skins by about 90%. The surviving population produced toxin at 30/sup 0/C under either aerobic or anaerobic conditions, at 10/sup 0/C no toxin was detected even on skins incubated anaerobically. Under the worst conditions (30/sup 0/C, vacuum packed) toxin was not detected prior to characteristic spoilage caused by the natural flora surviving 0.3 Mrad.

  18. Efficacy of type a botulinum toxin injections and infrared polarized light on treating chronic migraine.

    PubMed

    Song, J-H; Zhang, G-B; Ding, X-D; Huang, L; Hong, Y; Chen, H-X

    2015-01-01

    To investigate the clinical value of the combination of ultrasound-and-hyponome-guided type A botulinum toxin injection and infrared polarized light on treating chronic migraine. Ninety-one patients with chronic migraine were randomly divided into four groups: in the control group (group A, 22 cases in total), nimodipine was used in the treatment of chronic migraine for two months; in the infrared polarized light therapy group (group B, 22 cases in total), infrared polarized light was adopted in the treatment of chronic migraine for 50-60d; in the botulinum toxin treatment group (group C, 24 cases in total), ultrasound-and-hyponome-guided type A botulinum toxin was injected into frontal, temporal, and occipital muscles in treating chronic migraine; in the joint treatment group (group D, 23 cases in total), ultrasound-and-hyponome-guided type A botulinum toxin injection in group C and infrared polarized light in group B were both used here in the treatment of chronic migraine. Infrared polarized light therapy lasted 50-60d and the time of study lasted six months. The survey would include the conditions of patients with chronic migraine three months before treatment and at one, three and six months after treatment. Patients were asked to fill the MIDAS (migraine disability assessment questionnaire) and were graded on the evaluation scale of life quality, so that the researchers would be able to compare attack frequency, duration of attack, attack severity, the use of painkillers and their recovery from chronic migraine, and then observe their adverse reactions. Eleven cases dropped out during the treatment, three cases in A group, two cases in group B, four cases in group C and two cases in group D. One, three and six months after treatment, the MIDAS scores in group A, B, C and D were significantly lower than before the treatment. Hence, the differences were statistically significant (p < 0.01). The scores in quality of life rating scale were significantly higher

  19. A Structure-Function Analysis of Shiga-Like Toxin Type 2 of Enterohemorrhagic Escherichia Coli

    DTIC Science & Technology

    1990-05-07

    the Shiga toxin of Shigella dysenteriae I and the Shiga-like toxins of EHEC and in the identification of the toxin receptors, the biophysical ...10 Immunoreactivity 13C4 m m N^ N^ 16E6 m NA m NA with 1 19G8 NA NA NA NA VlAb :̂ BC5 + + + - + + + + Shiga toxin Shiga

  20. Pseudomonas aeruginosa Type III Secretory Toxin ExoU and Its Predicted Homologs

    PubMed Central

    Sawa, Teiji; Hamaoka, Saeko; Kinoshita, Mao; Kainuma, Atsushi; Naito, Yoshifumi; Akiyama, Koichi; Kato, Hideya

    2016-01-01

    Pseudomonas aeruginosa ExoU, a type III secretory toxin and major virulence factor with patatin-like phospholipase activity, is responsible for acute lung injury and sepsis in immunocompromised patients. Through use of a recently updated bacterial genome database, protein sequences predicted to be homologous to Ps. aeruginosa ExoU were identified in 17 other Pseudomonas species (Ps. fluorescens, Ps. lundensis, Ps. weihenstephanensis, Ps. marginalis, Ps. rhodesiae, Ps. synxantha, Ps. libanensis, Ps. extremaustralis, Ps. veronii, Ps. simiae, Ps. trivialis, Ps. tolaasii, Ps. orientalis, Ps. taetrolens, Ps. syringae, Ps. viridiflava, and Ps. cannabina) and 8 Gram-negative bacteria from three other genera (Photorhabdus, Aeromonas, and Paludibacterium). In the alignment of the predicted primary amino acid sequences used for the phylogenetic analyses, both highly conserved and nonconserved parts of the toxin were discovered among the various species. Further comparative studies of the predicted ExoU homologs should provide us with more detailed information about the unique characteristics of the Ps. aeruginosa ExoU toxin. PMID:27792159

  1. Pseudomonas aeruginosa Type III Secretory Toxin ExoU and Its Predicted Homologs.

    PubMed

    Sawa, Teiji; Hamaoka, Saeko; Kinoshita, Mao; Kainuma, Atsushi; Naito, Yoshifumi; Akiyama, Koichi; Kato, Hideya

    2016-10-26

    Pseudomonas aeruginosa ExoU, a type III secretory toxin and major virulence factor with patatin-like phospholipase activity, is responsible for acute lung injury and sepsis in immunocompromised patients. Through use of a recently updated bacterial genome database, protein sequences predicted to be homologous to Ps. aeruginosa ExoU were identified in 17 other Pseudomonas species (Ps. fluorescens, Ps. lundensis, Ps. weihenstephanensis, Ps. marginalis, Ps. rhodesiae, Ps. synxantha, Ps. libanensis, Ps. extremaustralis, Ps. veronii, Ps. simiae, Ps. trivialis, Ps. tolaasii, Ps. orientalis, Ps. taetrolens, Ps. syringae, Ps. viridiflava, and Ps. cannabina) and 8 Gram-negative bacteria from three other genera (Photorhabdus, Aeromonas, and Paludibacterium). In the alignment of the predicted primary amino acid sequences used for the phylogenetic analyses, both highly conserved and nonconserved parts of the toxin were discovered among the various species. Further comparative studies of the predicted ExoU homologs should provide us with more detailed information about the unique characteristics of the Ps. aeruginosa ExoU toxin.

  2. Type II Toxin-Antitoxin Distribution and Adaptive Aspects on Xanthomonas Genomes: Focus on Xanthomonas citri

    PubMed Central

    Martins, Paula M. M.; Machado, Marcos A.; Silva, Nicholas V.; Takita, Marco A.; de Souza, Alessandra A.

    2016-01-01

    Prokaryotic toxin-antitoxin (TA) systems were first described as being designed to prevent plasmid loss in bacteria. However, with the increase in prokaryotic genome sequencing, recently many TAs have been found in bacterial chromosomes, having other biological functions, such as environmental stress response. To date, only few studies have focused on TA systems in phytopathogens, and their possible impact on the bacterial fitness. This may be especially important for pathogens like Xanthomonas spp., which live epiphytically before entering the host. In this study, we looked for TA systems in the genomes of 10 Xanthomonas strains. We verified that citrus-infecting pathovars have, on average, 50% more TAs than other Xanthomonas spp. and no genome harbors classical toxins such as MqsR, RelB, and HicA. Only one TA system (PIN_VapC-FitB-like/SpoVT_AbrB) was conserved among the Xanthomonas genomes, suggesting adaptive aspects concerning its broad occurrence. We also detected a trend of toxin gene loss in this genus, while the antitoxin gene was preferably maintained. This study discovers the quantitative and qualitative differences among the type II TA systems present in Xanthomonas spp., especially concerning the citrus-infecting strains. In addition, the antitoxin retention in the genomes is possibly related with the resistance mechanism of further TA infections as an anti-addiction system or might also be involved in regulation of certain specific genes. PMID:27242687

  3. Molecular cloning and expression of epsilon toxin from Clostridium perfringens type D and tests of animal immunization.

    PubMed

    Souza, A M; Reis, J K P; Assis, R A; Horta, C C; Siqueira, F F; Facchin, S; Alvarenga, E R; Castro, C S; Salvarani, F M; Silva, R O S; Pires, P S; Contigli, C; Lobato, F C F; Kalapothakis, E

    2010-02-18

    Epsilon toxin produced by Clostridium perfringens types B and D causes enterotoxemia in sheep, goats and calves. Enterotoxemia can cause acute or superacute disease, with sudden death of the affected animal. It provokes huge economic losses when large numbers of livestock are affected. Therapeutic intervention is challenging, because the disease progresses very rapidly. However, it can be prevented by immunization with specific immunogenic vaccines. We cloned the etx gene, encoding epsilon toxin, into vector pET-11a; recombinant epsilon toxin (rec-epsilon) was expressed in inclusion bodies and was used for animal immunization. Serum protection was evaluated and cross-serum neutralization tests were used to characterize the recombinant toxin. To analyze the potency of the toxin (as an antigen), rabbits were immunized with 50, 100 or 200 microg recombinant toxin, using aluminum hydroxide gel as an adjuvant. Titers of 10, 30 and 40 IU/mL were obtained, respectively. These titers were higher than the minimum level required by the European Pharmacopoeia (5 IU/mL) and by the USA Code of Federal Regulation (2 IU/mL). This rec-epsilon is a good candidate for vaccine production against enterotoxemia caused by epsilon toxin of C. perfringens type D.

  4. The chromosomal nature of LT-II enterotoxins solved: a lambdoid prophage encodes both LT-II and one of two novel pertussis-toxin-like toxin family members in type II enterotoxigenic Escherichia coli

    PubMed Central

    Jobling, Michael G.

    2016-01-01

    Heat-labile enterotoxins (LT) of enterotoxigenic Escherichia coli (ETEC) are structurally and functionally related to cholera toxin (CT). LT-I toxins are plasmid-encoded and flanked by IS elements, while LT-II toxins of type II ETEC are chromosomally encoded with flanking genes that appear phage related. Here, I determined the complete genomic sequence of the locus for the LT-IIa type strain SA53, and show that the LT-IIa genes are encoded by a 51 239 bp lambdoid prophage integrated at the rac locus, the site of a defective prophage in E. coli K12 strains. Of 50 LT-IIa and LT-IIc, 46 prophages also encode one member of two novel two-gene ADP-ribosyltransferase toxin families that are both related to pertussis toxin, which I named eplBA or ealAB, respectively. The eplBA and ealAB genes are syntenic with the Shiga toxin loci in their lambdoid prophages of the enteric pathogen enterohemorrhagic E. coli. These novel AB5 toxins show pertussis-toxin-like activity on tissue culture cells, and like pertussis toxin bind to sialic acid containing glycoprotein ligands. Type II ETEC are the first mucosal pathogens known to simultaneously produce two ADP-ribosylating toxins predicted to act on and modulate activity of both stimulatory and inhibitory alpha subunits of host cell heterotrimeric G-proteins. PMID:26755534

  5. The chromosomal nature of LT-II enterotoxins solved: a lambdoid prophage encodes both LT-II and one of two novel pertussis-toxin-like toxin family members in type II enterotoxigenic Escherichia coli.

    PubMed

    Jobling, Michael G

    2016-04-01

    Heat-labile enterotoxins (LT) of enterotoxigenic Escherichia coli (ETEC) are structurally and functionally related to cholera toxin (CT). LT-I toxins are plasmid-encoded and flanked by IS elements, while LT-II toxins of type II ETEC are chromosomally encoded with flanking genes that appear phage related. Here, I determined the complete genomic sequence of the locus for the LT-IIa type strain SA53, and show that the LT-IIa genes are encoded by a 51 239 bp lambdoid prophage integrated at the rac locus, the site of a defective prophage in E. coli K12 strains. Of 50 LT-IIa and LT-IIc, 46 prophages also encode one member of two novel two-gene ADP-ribosyltransferase toxin families that are both related to pertussis toxin, which I named eplBA or ealAB, respectively. The eplBA and ealAB genes are syntenic with the Shiga toxin loci in their lambdoid prophages of the enteric pathogen enterohemorrhagic E. coli. These novel AB(5) toxins show pertussis-toxin-like activity on tissue culture cells, and like pertussis toxin bind to sialic acid containing glycoprotein ligands. Type II ETEC are the first mucosal pathogens known to simultaneously produce two ADP-ribosylating toxins predicted to act on and modulate activity of both stimulatory and inhibitory alpha subunits of host cell heterotrimeric G-proteins.

  6. Effect of botulinum toxin type A on gait of children who are idiopathic toe-walkers.

    PubMed

    Brunt, Denis; Woo, Raymund; Kim, Hyeong Dong; Ko, Man Soo; Senesac, Claudia; Li, Shuman

    2004-01-01

    The purpose of this study was to determine the effects of botulinum toxin type A treatment on ankle muscle activity during gait of children who are idiopathic toe-walkers. Five children who were idiopathic toe-walkers with a mean age was 4.34 years participated. Gait of the subjects was evaluated prior to, 20 days following, and 12 months following bilateral botulinum toxin type A injection of the gastrocnemius and soleus muscles. Subjects received physical therapy following the 20-day evaluation. Dependent variables were type of foot contact pattern and duration of swing-phase tibialis anterior activity and onset of stance-phase gastrocnemius relative to ground contact. Prior to treatment 51% of foot contacts were with the toe (heel just off the ground) or were digitigrade, while the remaining contacts were flat foot or heel strike. At approximately 20 days following treatment, only 8% of foot contacts were toe contact or digitigrade. Prior to treatment, mean gastrocnemius onset was 30 ms prior to foot contact and the duration of swing-phase tibialis anterior was only 345 ms. Following treatment (and a more normal foot contact pattern), mean gastrocnemius onset followed ground contact by 36 ms and tibialis anterior duration increased through terminal swing and into the loading response. The posttreatment improvement was maintained at 12-month follow-up. It appears that botulinum toxin type A treatment normalizes the ankle EMG pattern during gait and a more normal foot-strike pattern is obtained. These data are discussed in terms of a neuromotor rationale for the rehabilitation of children who are idiopathic toe-walkers to maintain posttreatment improvements.

  7. Is botulinum toxin type a intraprostatic injections really effective in patients with urinary retention?

    PubMed

    Jasiński, Miłosz; Drewa, Tomasz; Tyloch, Janusz; Wolski, Zbigniew

    2011-01-01

    Botulinum toxin type A is used in treatment of bladder hyperactivity and sphincter dyssynergia and was reported to alleviate lower urinary tract symptoms in patients with BPH. Some authors, however, failed to observe in their study apoptosis after BoNTA administration. We conducted an open-label study of BoNTA in men with BPH-related LUTS who were unsuitable for surgery as well as investigation of the effect of the toxin on in vitro growth of fibroblasts. In the clinical part, 5 patients aged from 75 to 88, suffering from BPH and UR were treated. Patients were previously disqualified from surgery and had not passed trials without catheters (TWOC). Prostate volume ranged from 38 to 104 mL. Botulinum toxin injection were performed. Each lobe of adenoma was injected with 100 U Botox under sonographic guidance. Prostate volume and TWOC were performed after 6 months. In the in vitro part, 3T3 mouse fibroblasts and fibroblasts isolated from human prostate were cultured in the presence of Botox (10, 5 and 1 U/mL) for 24 and 72 h. Cells were detached and counted in Neubauer chamber using trypan blue assay. Cells cultured in medium without botulinum toxin were the control group. Results are presented as the means with standard deviations. The means were compared, p <0.05 was considered statistically significant.No early complications were observed. Prostate volume remained unchanged after six months and patients were unable to void. Number of 3T3 cells after 24 h incubation was 7.12 +/- 1.88, 7.12 +/- 0.64, 6.75 +/- 1.28 and 6.88 +/- 0.83 x 10(4), after 24 h, 24.00 +/- 3.46, 22.75 +/- 3.73, 23.12 +/- 3.46 and 23.88 +/- 2.42 x 10(4) after 72 h, for 0, 1, 5 and 10 U/mL botulinum toxin type A concentrations, respectively. Similarly, number of prostate fibroblasts was 7.50 +/- 1.20, 7.12 +/- 1.73, 6.50 +/-1.93, and 6.25 +/- 1.58 x 10(4) after 24 h and 9.62 +/- 2.00, 9.12 +/- 1.55, 9.12 +/- 1.73 and 9.75 +/- 2.82 x 10(4) after 72 h. In conclusion, Botox had no statistically

  8. A comparison of outcome of surgical treatment of migraine headaches using a constellation of symptoms versus botulinum toxin type A to identify the trigger sites.

    PubMed

    Liu, Mengyuan T; Armijo, Bryan S; Guyuron, Bahman

    2012-02-01

    This study was designed to assess whether preoperative trigger-site confirmation using botulinum toxin type A injections significantly improved migraine surgery outcomes. The medical charts of 335 migraine surgery patients were reviewed. Patients who received stepwise diagnostic botulinum toxin type A injections were placed in the botulinum toxin type A group (n = 245). Patients who did not receive botulinum toxin type A or received only therapeutic botulinum toxin type A were placed in the control group (n = 90). The preoperative and 12-month postoperative migraine headache frequency, duration, and intensity were compared to determine the success of the operations. Seventy-two of 90 control patients (80 percent) experienced a significant improvement (a decrease of at least 50 percent in migraine headache frequency, duration, or intensity) at 12 months after surgery, with 29 (32 percent) reporting complete elimination. Of the 245 botulinum toxin type A patients, 207 (84 percent) experienced a significant improvement, with 89 (36 percent) experiencing complete elimination. The surgical success rate of the botulinum toxin type A group was not significantly higher than that of the control group (p = 0.33). Confirmation of trigger sites using botulinum toxin type A does not significantly improve the outcome of migraine surgery. Although botulinum toxin type A can be a useful diagnostic tool, this study demonstrates that there is no statistically significant difference between the injection of botulinum toxin type A and the use of a constellation of symptoms to identify trigger sites. Therapeutic, III.

  9. Muscle fiber-type changes induced by botulinum toxin injection in the rat larynx.

    PubMed

    Inagi, K; Connor, N P; Schultz, E; Ford, C N; Cook, C H; Heisey, D M

    1999-06-01

    This study examined muscle fiber-type alterations after single or multiple botulinum toxin (BT) injections to better understand possible morphologic changes induced by therapeutic BT injections in patients with spasmodic dysphonia. Muscle fiber staining was accomplished in rat intrinsic laryngeal muscles with antibodies to specific myosin heavy chains. Results indicated that the typical baseline distributions of type II muscle fibers (ie, types IIa, IIb, IIx, and IIL) were altered by BT injection, while no change was observed in type I fibers. Embryonic fibers were observed only along the needle insertion site at 7 days post BT injection. Although inferences from these animal data to human neuromuscular function must be made with caution, our findings provide insight into the possible cellular and molecular changes characterizing BT-injected muscles.

  10. Discovery of a Distinct Superfamily of Kunitz-Type Toxin (KTT) from Tarantulas

    PubMed Central

    Diao, Jian-Bo; Jiang, Li-Ping; Tang, Xing; Liang, Song-Ping

    2008-01-01

    Background Kuntiz-type toxins (KTTs) have been found in the venom of animals such as snake, cone snail and sea anemone. The main ancestral function of Kunitz-type proteins was the inhibition of a diverse array of serine proteases, while toxic activities (such as ion-channel blocking) were developed under a variety of Darwinian selection pressures. How new functions were grafted onto an old protein scaffold and what effect Darwinian selection pressures had on KTT evolution remains a puzzle. Principal Findings Here we report the presence of a new superfamily of KTTs in spiders (Tarantulas: Ornithoctonus huwena and Ornithoctonus hainana), which share low sequence similarity to known KTTs and is clustered in a distinct clade in the phylogenetic tree of KTT evolution. The representative molecule of spider KTTs, HWTX-XI, purified from the venom of O. huwena, is a bi-functional protein which is a very potent trypsin inhibitor (about 30-fold more strong than BPTI) as well as a weak Kv1.1 potassium channel blocker. Structural analysis of HWTX-XI in 3-D by NMR together with comparative function analysis of 18 expressed mutants of this toxin revealed two separate sites, corresponding to these two activities, located on the two ends of the cone-shape molecule of HWTX-XI. Comparison of non-synonymous/synonymous mutation ratios (ω) for each site in spider and snake KTTs, as well as PBTI like body Kunitz proteins revealed high Darwinian selection pressure on the binding sites for Kv channels and serine proteases in snake, while only on the proteases in spider and none detected in body proteins, suggesting different rates and patterns of evolution among them. The results also revealed a series of key events in the history of spider KTT evolution, including the formation of a novel KTT family (named sub-Kuntiz-type toxins) derived from the ancestral native KTTs with the loss of the second disulfide bridge accompanied by several dramatic sequence modifications. Conclusions

  11. Activity of a recombinant fusion protein between transforming growth factor type alpha and Pseudomonas toxin.

    PubMed Central

    Chaudhary, V K; FitzGerald, D J; Adhya, S; Pastan, I

    1987-01-01

    The transforming growth factor type alpha gene has been fused to a modified Pseudomonas toxin gene from which the cell-recognition domain has been deleted. The chimeric gene has been expressed in Escherichia coli, and the chimeric protein, PE40-TGF-alpha, has been highly purified. PE40-TGF-alpha kills cells expressing epidermal growth factor receptors and has little activity against cells with few receptors. This chimeric protein might be useful in treating cancers that contain high numbers of epidermal growth factor receptors. Images PMID:3299371

  12. Recurrent TMJ Dislocation Managed with Botulinum Toxin Type A Injections in a Pediatric Patient.

    PubMed

    Stark, Thomas R; Perez, Cristina V; Okeson, Jeffrey P

    2015-01-01

    Chronic recurrent temporomandibular joint (TMJ) dislocation is an uncommon condition that is painful and distressing to patients and uniquely challenging for clinicians. Sustained TMJ dislocation is not amenable to manual reduction alone when the etiology is muscular in nature. The purpose of this report was to describe the case of a child presenting with recurring temporomandibular joint dislocation secondary to muscle hyperactivity of unknown etiology that was managed with injections of botulinum toxin type A into the inferior lateral pterygoid muscles. The use of this peripheral antispasmoic neurotoxin is a reasonable, safe, and conservative, palliative treatment option for pediatric patients suffering from chronic recurring TMJ dislocation.

  13. The binding of cholera toxin to the periplasmic vestibule of the type II secretion channel

    PubMed Central

    Gonen, Melissa; Sun, Ji; Delarosa, Jaclyn R

    2011-01-01

    The type II secretion system (T2SS) is a large macromolecular complex spanning the inner and outer membranes of many Gram-negative bacteria. The T2SS is responsible for the secretion of virulence factors such as cholera toxin (CT) and heat-labile enterotoxin (LT) from Vibrio cholerae and enterotoxigenic Escherichia coli, respectively. CT and LT are closely related AB5 heterohexamers, composed of one A subunit and a B-pentamer. Both CT and LT are translocated, as folded protein complexes, from the periplasm across the outer membrane through the type II secretion channel, the secretin GspD. We recently published the 19 Å structure of the V. cholerae secretin (VcGspD) in its closed state and showed by SPR measurements that the periplasmic domain of GspD interacts with the B-pentamer complex. Here we extend these studies by characterizing the binding of the cholera toxin B-pentamer to VcGspD using electron microscopy of negatively stained preparations. Our studies indicate that the pentamer is captured within the large periplasmic vestibule of VcGspD. These new results agree well with our previously published studies and are in accord with a piston-driven type II secretion mechanism. PMID:21406971

  14. Monoclonal antibody-based immunoassay for type A Clostridium botulinum toxin is comparable to the mouse bioassay.

    PubMed Central

    Shone, C; Wilton-Smith, P; Appleton, N; Hambleton, P; Modi, N; Gatley, S; Melling, J

    1985-01-01

    A monoclonal antibody (BA11) has been produced against Clostridium botulinum type A neurotoxin by the fusion of myeloma cells (P3 NS1/1-Ag4-1) with spleen cells from BALB/c mice immunized with botulinum type A neurotoxoid. The antibody bound specifically to botulinum type A neurotoxin, showing no cross-reactivity with types B and E botulinum toxins or with any of several other bacterial toxins tested. The monoclonal antibody did not bind to botulinum type A neurotoxin which had been denatured with sodium dodecyl sulfate and bound only weakly to each of the separated heavy and light subunits of the neurotoxin, suggesting a conformational requirement for the antigenic determinant of the antibody. A sensitive immunoassay for C. botulinum type A toxin with monoclonal antibody BA11 in conjunction with an enzyme amplication system has been developed which allows detection of 5 to 10 mouse 50% lethal doses ml-1 of purified neurotoxin. The assay was equally sensitive when applied to the detection of crude toxin in food stuffs; the average value for the minimum level of detectable toxin in extracts of tinned salmon or corned beef was 9 +/- 3.1 mouse 50% lethal doses ml-1. PMID:3927840

  15. Rapid detection of vip1-type genes from Bacillus cereus and characterization of a novel vip binary toxin gene.

    PubMed

    Yu, Xiumei; Liu, Tao; Liang, Xiaoxing; Tang, Changqing; Zhu, Jun; Wang, Shiquan; Li, Shuangcheng; Deng, Qiming; Wang, Linxia; Zheng, Aiping; Li, Ping

    2011-12-01

    A PCR-restriction fragment length polymorphism (PCR-RFLP) method for identifying vegetative insecticidal protein (vip) 1-type genes from Bacillus cereus was developed by designing specific primers based on the conserved regions of the genes to amplify vip1-type gene fragments. PCR products were digested with endonuclease AciI, and four known vip1-type genes were identified. Vip1Ac and vip1Aa-type genes appeared in 17 of 26 B. cereus strains. A novel vip1-type gene, vip1Ac1, was identified from B. cereus strain HL12. The vip1Ac1 and vip2Ae3 genes were co-expressed in Escherichia coli strain BL21 by vector pCOLADuet-1. The binary toxin showed activity only against Aphis gossypii (Homoptera), but not for Coleptera (Tenebrio molitor, Holotrichia oblita), Lepidoptera (Spodoptera exigua, Helicoverpa armigera, and Chilo suppressalis), Diptera (Culex quinquefasciatus). The LC(50) of this binary toxin for A. gossypii is 87.5 (34.2-145.3) ng mL(-1) . This is probably only the second report that Vip1 and Vip2 binary toxin shows toxicity against homopteran pests. The PCR-RFLP method developed could be very useful for identifying novel Vip1-Vip2-type binary toxins, and the novel binary toxins, Vip1Ac1 and Vip2Ae3, identified in this study may have applications in biological control of insects, thus avoiding potential problems of resistance.

  16. Botulinum toxin A injection for spasticity in diplegic-type cerebral palsy.

    PubMed

    El, Ozlem; Peker, Ozlen; Kosay, Can; Iyilikci, Leyla; Bozan, Ozgur; Berk, Haluk

    2006-12-01

    Botulinum toxin type A can be both safe and effective in relieving spasticity in pediatric patients with cerebral palsy. In our prospective study, we evaluated the functional effect of botulinum toxin A in spastic diplegic-type cerebral palsy. Patients were examined on enrollment and at 1, 3, and 6 months after injection. Passive dorsiflexion of the ankle joint was measured using a goniometer as an angle of possible maximal dorsiflexion with the knee extended and flexed. Spasticity was graded using the Modified Ashworth Scale. Selective motor control at the ankle was assessed, and observational gait analysis was done. The functional status of the patients was determined by using the gross motor classification system. Botulinum toxin A was injected into the gastrocnemius muscle in all patients, and in four patients with concomitant jump knee gait, a hamstring muscle injection was added. Fourteen patients were included in the study. The mean age was 58.81 +/- 15.34 months. Following injection, spasticity was clinically decreased and statistically significant improvement was noticed in all clinical parameters after 1, 3, and 6 months of injection. The improvement in the clinical parameters decreased after 6 months but not to the baseline. One patient was Level II, four patients were Level III, and six patients were Level IV according to the Gross Motor Function Classification System at baseline. Improvement in the gross motor classification system is continued after 6 months in 12 children. The main goal of spasticity treatment in cerebral palsy is functional improvement. In our study, most of our patients had functional improvement according to the gross motor function classification system and did not change at 6 months.

  17. Hydrogen Implants for Layer Exfoliation

    NASA Astrophysics Data System (ADS)

    Cherekdjian, S.; Couillard, J. G.; Wilcox, C.

    2011-01-01

    Researchers at Corning Incorporated have developed a process whereby single crystal silicon thin films are transferred onto a flat panel display glass substrate using hydrogen ion implantation. The energy of the implant controls the effective exfoliation thickness, agreeing well with SRIM calculations, while the hydrogen ion dose controls the size of the platelets formed. The ion dose was found to influence the final void defect count in exfoliated films. Finally, the ion beam and ion implant end-station cooling characteristics were investigated. These parameters control the effective implant heat load generated during ion beam processing. The temperature at which exfoliation occurs during an exfoliation heat cycle was found to be inversely proportional to the hydrogen ion dose when the temperature during ion implantation is <100 °C. The most sensitive exfoliation temperature to ion dose dependence was observed for cooler implants, i.e. <35 °C. Data indicates that at the minimum exfoliation dose the exfoliation temperature is reduced significantly by increasing the implant heat generated during ion beam processing. Higher hydrogen doses than the minimum required for exfoliation exhibit only a small exfoliation temperature variation with ion dose. By optimizing the implant heat load generated during ion beam processing it is observed that the efficiency of the exfoliation process is also enhanced. Implant temperatures of 150 to 160 °C were found to further reduce the minimum implant dose required for exfoliation by an additional 5%, as verified by calorimetric measurements. These results enable us to further conclude that hydrogen out-diffusion is not significant in this process.

  18. Production of toxin by Clostridium botulinum type A strains cured by plasmids.

    PubMed Central

    Weickert, M J; Chambliss, G H; Sugiyama, H

    1986-01-01

    Twelve strains of Clostridium botulinum type A and seven strains of Clostridium sporogenes were screened for plasmids by agarose gel electrophoresis of cleared lysates of cells from 5 ml of mid-log-phase culture. Nine type A strains had one or more plasmids of 4.3, 6.8, or 36 megadaltons (MDa); several strains showed a large plasmid of 61 MDa, but it was not consistently recovered. Four C. sporogenes strains had one or more plasmids of 4.3, 5.6 or 36 MDa. Isolates obtained from cultures of plasmid-containing C. botulinum type A strains grown in ionic detergent broth and from spontaneously arising variants were screened both for toxin production and for plasmid content. Toxigenicity of C. botulinum could not be correlated with the presence of any one plasmid. Images PMID:3082278

  19. Glycosylphosphatidylinositols are potential targets for the development of novel inhibitors for aerolysin-type of pore-forming bacterial toxins.

    PubMed

    Wu, Qiuye; Guo, Zhongwu

    2010-03-01

    Many bacteria produce toxins that cause damage through the formation of pores in the host cell membrane. Some of these toxins, such as aerolysin, use glycosylphosphatidylinositols (GPIs) as their binding receptors to assist the pore formation on the host cell surface and the subsequent insertion of the resultant pores into the cell membrane. GPIs are a class of complex glycolipids that anchor surface proteins and glycoproteins onto the cell membrane in eukaryotic species. This review has summarized the reported evidences supporting the GPI-dependent pore-forming mechanism for aerolysin-type of toxins and analyzed the possibility of targeting this unique process for the design and development of novel GPI-based inhibitors for these pore-forming bacterial toxins. (c) 2009 Wiley Periodicals, Inc.

  20. Establishment of alternative potency test for botulinum toxin type A using compound muscle action potential (CMAP) in rats.

    PubMed

    Torii, Yasushi; Goto, Yoshitaka; Nakahira, Shinji; Ginnaga, Akihiro

    2014-11-01

    The biological activity of botulinum toxin type A has been evaluated using the mouse intraperitoneal (ip) LD50 test. This method requires a large number of mice to precisely determine toxin activity, and, as such, poses problems with regard to animal welfare. We previously developed a compound muscle action potential (CMAP) assay using rats as an alternative method to the mouse ip LD50 test. In this study, to evaluate this quantitative method of measuring toxin activity using CMAP, we assessed the parameters necessary for quantitative tests according to ICH Q2 (R1). This assay could be used to evaluate the activity of the toxin, even when inactive toxin was mixed with the sample. To reduce the number of animals needed, this assay was set to measure two samples per animal. Linearity was detected over a range of 0.1-12.8 U/mL, and the measurement range was set at 0.4-6.4 U/mL. The results for accuracy and precision showed low variability. The body weight was selected as a variable factor, but it showed no effect on the CMAP amplitude. In this study, potency tests using the rat CMAP assay of botulinum toxin type A demonstrated that it met the criteria for a quantitative analysis method.

  1. Chemoselective tarantula toxins report voltage activation of wild-type ion channels in live cells

    PubMed Central

    Tilley, Drew C.; Eum, Kenneth S.; Fletcher-Taylor, Sebastian; Austin, Daniel C.; Dupré, Christophe; Patrón, Lilian A.; Garcia, Rita L.; Lam, Kit; Yarov-Yarovoy, Vladimir; Cohen, Bruce E.; Sack, Jon T.

    2014-01-01

    Electrically excitable cells, such as neurons, exhibit tremendous diversity in their firing patterns, a consequence of the complex collection of ion channels present in any specific cell. Although numerous methods are capable of measuring cellular electrical signals, understanding which types of ion channels give rise to these signals remains a significant challenge. Here, we describe exogenous probes which use a novel mechanism to report activity of voltage-gated channels. We have synthesized chemoselective derivatives of the tarantula toxin guangxitoxin-1E (GxTX), an inhibitory cystine knot peptide that binds selectively to Kv2-type voltage gated potassium channels. We find that voltage activation of Kv2.1 channels triggers GxTX dissociation, and thus GxTX binding dynamically marks Kv2 activation. We identify GxTX residues that can be replaced by thiol- or alkyne-bearing amino acids, without disrupting toxin folding or activity, and chemoselectively ligate fluorophores or affinity probes to these sites. We find that GxTX–fluorophore conjugates colocalize with Kv2.1 clusters in live cells and are released from channels activated by voltage stimuli. Kv2.1 activation can be detected with concentrations of probe that have a trivial impact on cellular currents. Chemoselective GxTX mutants conjugated to dendrimeric beads likewise bind live cells expressing Kv2.1, and the beads are released by channel activation. These optical sensors of conformational change are prototype probes that can indicate when ion channels contribute to electrical signaling. PMID:25331865

  2. Chemoselective tarantula toxins report voltage activation of wild-type ion channels in live cells.

    PubMed

    Tilley, Drew C; Eum, Kenneth S; Fletcher-Taylor, Sebastian; Austin, Daniel C; Dupré, Christophe; Patrón, Lilian A; Garcia, Rita L; Lam, Kit; Yarov-Yarovoy, Vladimir; Cohen, Bruce E; Sack, Jon T

    2014-11-04

    Electrically excitable cells, such as neurons, exhibit tremendous diversity in their firing patterns, a consequence of the complex collection of ion channels present in any specific cell. Although numerous methods are capable of measuring cellular electrical signals, understanding which types of ion channels give rise to these signals remains a significant challenge. Here, we describe exogenous probes which use a novel mechanism to report activity of voltage-gated channels. We have synthesized chemoselective derivatives of the tarantula toxin guangxitoxin-1E (GxTX), an inhibitory cystine knot peptide that binds selectively to Kv2-type voltage gated potassium channels. We find that voltage activation of Kv2.1 channels triggers GxTX dissociation, and thus GxTX binding dynamically marks Kv2 activation. We identify GxTX residues that can be replaced by thiol- or alkyne-bearing amino acids, without disrupting toxin folding or activity, and chemoselectively ligate fluorophores or affinity probes to these sites. We find that GxTX-fluorophore conjugates colocalize with Kv2.1 clusters in live cells and are released from channels activated by voltage stimuli. Kv2.1 activation can be detected with concentrations of probe that have a trivial impact on cellular currents. Chemoselective GxTX mutants conjugated to dendrimeric beads likewise bind live cells expressing Kv2.1, and the beads are released by channel activation. These optical sensors of conformational change are prototype probes that can indicate when ion channels contribute to electrical signaling.

  3. Interaction of Type IV Toxin/Antitoxin Systems in Cryptic Prophages of Escherichia coli K-12

    PubMed Central

    Wen, Zhongling; Wang, Pengxia; Sun, Chenglong; Guo, Yunxue; Wang, Xiaoxue

    2017-01-01

    Toxin/antitoxin (TA) systems are widespread in prokaryotic chromosomes and in mobile genetic elements including plasmids and prophages. The first characterized Type IV TA system CbtA/CbeA was found in cryptic prophage CP4-44 in Escherichia coli K-12. Two homologous TA loci of CbtA/CbeA also reside in cryptic prophages of E. coli K-12, YkfI/YafW in CP4-6 and YpjF/YfjZ in CP4-57. In this study, we demonstrated that YkfI and YpjF inhibited cell growth and led to the formation of “lemon-shaped” cells. Prolonged overproduction of YkfI led to the formation of “gourd-shaped” cells and immediate cell lysis. YafW and YfjZ can neutralize the toxicity of YkfI or YpjF. Furthermore, we found that YkfI and YpjF interacted with cell division protein FtsZ in E. coli, but ectopic expression in Pseudomonas and Shewanella did not cause the formation of “lemon-shaped” cells. Moreover, deletion of all of the three toxin genes together decreased resistance to oxidative stress and deletion of the antitoxin genes increased early biofilm formation. Collectively, these results demonstrated that the homologous Type IV TA systems in E. coli may target cell division protein FtsZ in E. coli and may have different physiological functions in E. coli. PMID:28257056

  4. Botulinum toxin type A selectivity for certain types of pain is associated with capsaicin-sensitive neurons.

    PubMed

    Matak, Ivica; Rossetto, Ornella; Lacković, Zdravko

    2014-08-01

    Unlike most classical analgesics, botulinum toxin type A (BoNT/A) does not alter acute nociceptive thresholds, and shows selectivity primarily for allodynic and hyperalgesic responses in certain pain conditions. We hypothesized that this phenomenon might be explained by characterizing the sensory neurons targeted by BoNT/A in the central nervous system after its axonal transport. BoNT/A's central antinociceptive activity following its application into the rat whisker pad was examined in trigeminal nucleus caudalis (TNC) and higher-level nociceptive brain areas using BoNT/A-cleaved synaptosomal-associated protein 25 (SNAP-25) and c-Fos immunohistochemistry. Occurrence of cleaved SNAP-25 in TNC was examined after nonselective ganglion ablation with formalin or selective denervation of capsaicin-sensitive (vanilloid receptor-1 or TRPV1-expressing) neurons, and in relation to different cellular and neuronal markers. Regional c-Fos activation and effect of TRPV1-expressing afferent denervation on toxin's antinociceptive action were studied in formalin-induced orofacial pain. BoNT/A-cleaved SNAP-25 was observed in TNC, but not in higher-level nociceptive nuclei. Cleaved SNAP-25 in TNC disappeared after formalin-induced trigeminal ganglion ablation or capsaicin-induced sensory denervation. Occurrence of cleaved SNAP-25 in TNC and BoNT/A antinociceptive activity in formalin-induced orofacial pain were prevented by denervation with capsaicin. Cleaved SNAP-25 localization demonstrated toxin's presynaptic activity in TRPV1-expressing neurons. BoNT/A reduced the c-Fos activation in TNC, locus coeruleus, and periaqueductal gray. Present experiments suggest that BoNT/A alters the nociceptive transmission at the central synapse of primary afferents. Targeting of TRPV1-expressing neurons might be associated with observed selectivity of BoNT/A action only in certain types of pain.

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

    PubMed

    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. © 2010 European Paediatric Neurology Society. Published by Elsevier Ltd. All rights reserved.

  6. Development of a quail embryo model for the detection of botulinum toxin type A activity

    USDA-ARS?s Scientific Manuscript database

    Clostridium botulinum is a ubiquitous microorganism which under certain anaerobic conditions can produce botulinum toxins. Due to concerns in regards to both food-borne illness and the potential use of botulinum toxin as a biological weapon, the capability to assess the amount of toxin in a food or...

  7. Serial Casting as an Adjunct to Botulinum Toxin Type A Treatment in Children With Cerebral Palsy and Spastic Paraparesis With Scissoring of the Lower Extremities.

    PubMed

    Dai, Alper I; Demiryürek, Abdullah T

    2017-06-01

    The purpose of this study was to examine whether combination therapy of serial casting and botulinum toxin type A injection can further enhance the effects of botulinum toxin type A in children with cerebral palsy with scissoring of both legs. This study was a prospective and randomized trial. The children were divided into 2 groups, one of which received serial casting after botulinum toxin type A (n = 40), and the other which only received botulinum toxin type A (n = 40). Serial casting started 3 weeks after the botulinum toxin type A. Both groups received physiotherapy. Groups were assessed at baseline then compared at 6 and 12 weeks following the intervention. Significant improvements in Gross Motor Function Measure-66 and Caregiver Health Questionnaire were recorded in both groups ( P < .001). The modified Ashworth scale improved significantly following botulinum toxin type A in the serial casting group ( P < .05), but not in botulinum toxin type A only group. These results suggest that serial casting after botulinum toxin type A can enhance the benefits of botulinum toxin type A in children with cerebral palsy.

  8. Efficacy of DNA vaccines expressing the type F botulinum toxin Hc fragment using different promoters.

    PubMed

    Jathoul, Amit P; Holley, Jane L; Garmory, Helen S

    2004-09-28

    DNA vaccines which expressed the Hc fragment of the Clostridium botulinum type F neurotoxin (BoNT/F Hc) fused to a signal peptide downstream of four different eukaryotic promoters were prepared. Subsequently, the immunogenicity of the DNA vaccines and protection afforded in mice against challenge with 10(4) MLD of type F botulinum toxin was evaluated. The DNA vaccine containing the human ubiquitin gene (UbC) promoter induced the highest BoNT/F Hc-specific antibody concentration following two intramuscular immunisations and afforded 90% protection against challenge. The results from this study indicate that the selection of promoter used in DNA vaccination studies may be of importance in designing optimised vaccines.

  9. Efficacy of botulinum toxin in pachyonychia congenita type 1: report of two new cases.

    PubMed

    González-Ramos, Jéssica; Sendagorta-Cudós, Elena; González-López, Guillermo; Mayor-Ibarguren, Ander; Feltes-Ochoa, Rosa; Herranz-Pinto, Pedro

    2016-01-01

    Pachyonychia congenita (PC) is a rare genodermatosis caused by a mutation in keratin genes, which can lead to hypertrophic nail dystrophy and focal palmoplantar keratoderma (predominantly plantar), amongst other manifestations. Painful blisters and callosities, sometimes exacerbated by hyperhidrosis, are major issues that can have a significant impact on patient quality of life. Many alternative treatments for this condition have been applied with variable and partial clinical response, but a definitive cure for this disease has yet to be discovered. After obtaining informed consent, two patients with genetically confirmed PC type 1 were treated with plantar injections of botulinum toxin type A. Both patients showed a marked improvement in pain and blistering with an average response time of one week, a six-month mean duration of effectiveness, and a lack of any side effects or tachyphylaxis.

  10. Recompressed exfoliated graphite articles

    DOEpatents

    Zhamu, Aruna; Shi, Jinjun; Guo, Jiusheng; Jang, Bor Z

    2013-08-06

    This invention provides an electrically conductive, less anisotropic, recompressed exfoliated graphite article comprising a mixture of (a) expanded or exfoliated graphite flakes; and (b) particles of non-expandable graphite or carbon, wherein the non-expandable graphite or carbon particles are in the amount of between about 3% and about 70% by weight based on the total weight of the particles and the expanded graphite flakes combined; wherein the mixture is compressed to form the article having an apparent bulk density of from about 0.1 g/cm.sup.3 to about 2.0 g/cm.sup.3. The article exhibits a thickness-direction conductivity typically greater than 50 S/cm, more typically greater than 100 S/cm, and most typically greater than 200 S/cm. The article, when used in a thin foil or sheet form, can be a useful component in a sheet molding compound plate used as a fuel cell separator or flow field plate. The article may also be used as a current collector for a battery, supercapacitor, or any other electrochemical cell.

  11. Lack of anti-inflammatory effect of botulinum toxin type A in experimental models of inflammation.

    PubMed

    Bach-Rojecky, Lidija; Dominis, Mara; Lacković, Zdravko

    2008-10-01

    Botulinum toxin type A (BTX-A) has a long-lasting antinociceptive activity and less clear effect on inflammation. It was proposed that these two effects share the same mechanism--the inhibition of neurotransmitter exocytosis from peripheral nerve endings. However, till now possible anti-inflammatory action of BTX-A did not evoke much attention. In the present paper, we investigate possible anti-inflammatory action of the toxin in carrageenan and capsaicin models of inflammation in rats. BTX-A (5 and 10 U/kg) was injected into the plantar surface of the rat right hind-paw pad 5 days before the injection of the carrageenan (1%) or capsaicin (0.1%) at the same site. Carrageenan-induced paw oedema and capsaicin-induced protein extravasation were measured. Control, inflamed and BTX-A pretreated inflamed paws were photographed and histopathological analysis (haematoxylin & eosin) was performed. Pretreatment with BTX-A had no effect on the size of carrageenan-induced paw oedema, measured as paw volume and weight or capsaicin-induced plasma extravasations, measured by Evans blue as a marker of protein leakage. Neither macroscopic nor microscopic analysis showed a significant difference between BTX-A pretreated and control inflamed tissue. Results show dissociation between the effect of BTX-A on pain and inflammation thus questioning the validity of the suggested assumption about the common peripheral mechanism of action.

  12. Botulinum toxin type A for rehabilitation after a spinal cord injury: a case report.

    PubMed

    Naicker, A S; Roohi, S A; Chan, J L L

    2009-04-01

    A 56-year-old man became quadriplegic, bed bound, and carer-dependent secondary to cervical osteomyelitis. Three years later, he presented with generalised spasticity, crouched posture, and a large sacral pressure sore. The severe spasticity in his hips and knees prevented ischial sitting. Injections of botulinum toxin type A to both hamstrings and gastrosoleuii controlled the flexor spasticity of his lower limbs and facilitated rehabilitation and wound healing through proper positioning, wound care, stretching, and weight-bearing exercises. A few weeks later, the patient could better position himself in bed (prone lying) and on his wheelchair (ischial sitting). His spasm-related pain lessened and his mobility and activities of daily living improved. The sacral pressure sore healed completely a few months later. The patient could sleep better, feed with set-up and adaptive aids, groom, dress, and transfer himself with minimal assistance. The effects of botulinum toxin extended beyond just spasticity reduction. His upper extremity function, mobility, and social well-being were all improved through better positioning.

  13. Changes of masseter muscle activity following injection of botulinum toxin type A in adult rats.

    PubMed

    Tsai, C Y; Lei, Y Y; Yang, L Y; Chiu, W C

    2015-11-01

    To investigate changes in masseter muscle function following intramuscular injection of different dose-dependent botulinum toxin type A (BTXA). Department of Orthodontics at Taipei Medical University. Fifty-two, 70-day-old male Wistar rats were randomly divided into four groups. Group I received 7.5 U of BTXA (0.3 ml), Group II received 5.0 U, and Group III received 2.5 U in the right masseter muscle, respectively. Group IV is the control and received no BTXA injection. A wire electrode device was implanted to record muscle activity. One week after implantation, the rats were fed every 2 h and EMG signals were recorded during the first hour. All signals were recorded for 12 weeks. Thereafter, EMG data were analyzed for statistical calculation and weights of masseter muscles were measured. Masseter muscle activity decreased 99% during the first week after BTXA injection and gradually recovered from the 3rd week on in Groups I-III. By the 12th week, muscle activity recovered to 41% in Groups I and II and 56.26% in Group III. No significant changes of muscle activity were observed in Group IV. BTXA induced a reduction in masseter muscle activity and an increased toxin dose resulted in greater depression of muscle activity. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Epsilon Toxin Is Essential for the Virulence of Clostridium perfringens Type D Infection in Sheep, Goats, and Mice

    PubMed Central

    Garcia, J. P.; Adams, V.; Beingesser, J.; Hughes, M. L.; Poon, R.; Lyras, D.; Hill, A.; McClane, B. A.; Rood, J. I.

    2013-01-01

    Clostridium perfringens type D causes disease in sheep, goats, and other ruminants. Type D isolates produce, at minimum, alpha and epsilon (ETX) toxins, but some express up to five different toxins, raising questions about which toxins are necessary for the virulence of these bacteria. We evaluated the contribution of ETX to C. perfringens type D pathogenicity in an intraduodenal challenge model in sheep, goats, and mice using a virulent C. perfringens type D wild-type strain (WT), an isogenic ETX null mutant (etx mutant), and a strain where the etx mutation has been reversed (etx complemented). All sheep and goats, and most mice, challenged with the WT isolate developed acute clinical disease followed by death in most cases. Sheep developed various gross and/or histological changes that included edema of brain, lungs, and heart as well as hydropericardium. Goats developed various effects, including necrotizing colitis, pulmonary edema, and hydropericardium. No significant gross or histological abnormalities were observed in any mice infected with the WT strain. All sheep, goats, and mice challenged with the isogenic etx mutant remained clinically healthy for ≥24 h, and no gross or histological abnormalities were observed in those animals. Complementation of etx knockout restored virulence; most goats, sheep, and mice receiving this complemented mutant developed clinical and pathological changes similar to those observed in WT-infected animals. These results indicate that ETX is necessary for type D isolates to induce disease, supporting a key role for this toxin in type D disease pathogenesis. PMID:23630957

  15. CgNa, a type I toxin from the giant Caribbean sea anemone Condylactis gigantea shows structural similarities to both type I and II toxins, as well as distinctive structural and functional properties1

    PubMed Central

    Salceda, Emilio; Pérez-Castells, Javier; López-Méndez, Blanca; Garateix, Anoland; Salazar, Hector; López, Omar; Aneiros, Abel; Ständker, Ludger; Béress, Lászlo; Forssmann, Wolf-Georg; Soto, Enrique; Jiménez-Barbero, Jesús; Giménez-Gallego, Guillermo

    2007-01-01

    CgNa (Condylactis gigantea neurotoxin) is a 47-amino-acid- residue toxin from the giant Caribbean sea anemone Condylactis gigantea. The structure of CgNa, which was solved by 1H-NMR spectroscopy, is somewhat atypical and displays significant homology with both type I and II anemone toxins. CgNa also displays a considerable number of exceptions to the canonical structural elements that are thought to be essential for the activity of this group of toxins. Furthermore, unique residues in CgNa define a characteristic structure with strong negatively charged surface patches. These patches disrupt a surface-exposed cluster of hydrophobic residues present in all anemone-derived toxins described to date. A thorough characterization by patch–clamp analysis using rat DRG (dorsal root ganglion) neurons indicated that CgNa preferentially binds to TTX-S (tetrodotoxin-sensitive) voltage-gated sodium channels in the resting state. This association increased the inactivation time constant and the rate of recovery from inactivation, inducing a significant shift in the steady state of inactivation curve to the left. The specific structural features of CgNa may explain its weaker inhibitory capacity when compared with the other type I and II anemone toxins. PMID:17506725

  16. CgNa, a type I toxin from the giant Caribbean sea anemone Condylactis gigantea shows structural similarities to both type I and II toxins, as well as distinctive structural and functional properties(1).

    PubMed

    Salceda, Emilio; Pérez-Castells, Javier; López-Méndez, Blanca; Garateix, Anoland; Salazar, Hector; López, Omar; Aneiros, Abel; Ständker, Ludger; Béress, Lászlo; Forssmann, Wolf-Georg; Soto, Enrique; Jiménez-Barbero, Jesús; Giménez-Gallego, Guillermo

    2007-08-15

    CgNa (Condylactis gigantea neurotoxin) is a 47-amino-acid- residue toxin from the giant Caribbean sea anemone Condylactis gigantea. The structure of CgNa, which was solved by 1H-NMR spectroscopy, is somewhat atypical and displays significant homology with both type I and II anemone toxins. CgNa also displays a considerable number of exceptions to the canonical structural elements that are thought to be essential for the activity of this group of toxins. Furthermore, unique residues in CgNa define a characteristic structure with strong negatively charged surface patches. These patches disrupt a surface-exposed cluster of hydrophobic residues present in all anemone-derived toxins described to date. A thorough characterization by patch-clamp analysis using rat DRG (dorsal root ganglion) neurons indicated that CgNa preferentially binds to TTX-S (tetrodotoxin-sensitive) voltage-gated sodium channels in the resting state. This association increased the inactivation time constant and the rate of recovery from inactivation, inducing a significant shift in the steady state of inactivation curve to the left. The specific structural features of CgNa may explain its weaker inhibitory capacity when compared with the other type I and II anemone toxins.

  17. Molecular Resolution in situ Imaging of Spontaneous Graphene Exfoliation.

    PubMed

    Elbourne, Aaron; McLean, Ben; Voïtchovsky, Kislon; Warr, Gregory G; Atkin, Rob

    2016-08-18

    All reported methods of graphene exfoliation require external energy input, most commonly from sonication,1 shaking,2 or stirring.3 The reverse process-aggregation of single or few layer graphene sheets-occurs spontaneously in most solvents. This makes producing, and especially storing, graphene in economic quantities challenging,4,5 which is a significant barrier to widespread commercialization. This study reveals ionic liquids (ILs) can spontaneously exfoliate graphene from graphite at room temperature. The process is thermally activated and follows an Arrhenius-type behavior, resulting in thermodynamically stable IL/graphene suspensions. Using atomic force microscopy, the kinetics of the exfoliation could be followed in situ and with subnanometer resolution, showing that both the size and the charge of the constituent IL ions play a key role. Our results provide a general molecular mechanism underpinning spontaneous graphene exfoliation at room temperature in electrically conducting ILs, paving the way for their adoption in graphene-based technology.

  18. Targeting bacterial toxins.

    PubMed

    Ivarsson, Mattias E; Leroux, Jean-Christophe; Castagner, Bastien

    2012-04-23

    Protein toxins constitute the main virulence factors of several species of bacteria and have proven to be attractive targets for drug development. Lead candidates that target bacterial toxins range from small molecules to polymeric binders, and act at each of the multiple steps in the process of toxin-mediated pathogenicity. Despite recent and significant advances in the field, a rationally designed drug that targets toxins has yet to reach the market. This Review presents the state of the art in bacterial toxin targeted drug development with a critical consideration of achieved breakthroughs and withstanding challenges. The discussion focuses on A-B-type protein toxins secreted by four species of bacteria, namely Clostridium difficile (toxins A and B), Vibrio cholerae (cholera toxin), enterohemorrhagic Escherichia coli (Shiga toxin), and Bacillus anthracis (anthrax toxin), which are the causative agents of diseases for which treatments need to be improved. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. [The use of botulinum toxin type a in the acute phase of facial nerve injury after neurosurgical surgery].

    PubMed

    Orlova, O R; Akulov, M A; Usachev, D Iu; Taniashin, S V; Zakharov, V O; Saksonova, E V; Mingazova, L R; Surovykh, S V

    2014-01-01

    To evaluate the role of botulinum toxin type A in the acute phase of facial nerve injury after neurosurgical surgery. The study involved 55 patients with acute facial muscle paresis caused by facial nerve injury during surgery on the posterior cranial fossa and cerebello-pontine angle (CPA). The first group consisted of 35 patients (mean age, 48.14±1.26 years) who were administered botulinum toxin type A (xeomin) at a dose of 2-3 U per point in muscles of the intact side of the face. The control group included 20 patients (mean age, 49.85±1.4 years) who underwent standard rehabilitation treatment of this pathology. The treatment efficacy was evaluated using the House-Brackmann Scale, the Yanagihara facial grading system, the Facial Disability Index (FDI), and the Sunnybrook Facial Grading (SFG) Scale. Before treatment, patients of both groups experienced severe dysfunction according to the House-Brackmann Scale. A month after the botulinium toxin type A therapy had been started, a significant improvement in the group of patients who received botulinum toxin was observed at all scales (p<0.05), whereas improvement in the facial nerve function in the second group was observed only by the 3rd month of rehabilitation treatment (p<0.05). The number of synkineses in the patients who did not receive botulinum toxin was 46% higher than that in the first group (p=0.019) one year after the surgery, and it was higher by 91% after 2 years (p<0.001). The use of botulinum toxin type A is reasonable in acute facial nerve injury and should be mandatory in combined therapy of these patients.

  20. Critical analysis of the use of onabotulinumtoxinA (botulinum toxin type A) in migraine

    PubMed Central

    Robertson, Carrie E; Garza, Ivan

    2012-01-01

    OnabotulinumtoxinA, a neurotoxin, has been studied in numerous trials as a novel preventive therapy for migraine headache. The data would support that it may be effective at reducing headache days in patients suffering from chronic migraine (≥15 headache days/month, with eight or more of those migraine headache days). The mechanism by which onabotulinumtoxinA exerts its effects on migraine is not yet understood. It is known to inhibit acetylcholine release at the neuromuscular junction, but this probably does not explain the observed antinociceptive properties noted in preclinical and clinical trials. This review will discuss the known mechanisms of action of botulinum toxin type A, and will review the available randomized, placebo-controlled trials that have looked at its efficacy as a migraine preventative. We also describe the onabotulinumtoxinA injection sites used at our institution. PMID:22275844

  1. A plant-based oral vaccine to protect against systemic intoxication by Shiga toxin type 2.

    PubMed

    Wen, Sharon X; Teel, Louise D; Judge, Nicole A; O'Brien, Alison D

    2006-05-02

    Hemolytic uremic syndrome, the leading cause of kidney failure in children, often follows infection with enterohemorrhagic Escherichia coli and is mediated by the Shiga type toxins, particularly type 2 (Stx2), produced by such strains. The challenge in protecting against this life-threatening syndrome is to stimulate an immune response at the site of infection while also protecting against Shiga intoxication at distal sites such as the kidney. As one approach to meeting this challenge, we sought to develop and characterize a prototypic orally delivered, plant-based vaccine against Stx2, an AB5 toxin. First, we genetically inactivated the Stx2 active A subunit gene and then optimized both subunit genes for expression in plants. The toxoid genes were then transformed into the Nicotiana tabacum (tobacco) cell line NT-1 by Agrobacterium tumefaciens-mediated transformation. Toxoid expression was detected in NT-1 cell extracts, and the assembly of the holotoxoid was confirmed. Finally, mice were immunized by feeding with the toxoid-expressing NT-1 cells or by parenteral immunization followed by oral vaccination (prime-boost strategy). The immunized mice produced Stx2-specific mucosal IgA and Stx2-neutralizing serum IgG. The protective efficacy of these responses was assessed by challenging the immunized mice with E. coli O91:H21 strain B2F1, an isolate that produces an activatable variant of Stx2 (Stx2d) and is lethal to mice. The oral immunization fully protected mice from the challenge. Results of this study demonstrated that a plant-based oral vaccine can confer protection against lethal systemic intoxication.

  2. Genetic diversity of the mating type and toxin production genes in Pyrenophora tritici-repentis.

    PubMed

    Lepoint, P; Renard, M-E; Legrève, A; Duveiller, E; Maraite, H

    2010-05-01

    Pyrenophora tritici-repentis, the causal agent of tan spot on wheat, is a homothallic loculoascomycete with a complex race structure. The objectives of this study were to confirm the homothallic nature of the pathogen, characterize mating type diversity and toxin production genes in a global collection of strains, and analyze how these traits are associated between each other and with existing races. The pseudothecia production capacity, race identification, mating type locus (MAT), internal transcribed spacer, and glyceraldehyde-3-phosphate dehydrogenase regions were analyzed in a selection of 88 strains originating from Europe, North and South America, North Africa, and Central and South Asia. Some (60%) strains produced pseudothecia containing ascospores, independent of their origin. Race identification obtained using the multiplex polymerase chain reaction targeting host-selective toxin (HST) genes was consistent, overall, with the results based on the inoculation of a set of differential wheat cultivars and confirmed the predominance of race 1/2 strains ( approximately 83%). However, discrepancies in race identification, differences from the reference tester strains, and atypical ToxA profiles suggest the presence of new races and HSTs. The MAT1-1 and MAT1-2 coding regions are consecutively arranged in a single individual, suggesting putative heterothallic origin of P. tritici-repentis. Upstream from the MAT is an open reading frame of unknown function (ORF1) containing a MAT-specific degenerate carboxy-terminus. The phylogenetic analysis of the MAT locus reveals two distinct groups, unlinked to geographical origin or ToxA profile. Group I, the best-represented group, is associated with typical tan spot lesions caused by races 1, 2, 3, and 5 on wheat. It is more homogenous than group II encompassing race 4 strains, as well as isolates associated primarily with small spot lesions on wheat leaves or other hosts. Group II could contain several distinct taxa.

  3. Shiga Toxin (Stx) Type 1a Reduces the Oral Toxicity of Stx Type 2a

    PubMed Central

    Russo, Lisa M.; Melton-Celsa, Angela R.; O'Brien, Alison D.

    2016-01-01

    Background. Shiga toxin (Stx) is the primary virulence factor of Stx-producing Escherichia coli (STEC). STEC can produce Stx1a and/or Stx2a, which are antigenically distinct. However, Stx2a-producing STEC are associated with more severe disease than strains producing both Stx1a and Stx2a. Methods and Results. To address the hypothesis that the reason for the association of Stx2a with more severe disease is because Stx2a crosses the intestinal barrier with greater efficiency that Stx1a, we covalently labeled Stx1a and Stx2a with Alexa Fluor 750 and determined the ex vivo fluorescent intensity of murine systemic organs after oral intoxication. Surprisingly, both Stxs exhibited similar dissemination patterns and accumulated in the kidneys. We next cointoxicated mice to determine whether Stx1a could impede Stx2a. Cointoxication resulted in increased survival and an extended mean time to death, compared with intoxication with Stx2a only. The survival benefit was dose dependent, with the greatest effect observed when 5 times more Stx1a than Stx2a was delivered, and was amplified when Stx1a was delivered 3 hours prior to Stx2a. Cointoxication with an Stx1a active site toxoid also reduced Stx2a toxicity. Conclusions. These studies suggest that Stx1a reduces Stx2a-mediated toxicity, a finding that may explain why STEC that produce only Stx2a are associated with more severe disease than strains producing Stx1a and Stx2a. PMID:26743841

  4. [Detection of Clostridium novyi type B alpha toxin using cell culture systems

    PubMed

    Borrmann, Erika; Schulze, Frank

    1998-01-01

    The aim of our study was to investigate if a cell culture assay can replace the toxin neutralisation test in mice for the potency testing of alpha toxoid containing clostridial vaccines for veterinary use. The basis for the development of our cell culture assay was the detection of the cytotoxic/cytopathic action of alpha toxin on cells in culture. Nine permanent cell lines were examined for their reaction to the alpha toxin. The action of the toxin was determined after three days by microscopic examination and MTT assay. The alpha toxin exhibited the strongest effect on the ESH-L cells. We were able to show that the cytopathic effect was neutralised by the international standard for gas gangrene antitoxin (C.novyi) but never by heterologous antisera. Our results showed that the ESH-L cell line was a suitable indicator for the detection of the cytotoxic effect of alpha toxin.

  5. A novel toxin homologous to large clostridial cytotoxins found in culture supernatant of Clostridium perfringens type C.

    PubMed

    Amimoto, Katsuhiko; Noro, Taichi; Oishi, Eiji; Shimizu, Mitsugu

    2007-04-01

    An unknown cytotoxin was identified in the culture supernatant of Clostridium perfringens type C. The cytotoxin, named TpeL, which was purified using mAb-based affinity chromatography, had a lethal activity of 62 minimum lethal dose (MLD) mg(-1) in mice and a cytotoxic activity of 6.2x10(5) cytotoxic units (CU) mg(-1) in Vero cells. The nucleotide sequence of TpeL was determined. The entire ORF had a length of 4953 bases, and the same nucleotide sequence was not recorded in the GenBank/EMBL/DDBJ databases. The molecular mass calculated from the deduced amino acid sequence was 191 kDa, and a signal peptide region was not found within the ORF. The deduced amino acid sequence exhibited 30-39 % homology to Clostridium difficile toxins A (TcdA) and B (TcdB), Clostridium sordellii lethal toxin (TcsL) and Clostridium novyi alpha-toxin (TcnA). The amino acid sequence of TpeL is shorter than these toxins, and the homologous region was located at the N-terminal site. Eighteen strains of C. perfringens types A, B and C were surveyed for the presence of the tpeL gene by PCR. The tpeL gene was detected in all type B (one strain) and C strains (five strains), but not in any type A strains (12 strains). TpeL was detected in culture filtrates of the five type C strains by dot-blot analysis, but not in the type B strain. It was concluded that TpeL is a novel toxin similar to the known large clostridial cytotoxins. Furthermore, the data indicated that TpeL is produced by many C. perfringens type C strains.

  6. Boron Nitride Nanoribbons from Exfoliation of Boron Nitride Nanotubes

    NASA Technical Reports Server (NTRS)

    Hung, Ching-Cheh; Hurst, Janet; Santiago, Diana

    2017-01-01

    Two types of boron nitride nanotubes (BNNTs) were exfoliated into boron nitride nanoribbons (BNNR), which were identified using transmission electron microscopy: (1) commercial BNNTs with thin tube walls and small diameters. Tube unzipping was indicated by a large decrease of the sample's surface area and volume for pores less than 2 nm in diameter. (2) BNNTs with large diameters and thick walls synthesized at NASA Glenn Research Center. Here, tube unraveling was indicated by a large increase in external surface area and pore volume. For both, the exfoliation process was similar to the previous reported method to exfoliate commercial hexagonal boron nitride (hBN): Mixtures of BNNT, FeCl3, and NaF (or KF) were sequentially treated in 250 to 350 C nitrogen for intercalation, 500 to 750 C air for exfoliation, and finally HCl for purification. Property changes of the nanosized boron nitride throughout this process were also similar to the previously observed changes of commercial hBN during the exfoliation process: Both crystal structure (x-ray diffraction data) and chemical properties (Fourier-transform infrared spectroscopy data) of the original reactant changed after intercalation and exfoliation, but most (not all) of these changes revert back to those of the reactant once the final, purified products are obtained.

  7. Is Botulinum Toxin Type A a Valuable Adjunct During Femoral Lengthening? A Randomized Trial.

    PubMed

    Park, Hoon; Shin, Soowan; Shin, Han Sol; Kim, Hyun Woo; Kim, Dong Wook; Lee, Dong Hoon

    2016-12-01

    Reduced joint ROM and distraction-induced pain are common complaints of patients who have undergone gradual femoral lengthening. Attempts to reduce the effects of lengthening on joint motion have included the use of botulinum toxin to reduce the muscle forces that restrict motion. The benefits of this approach during femoral lengthening, however, have not been conclusively established. We wished to evaluate the effects of botulinum toxin type A (BtX-A) injection in the anterior thigh muscles during femoral distraction osteogenesis on adjacent joint ROM and distraction-induced pain. We asked: (1) Does injection of BtX-A in the quadriceps muscles lead to improved knee and hip motion during femoral lengthening? (2) Does injection of BtX-A reduce pain during femoral lengthening? A single-center, double-blind, randomized placebo-controlled trial was conducted. Forty-four patients (88 femurs) undergoing bilateral femoral lengthening for familial short stature were included in the study. BtX-A (200 IU) was injected intraoperatively in the quadriceps muscles of one thigh. An equal volume of sterile normal saline was injected in the other thigh as a control. Selection of the limb receiving the toxin was randomized. Clinical evaluation included a VAS score for pain measurement, ROM evaluation of the hips and knees, and measurement of thigh circumference. Side-to-side differences were analyzed throughout the entire consolidation phase. No patients were lost to followup, leaving 44 patients (88 femurs). The mean followup was 26 months (range, 14-40 months). The distraction rate and final length of gain were similar between treated and control limbs. A priori power analysis suggested that 44 legs were required in each group to achieve statistical significance of 0.05 with 90% power to detect a 50% difference in treatment effect between treatment and control groups. There were no differences in hip ROM, knee ROM, or maximal thigh circumference between the two lower extremities

  8. Directed Therapy for Exfoliation Syndrome

    PubMed Central

    Angelilli, Allison; Ritch, Robert

    2009-01-01

    Exfoliation syndrome (XFS) is an age-related disorder of the extracellular matrix that leads the production of abnormal fibrillar material that leads to elevated intraocular pressure and a relatively severe glaucoma. Exfoliation material is deposited in numerous ocular tissues and extraocular organs. XFS is associated with ocular ischemia, cerebrovascular disease, neurodegenerative disease and cardiovascular disease. Current modalities of treatment include intraocular pressure lowering with topical antihypertensives, laser trabeculoplasty and filtration surgery. The disease paradigm for XFS should be expanded to include directed therapy designed specifically to target the underlying disease process. Potential targets include preventing the formation or promoting the depolymerization of exfoliation material. Novel therapies targeting trabecular meshwork may prove particularly useful in the care of exfoliative glaucoma. The systemic and ocular associations of XFS underscore the need for a comprehensive search for neuroprotective agents in its treatment. PMID:19888433

  9. Botulinum toxin type A reduces pain supersensitivity in experimental diabetic neuropathy: bilateral effect after unilateral injection.

    PubMed

    Bach-Rojecky, Lidija; Salković-Petrisić, Melita; Lacković, Zdravko

    2010-05-10

    We investigated antinociceptive activity of botulinum toxin type A (BTX-A) in a model of diabetic neuropathic pain in rats. Male Wistar rats were made diabetic by a single intraperitoneal injection of streptozotocin (80mg/kg). Sensitivity to mechanical and thermal stimuli was measured with the paw-pressure and hot-plate test, respectively. The formalin test was used to measure sensitivity to chemical stimuli. Diabetic animals with pain thresholds lower for at least 25% compared to the non-diabetic group were considered neuropathic and were injected with BTX-A either subcutaneously (3, 5 and 7U/kg) or intrathecally (1U/kg). Mechanical and thermal sensitivity was measured at several time-points. After peripheral application, BTX-A (5 and 7U/kg) reduced mechanical and thermal hypersensitivity not only on ipsilateral, but on contralateral side, too. The antinociceptive effect started 5days following BTX-A injection and lasted at least 15days. Formalin-induced hypersensitivity in diabetic animals was abolished as well. When applied intrathecally, BTX-A (1U/kg) reduced diabetic hyperalgesia within 24h supporting the assumption of retrograde axonal transport of BTX-A from the peripheral site of injection to central nervous system. The results presented here demonstrate the long-lasting pain reduction after single BTX-A injection in the animals with diabetic neuropathy. The bilateral pain reduction after unilateral toxin application and the effectiveness of lower dose with the faster onset after the intrathecal injection suggest the involvement of the central nervous system in the antinociceptive action of BTX-A in painful diabetic neuropathy. Copyright 2010 Elsevier B.V. All rights reserved.

  10. Improved traceability of Shiga-toxin-producing Escherichia coli using CRISPRs for detection and typing.

    PubMed

    Delannoy, Sabine; Beutin, Lothar; Fach, Patrick

    2016-05-01

    Among strains of Shiga-toxin-producing Escherichia coli (STEC), seven serogroups (O26, O45, O103, O111, O121, O145, and O157) are frequently associated with severe clinical illness in humans. The development of methods for their reliable detection from complex samples such as food has been challenging thus far, and is currently based on the PCR detection of the major virulence genes stx1, stx2, and eae, and O-serogroup-specific genes. However, this approach lacks resolution. Moreover, new STEC serotypes are continuously emerging worldwide. For example, in May 2011, strains belonging to the hitherto rarely detected STEC serotype O104:H4 were identified as causative agents of one of the world's largest outbreak of disease with a high incidence of hemorrhagic colitis and hemolytic uremic syndrome in the infected patients. Discriminant typing of pathogens is crucial for epidemiological surveillance and investigations of outbreaks, and especially for tracking and tracing in case of accidental and deliberate contamination of food and water samples. Clustered regularly interspaced short palindromic repeats (CRISPRs) are composed of short, highly conserved DNA repeats separated by unique sequences of similar length. This distinctive sequence signature of CRISPRs can be used for strain typing in several bacterial species including STEC. This review discusses how CRISPRs have recently been used for STEC identification and typing.

  11. Thymoma-associated exfoliative dermatitis in cats.

    PubMed

    Rottenberg, S; von Tscharner, C; Roosje, P J

    2004-07-01

    Five cases of exfoliative dermatitis in cats were presented from 1996 to 2002 in which a feline thymoma was found by postmortem or postsurgical examination. Besides abundant exfoliation of keratin squames and layers, the histologic picture of the skin revealed a similar pattern of interface dermatitis with predominantly CD3+ lymphocytes and fewer mast cells and plasma cells. In the epidermal basal layer a hydropic degeneration of keratinocytes was present. In all cases an infundibular lymphocytic mural folliculitis and absence of or drastic decrease in the number of sebaceous glands occurred. In addition to the so far described cell-poor type, we also found examples of a cell-rich skin lesion. Together with the clinical observation of generalized exfoliative dermatitis, the histologic pattern of this dermatitis was suggestive of an underlying thymoma. The pathogenesis of this skin disease in association with thymic neoplasia remains obscure, and our results contradict the hypothesis of production of autoantibodies that cross-react with epithelial antigens. The morphology of the thymomas and CD3 expression of the thymocytes varied and did not seem to have an impact on the dermal lesions.

  12. Utilization of quail and chicken embryos for the detection of botulinum toxin type A activity

    USDA-ARS?s Scientific Manuscript database

    Clostridium botulinum is a ubiquitous microorganism which can produce botulinum toxins and the ability to assess toxin activity in a food sample is critical. As an alternative to the mouse assay incubating quail (Coturnix coturnix japonica) and chicken (Gallus gallus domestics) embryos were evaluat...

  13. Evaluation of a quali embryo model for the detection of botulism toxin type A activity

    USDA-ARS?s Scientific Manuscript database

    The Japanese quail embryo (Coturnix japonica) was evaluated for use as a bioassay to detect biologically active botulinum toxin serotype A (BoNT/A). Day 15 of incubation embryos were injected with decreasing dosages of BoNT/A from 250 to 0.5 ng of toxin. At 1 day post-injection, embryos receiving ...

  14. Mapping of the continuous epitopes displayed on the Clostridium perfringens type D epsilon-toxin.

    PubMed

    Alves, Guilherme Guerra; Machado-de-Ávila, Ricardo Andrez; Chávez-Olórtegui, Carlos Delfin; Silva, Rodrigo Otávio Silveira; Lobato, Francisco Carlos Faria

    2017-02-03

    The epsilon toxin, produced by Clostridium perfringens, is responsible for enterotoxemia in ruminants and is a potential bioterrorism agent. In the present study, 15 regions of the toxin were recognized by antibodies present in the serum, with different immunodominance scales, and may be antigen determinants that can be used to formulate subunit vaccines.

  15. Evaluation of a quail embryo model for the detection of botulinum toxin type A activity

    USDA-ARS?s Scientific Manuscript database

    The quail embryo was evaluated for use as a bioassay to detect biologically active botulinum toxin serotype A (BoNT/A). Day 15 of incubation embryos were injected with decreasing dosages of BoNT/A from 250 to 0.5 ng of toxin. At 1 day post-injection, embryos receiving 20 ng of BoNT or higher had m...

  16. Prevalence of Staphylococcus aureus toxins and nasal carriage in furuncles and impetigo.

    PubMed

    Durupt, F; Mayor, L; Bes, M; Reverdy, M-E; Vandenesch, F; Thomas, L; Etienne, J

    2007-12-01

    The precise role of Staphylococcus aureus toxins and nasal carriage in common skin infections remains unclear. To seek correlations between toxin expression, S. aureus nasal carriage and clinical manifestations in patients with community-acquired furuncles and impetigo. From November 2004 to August 2005, we studied clinical data and bacteriological samples prospectively collected from 121 patients presenting with furuncles or impetigo. Sixty-four patients (31 with furuncles and 33 with impetigo) had S. aureus-positive skin culture. Panton-Valentine leukocidin (PVL) genes were present in 13 of 31 (42%) isolates from furuncles and were associated with epidemic furunculosis. Exfoliative toxin genes were present in 10 of 10 (100%) and 12 of 21 (57%) bullous and nonbullous impetigo isolates, respectively. Nasal carriage of S. aureus was found in 58% of patients overall. It was strongly associated with chronic furunculosis but not with simple furuncles (88% vs. 29%, P < 0.007). Skin and nose isolates from a given patient always had identical characteristics. Methicillin-resistant S. aureus accounted for four of 64 (6%) positive skin cultures. PVL is not involved in all types of furuncles but is associated with epidemic furunculosis. Both bullous and nonbullous forms of impetigo are associated with exfoliative toxins. Staphylococcus aureus nasal carriage is associated with the chronicity of furuncles.

  17. Antinociceptive effect of botulinum toxin type A on experimental abdominal pain.

    PubMed

    Drinovac, Višnja; Bach-Rojecky, Lidija; Babić, Ana; Lacković, Zdravko

    2014-12-15

    Visceral pain, especially in the abdominal region, represents one of the most common types of pain. Its chronic form is usually very hard to treat by conventional analgesic agents and adjuvants. We investigated the antinociceptive effect of botulinum toxin type A (BTX-A) in male Wistar rats in two models of visceral pain: peritonitis induced by intraperitoneal injection of 1% acetic acid and colitis induced by intracolonic instillation of 0.1% capsaicin. Pain was measured as the number of abdominal writhes. Additionally, referred mechanical sensitivity in the ventral abdominal area was evaluated by von Frey test and the extent of spinal c-Fos expression was immunohistochemically examined. BTX-A significantly reduced the number of abdominal writhes in both models of visceral pain after intrathecal application in a dose of 2 U/kg. In the experimental colitis model, BTX-A (2 U/kg) reduced both referred mechanical allodynia and c-Fos expression in the dorsal horn of the spinal cord (S2/S3 segments). In contrast to intrathecal administration, BTX-A (2 U/kg) administered into the cisterna magna had no effect on pain suggesting that the primary site of its action is a spinal cord.

  18. Botulinum toxin type A reduces hyperalgesia and TRPV1 expression in rats with neuropathic pain.

    PubMed

    Xiao, Lizu; Cheng, Jianguo; Zhuang, Yu; Qu, Wenchun; Muir, Jeffery; Liang, Haowen; Zhang, Deren

    2013-02-01

    We aim to determine the effects of botulinum toxin type A (BTX-A) on the thresholds of pain and the expression of transient receptor potential vanilloid type 1 (TRPV1) in the dorsal root ganglion (DRG) in rats with neuropathic pain induced by selective ventral root transection (VRT). Neuropathic pain was induced by transection of the lumbar 5 ventral root in male Sprague-Dawley rats. BTX-A or saline was administered to the plantar surface by subcutaneous injection. SB366791 (an inhibitor of TRPV1) was administered intraperitoneally. Behavioral tests were conducted preoperatively and at predefined postoperative days. The expression of TRPV1 was detected and quantified by immunohistochemistry and Western blotting at postoperative days 3, 7, 14, and 21. TRPV1 expression increased significantly in the L4 ∼5 dorsal root ganglia 7 days after L5 VRT compared with the sham-operated control (P < 0.05). This increase persisted for at least 21 days. The thresholds of foot withdrawal to mechanical and thermal stimulation decreased significantly as well. Subcutaneous injection of BTX-A significantly and dose-dependently reduced the expression of TRPV1 (P < 0.05) and partially reversed the pain thresholds. Upregulation of TRPV1 expression in the DRG is an important mechanism of neuropathic pain induced by the VRT. The analgesic effect of BTX-A is most likely mediated through reduction of TRPV1 expression in the nociceptors. Wiley Periodicals, Inc.

  19. Type II toxin-antitoxin systems are unevenly distributed among Escherichia coli phylogroups.

    PubMed

    Fiedoruk, Krzysztof; Daniluk, Tamara; Swiecicka, Izabela; Sciepuk, Malgorzata; Leszczynska, Katarzyna

    2015-01-01

    Type II toxin-antitoxin systems (TAs) are bicistronic operons ubiquitous in prokaryotic genomes, displaying multilevel association with cell physiology. Various possible functions have been assigned to TAs, ranging from beneficial for their hosts, such as a stress response, dormancy and protection against genomic parasites, to detrimental or useless functions, such as selfish alleles. As there is a link between several Escherichia coli features (e.g. virulence, lifestyle) and the phylogeny of this species, we hypothesized a similar association with TAs. Using PCR we studied the distribution of 15 chromosomal and plasmidic type II TA loci in 84 clinical E. coli isolates in relation to their main phylogenetic groups (A, B1, B2 and D). In addition, we performed in silico searching of these TA loci in 60 completely sequenced E. coli genomes deposited in GenBank. The highest number of TA loci per strain was observed in group A (mean 8.2, range 5-12) and the lowest in group B2 (mean 4.2, range 2-8). Moreover, significant differences in the prevalence of nine chromosomal TAs among E. coli phylogroups were noted. In conclusion, the presence of some chromosomal TAs in E. coli is phylogroup-related rather than a universal feature of the species. In addition, their limited collection in group B2 clearly distinguish it from the other E. coli phylogroups.

  20. Enhanced expression of recombinant beta toxin of Clostridium perfringens type B using a commercially available Escherichia coli strain.

    PubMed

    Bakhshi, Fatemah; Pilehchian Langroudi, Reza; Eimani, Bahram Golestani

    2016-06-30

    Clostridium perfringens beta toxin is only produced by types B and C and plays an important role in many human and animal diseases, causing fatal conditions that originate in the intestines. We compared the expression of C. perfringens type B vaccine strain recombinant beta toxin gene in the Escherichia coli strains Rosetta(DE3) and BL21(DE3). The beta toxin gene was extracted from pJETβ and ligated with pET22b(+). pET22β was transformed into E. coli strains BL21(DE3) and Rosetta(DE3). Recombinant protein was expressed as a soluble protein after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction in strain Rosetta(DE3) but not in BL21(DE3). Expression was optimised by growing recombinant cells at 37 °C and at an induction of 0.5 mM, 1 mM, 1.5 mM IPTG. Expression was evaluated using sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE). The recombinant protein was purified via Ni-NTA and was analysed using western blot. We concluded that E. coli strain RosettaTM(DE3) can enhance the expression of C. perfringens recombinant beta toxin.

  1. [Botulinum toxin type A for the treatment of spasticity in children with cerebral palsy].

    PubMed

    Dimitrijević, Lidija; Stanković, Ivona; Zivković, Vesna; Mikov, Aleksandra; Colović, Hristina; Janković, Irena

    2007-08-01

    Cerebral palsy (CP) is the most common physical disability in childhood. Children have problems with motor functions as a result of limbs spasticity, which leads to severe contractures and limbs deformity. There is a growing interest in the therapeutic role of botulinum toxin type A (BTA) in CP. The aim of this study was to examine the effects of BTA on spasticity, active range of motion and functional motor outcomes in children with CP. This study included 42 children of both sexes, aged 2-6 years, with spastic CP, divided into two groups: group I (21 child) treated with BTA and physical therapy, and group II (21 child) treated with physical therapy only. The following parameters were analyzed: spasticity; active range of motion of the hip, knee and ankle, and functional motor outcome. These parameters measurements were carried out four times in both groups: before the treatment, three, eight and 16 weeks after the beginning of the treatment. The obtained results were statistically processed and compared. There was no evidence of any significant difference between the groups before the treatment. After eight weeks there was a remarkable difference concerning spasticity reducing on behalf of the group I (group I -- 0.76 +/- 0.51 vs. II group -- 2.17 +/- 0.64; p < 0.0001). There was statistically significant difference concerning active range of motion increasing on behalf of the group I (hip abduction: group I --44.37 +/- 1.13(0) vs. group II -- 32.61 +/- 8.07(0),p < 0,01; knee extension: group I -- 0.77 +/- 1.82(0) vs. II group -- 14.99 +/- 7.61(0), p < 0.01; dorsiflexion of the foot: group I -- 11.50 +/- 6.08(0) vs. group II -- 8.98 +/- 7.85(0), p < 0.01). A statistically significant difference was found after 16 weeks in functional motor outcome as well, on behalf of the group I: functional motor abilities level in the group I was 1.86 vs. 2.71 in the group II, p < 0.05. Botulinum toxin type A application leads to an important spasticity decreasing, active range

  2. Detection, Characterization, and Typing of Shiga Toxin-Producing Escherichia coli.

    PubMed

    Parsons, Brendon D; Zelyas, Nathan; Berenger, Byron M; Chui, Linda

    2016-01-01

    Shiga toxin-producing Escherichia coli (STEC) are responsible for gastrointestinal diseases reported in numerous outbreaks around the world. Given the public health importance of STEC, effective detection, characterization and typing is critical to any medical laboratory system. While non-O157 serotypes account for the majority of STEC infections, frontline microbiology laboratories may only screen for STEC using O157-specific agar-based methods. As a result, non-O157 STEC infections are significantly under-reported. This review discusses recent advances on the detection, characterization and typing of STEC with emphasis on work performed at the Alberta Provincial Laboratory for Public Health (ProvLab). Candidates for the detection of all STEC serotypes include chromogenic agars, enzyme immunoassays (EIA) and quantitative real time polymerase chain reaction (qPCR). Culture methods allow further characterization of isolates, whereas qPCR provides the greatest sensitivity and specificity, followed by EIA. The virulence gene profiles using PCR arrays and stx gene subtypes can subsequently be determined. Different non-O157 serotypes exhibit markedly different virulence gene profiles and a greater prevalence of stx1 than stx2 subtypes compared to O157:H7 isolates. Finally, recent innovations in whole genome sequencing (WGS) have allowed it to emerge as a candidate for the characterization and typing of STEC in diagnostic surveillance isolates. Methods of whole genome analysis such as single nucleotide polymorphisms and k-mer analysis are concordant with epidemiological data and standard typing methods, such as pulsed-field gel electrophoresis and multiple-locus variable number tandem repeat analysis while offering additional strain differentiation. Together these findings highlight improved strategies for STEC detection using currently available systems and the development of novel approaches for future surveillance.

  3. Detection, Characterization, and Typing of Shiga Toxin-Producing Escherichia coli

    PubMed Central

    Parsons, Brendon D.; Zelyas, Nathan; Berenger, Byron M.; Chui, Linda

    2016-01-01

    Shiga toxin-producing Escherichia coli (STEC) are responsible for gastrointestinal diseases reported in numerous outbreaks around the world. Given the public health importance of STEC, effective detection, characterization and typing is critical to any medical laboratory system. While non-O157 serotypes account for the majority of STEC infections, frontline microbiology laboratories may only screen for STEC using O157-specific agar-based methods. As a result, non-O157 STEC infections are significantly under-reported. This review discusses recent advances on the detection, characterization and typing of STEC with emphasis on work performed at the Alberta Provincial Laboratory for Public Health (ProvLab). Candidates for the detection of all STEC serotypes include chromogenic agars, enzyme immunoassays (EIA) and quantitative real time polymerase chain reaction (qPCR). Culture methods allow further characterization of isolates, whereas qPCR provides the greatest sensitivity and specificity, followed by EIA. The virulence gene profiles using PCR arrays and stx gene subtypes can subsequently be determined. Different non-O157 serotypes exhibit markedly different virulence gene profiles and a greater prevalence of stx1 than stx2 subtypes compared to O157:H7 isolates. Finally, recent innovations in whole genome sequencing (WGS) have allowed it to emerge as a candidate for the characterization and typing of STEC in diagnostic surveillance isolates. Methods of whole genome analysis such as single nucleotide polymorphisms and k-mer analysis are concordant with epidemiological data and standard typing methods, such as pulsed-field gel electrophoresis and multiple-locus variable number tandem repeat analysis while offering additional strain differentiation. Together these findings highlight improved strategies for STEC detection using currently available systems and the development of novel approaches for future surveillance. PMID:27148176

  4. Lysogeny with Shiga Toxin 2-Encoding Bacteriophages Represses Type III Secretion in Enterohemorrhagic Escherichia coli

    PubMed Central

    Xu, Xuefang; McAteer, Sean P.; Tree, Jai J.; Shaw, Darren J.; Wolfson, Eliza B. K.; Beatson, Scott A.; Roe, Andrew J.; Allison, Lesley J.; Chase-Topping, Margo E.; Mahajan, Arvind; Tozzoli, Rosangela; Woolhouse, Mark E. J.; Morabito, Stefano; Gally, David L.

    2012-01-01

    Lytic or lysogenic infections by bacteriophages drive the evolution of enteric bacteria. Enterohemorrhagic Escherichia coli (EHEC) have recently emerged as a significant zoonotic infection of humans with the main serotypes carried by ruminants. Typical EHEC strains are defined by the expression of a type III secretion (T3S) system, the production of Shiga toxins (Stx) and association with specific clinical symptoms. The genes for Stx are present on lambdoid bacteriophages integrated into the E. coli genome. Phage type (PT) 21/28 is the most prevalent strain type linked with human EHEC infections in the United Kingdom and is more likely to be associated with cattle shedding high levels of the organism than PT32 strains. In this study we have demonstrated that the majority (90%) of PT 21/28 strains contain both Stx2 and Stx2c phages, irrespective of source. This is in contrast to PT 32 strains for which only a minority of strains contain both Stx2 and 2c phages (28%). PT21/28 strains had a lower median level of T3S compared to PT32 strains and so the relationship between Stx phage lysogeny and T3S was investigated. Deletion of Stx2 phages from EHEC strains increased the level of T3S whereas lysogeny decreased T3S. This regulation was confirmed in an E. coli K12 background transduced with a marked Stx2 phage followed by measurement of a T3S reporter controlled by induced levels of the LEE-encoded regulator (Ler). The presence of an integrated Stx2 phage was shown to repress Ler induction of LEE1 and this regulation involved the CII phage regulator. This repression could be relieved by ectopic expression of a cognate CI regulator. A model is proposed in which Stx2-encoding bacteriophages regulate T3S to co-ordinate epithelial cell colonisation that is promoted by Stx and secreted effector proteins. PMID:22615557

  5. The gene for type A streptococcal exotoxin (erythrogenic toxin) is located in bacteriophage T12.

    PubMed Central

    Weeks, C R; Ferretti, J J

    1984-01-01

    The infection of Streptococcus pyogenes T25(3) with the temperate bacteriophage T12 results in the conversion of the nontoxigenic strain to type A streptococcal exotoxin (erythrogenic toxin) production. Although previous research has established that integration of the bacteriophage genome into the host chromosome is not essential for exotoxin production, the location of the gene on the bacteriophage or bacterial chromosome had not been determined. In the present investigation, recombinant DNA techniques were used to determine whether the gene specifying type A streptococcal exotoxin (speA) production is located on the bacteriophage chromosome. Bacteriophage T12 was obtained from S. pyogenes T25(3)(T12) by induction with mitomycin C, and after isolation of bacteriophage DNA by phenol-chloroform extraction, the DNA was digested with restriction enzymes and ligated with Escherichia coli plasmid pHP34 or the Streptococcus-E. coli shuttle vector pSA3. Transformation of E. coli HB101 with the recombinant molecules allowed selection of E. coli clones containing bacteriophage T12 genes. Immunological assays with specific antibody revealed the presence of type A streptococcal exotoxin in sonicates of E. coli transformants. Subcloning experiments localized the speA gene to a 1.7-kilobase segment of the bacteriophage T12 genome flanked by SalI and HindIII sites. Introduction of the pSA3 vector containing the speA gene into Streptococcus sanguis (Challis) resulted in transformants that secreted the type A exotoxin. Immunological analysis showed that the type A streptococcal exotoxin produced by E. coli and S. sanguis transformants was identical to the type A exotoxin produced by S. pyogenes T25(3)(T12). Southern blot hybridizations with the cloned fragment confirmed its presence in the bacteriophage T12 genome and its absence in the T25(3) nonlysogen. Therefore, the gene for type A streptococcal exotoxin is located in the bacteriophage genome, and conversion of S. pyogenes T

  6. Resiniferatoxin and botulinum toxin type A for treatment of lower urinary tract symptoms.

    PubMed

    Cruz, Francisco; Dinis, Paulo

    2007-10-01

    Resiniferatoxin (RTX) and botulinum toxin subtype A (BTX-A) are increasingly viewed as potential treatments for lower urinary tract symptoms (LUTS) refractory to conventional therapy. RTX, a capsaicin analogue devoid of severe pungent properties, acts by desensitizing the transient receptor potential vanilloid type 1 (TRPV1) receptor and inactivating C-fibers. BTX-A cleaves soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins in afferent and efferent nerve endings, therefore impeding the fusion of synaptic vesicles with the neuronal membrane necessary for the release of neurotransmitters. In patients with neurogenic and idiopathic detrusor overactivity, RTX and BTX-A have been shown to increase the volume to first detrusor contraction, increase bladder capacity, and improve urinary incontinence and quality of life. Recent data also suggest a role for these neurotoxins in treating urgency, the primary symptom in overactive bladder (OAB) syndrome. Furthermore, experimental data strongly support the use of both neurotoxins in the treatment of pain and frequency in patients with interstitial cystitis/painful bladder syndrome (IC/PBS), although the results from available clinical trials for this use are still inconclusive. In spite of promising results overall, it should be made clear that the administration of these neurotoxins is still considered an experimental procedure and that more clinical studies are necessary before a license for their use will be issued by health authorities.

  7. Motor function following multilevel botulinum toxin type A treatment in children with cerebral palsy.

    PubMed

    Desloovere, Kaat; Molenaers, Guy; De Cat, Jos; Pauwels, Petra; Van Campenhout, Anja; Ortibus, Els; Fabry, Guy; De Cock, Paul

    2007-01-01

    This study evaluated the effects of multilevel botulinum toxin type A (BTX-A) treatments on the gait pattern of children with spastic cerebral palsy (Gross Motor Function Classification System Levels I-III). In this nested case-control design, 30 children (mean age 6y 11mo [SD 1y 5mo]; 21 males, nine females; 19 with hemiplegia, 11 with diplegia) were treated according to best practice guidelines in paediatric orthopaedics, including BTX-A injections. A matched control group of 30 children (mean age 7y 8mo [SD 1y 10mo]; 13 males, 17 females; 19 with hemiplegia, 11 with diplegia) were treated identically, but without BTX-A. Motor development status at 5 to 10 years of age was assessed by means of three-dimensional gait analysis at a mean time of 1 year 10 months (SD 10mo) after the last BTX-A treatment. The control group showed a significantly more pronounced pathological gait pattern than the BTX-A group. Major differences were found for pelvic anterior tilt, maximum hip and knee extension, and internal hip rotation. These results provide evidence for a prolonged effect of BTX-A and suggest that BTX-A injections, in combination with common conservative treatment options, result in a gait pattern that is less defined by secondary problems (e.g. bony deformities) at 5 to 10 years of age, minimizing the need for complex surgery at a later age and enhancing quality of life.

  8. Auto-Assembling Detoxified Staphylococcus aureus Alpha-Hemolysin Mimicking the Wild-Type Cytolytic Toxin

    PubMed Central

    Fiaschi, Luigi; Di Palo, Benedetta; Scarselli, Maria; Pozzi, Clarissa; Tomaszewski, Kelly; Galletti, Bruno; Nardi-Dei, Vincenzo; Arcidiacono, Letizia; Mishra, Ravi P. N.; Mori, Elena; Pallaoro, Michele; Falugi, Fabiana; Torre, Antonina; Fontana, Maria Rita; Soriani, Marco; Bubeck Wardenburg, Juliane; Grandi, Guido; Rappuoli, Rino

    2016-01-01

    Staphylococcus aureus alpha-hemolysin (Hla) assembles into heptameric pores on the host cell membrane, causing lysis, apoptosis, and junction disruption. Herein, we present the design of a newly engineered S. aureus alpha-toxin, HlaPSGS, which lacks the predicted membrane-spanning stem domain. This protein is able to form heptamers in aqueous solution in the absence of lipophilic substrata, and its structure, obtained by transmission electron microscopy and single-particle reconstruction analysis, resembles the cap of the wild-type cytolytic Hla pore. HlaPSGS was found to be impaired in binding to host cells and to its receptor ADAM10 and to lack hemolytic and cytotoxic activity. Immunological studies using human sera as well as sera from mice convalescent from S. aureus infection suggested that the heptameric conformation of HlaPSGS mimics epitopes exposed by the cytolytic Hla pore during infection. Finally, immunization with this newly engineered Hla generated high protective immunity against staphylococcal infection in mice. Overall, this study provides unprecedented data on the natural immune response against Hla and suggests that the heptameric HlaPSGS is a highly valuable vaccine candidate against S. aureus. PMID:27030589

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

    PubMed

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

    2011-02-01

    Over the past decade, facial cosmetic procedures have become more commonplace in 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 dynamic rhytids and lines with BoNTA is effective and produces high rates of improvement with rapid onset and long duration of action (longer than 4 months for some patients) compared with placebo. This paper considers the history and pharmacology of this neurotoxin, and focusses on the literature concerning the treatment of different facial areas with BoNTA. It also presents clinical guidelines on the treatment of glabellar lines, the frontalis muscle, peri-orbital lines, gummy smile and masseter muscle hypertrophy. Knowledge about the mechanisms of action and the ability to use BoNTA as an adjunctive treatment are mandatory for those working in the field of cosmetic facial surgery. Copyright © 2010 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  10. Use of botulinum toxin type A in symptomatic accessory soleus muscle: first five cases.

    PubMed

    Isner-Horobeti, M-E; Muff, G; Lonsdorfer-Wolf, E; Deffinis, C; Masat, J; Favret, F; Dufour, S P; Lecocq, J

    2016-11-01

    Symptomatic accessory soleus muscle (ASM) can cause exercise-induced leg pain due to local nerve/vascular compression, muscle spasm, or local compartment syndrome. As intramuscular injections of botulinum toxin type A (BTX-A) can reduce muscle tone and mass, we investigated whether local BTX-A injections relieve the pain associated with symptomatic ASM. We describe five patients presenting peri/retromalleolar exertional pain and a contractile muscle mass in the painful region. Com-pression neuropathy was ruled out by electromyo-graphic analysis of the lower limb muscles. Doppler ultrasonography was normal, excluding a local vascular compression. ASM was confirmed by magnetic resonance imaging. After a treadmill stress test, abnormal intramuscular pressure values in the ASM, confirmed the diagnosis of compartment syndrome only in one patient. All five patients received BTX-A injections in two points of the ASM. The treatment efficacy was evaluated based on the disappearance of exercise-induced pain and the resumption of normal physical and sports activities. After BTX-A injection, exertional pain disappeared and all five patients resumed their normal level of physical and sports performances. Neither side effects nor motor deficits were observed. BTX-A is well tolerated in patients with ASM and could be used as a new conservative therapeutic strategy for the treatment of symptomatic ASM before surgery.

  11. Knee flexion contracture treated with botulinum toxin type A in patients with haemophilia (PWH).

    PubMed

    Daffunchio, C; Caviglia, H; Nassif, J; Morettil, N; Galatro, G

    2016-01-01

    Knee flexion contracture (KFC) remains a common complication of haemoarthrosis in children and young adults with haemophilia. If the KFC is not treated properly it produces disability, postural and gait abnormalities. Evaluate the effectiveness of conservative treatment of KFC with Botulinum toxin type A (BTX-A) in PWH. Seventeen patients were treated, with 21 affected knees. Mean age was 26 years. The mean follow up was 12 months. We evaluated flexion and KFC pretreatment BTX-A and up to 12 months posttreatment. BTX-A application was in hamstring and calf muscles. To evaluate the function, a questionnaire about different activities was made, and it was checked 3, 6 and 12 months after BTX-A. According to the degree of KFC, knees were divided into 3 groups: Group 1: -10° to -30° (n = 10), Group 2: -31° to -45° (n = 6) Group 3: -46° or more (n = 5). The average KFC improved from -38° to -24°. The improvement was 14° (P < 0.001). The average KFC improvement was 9° in group 1, 17° in group 2, and 23° in group 3. There was a high correlation between the improvement in KFC and the total score of the questionnaire R = 0.77. Treatment of KFC with BTX-A improves knee-related functional activities, with the advantage of being a low-cost procedure and easy to apply. © 2015 John Wiley & Sons Ltd.

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

    PubMed Central

    Brin, Mitchell F; James, Charmaine; Maltman, John

    2014-01-01

    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. PMID:25336912

  13. [Botulinum toxin type A contribution in the treatment of Raynaud's phenomenon due to systemic sclerosis].

    PubMed

    Serri, J; Legré, R; Veit, V; Guardia, C; Gay, A-M

    2013-12-01

    Raynaud's phenomenon is a vasospastic disorder of the extremities that can lead, in the hands, to pain, disability, ischemic ulcers and digital chronic ischemia. Medical and surgical current treatments are not fully effective while causing side effects. Recent studies have emphasized the value of botulinum toxin type A (BTX A) in the management of primary Raynaud's phenomenon. The originality of Raynaud's syndrome secondary to systemic sclerosis is to combine both arterial vasospasm and sclerosis of the arterial wall, what is supposed to reduce BTX A effects. The purpose of this work is to evaluate BTX A efficiency in patients with Raynaud's phenomenon secondary to systemic sclerosis. We performed a prospective study for 12 months. Patients with severe Raynaud's phenomenon due to systemic sclerosis were injected with BTX A in the two hands. Evolution of ischemic ulcers, QuickDASH Score, O2 partial pressure, pain were measured before and 30 days after injection. We treated 18 patients. Thirty days after injection, we noticed a complete healing of ulcers, QuickDASH Score was improved from 39.4 to 20, as the O2 partial pressure from 16 to 42 mmHg and the pain from VNS from 6/10 to 2/10. BTX A appears to improve significantly Raynaud's phenomenon symptomatology in patients with systemic sclerosis despite the component of arterial sclerosis. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

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

    PubMed Central

    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

  15. Growth effects of botulinum toxin type A injected into masseter muscle on a developing rat mandible.

    PubMed

    Kim, J-Y; Kim, S-T; Cho, S-W; Jung, H-S; Park, K-T; Son, H-K

    2008-10-01

    Botulinum toxin type A (BTX-A) reduces the muscular contractions by temporarily inhibiting the release of acetylcholine at the neuromuscular junction. The purpose of this study was to investigate the effects of the BTX-A injected into the masseter muscle of a developing rat mandible. Four-week-old male (no. 80) Sprague-Dawley rats were divided into four groups: control group, saline group, BTX-A group and baseline control group. Rats of baseline group were sacrificed at 0 day to provide baseline values of the mandibular measurements. The masseter muscle of rats in the saline and the BTX-A group were administered with saline and BTX-A solutions respectively. Experimental animals were sacrificed after 4 weeks. The BTX-A group demonstrated smaller mandibular dimension compared with the other groups (P < 0.05). Their condylar cartilages showed increased apoptosis at the proliferation stage of the reserve zone and masseter muscle fibers demonstrated atrophic changes. The result demonstrated BTX-A influence on inhibitory action of the developing mandible because of apoptosis at the proliferation stage of the reserve zone of the condylar cartilage in developing rat mandible.

  16. Growth effects of botulinum toxin type A injected unilaterally into the masseter muscle of developing rats.

    PubMed

    Park, Chanyoung; Park, Kitae; Kim, Jiyeon

    2015-01-01

    To evaluate the effects of botulinum toxin type A (BTX-A) on mandible skeletal development by inducing muscle hypofunction. Four-week-old Sprague-Dawley rats (n=60) were divided into three groups: Group 1 animals served as controls and were injected with saline; Group 2 animals were injected unilaterally with BTX-A (the contralateral side was injected with saline); and Group 3 animals were injected bilaterally with BTX-A. In Group 2, the saline-injected side was designated the control side (Group 2-1), whereas the BTX-A-injected side was designated the experimental side (Group 2-2). After four weeks, the animals were sacrificed, dry skulls were prepared, and mandibles were measured. In the unilateral group, the experimental side (Group 2-2) had reduced dimensions for all mandible measurements compared with the control side (Group 2-1), suggesting a local effect of BTX-A on mandible growth, likely due to muscle reduction. Localized BTX-A injection induced a change in craniofacial growth, and the skeletal effect was unilateral despite both sides of the mandible functioning as one unit.

  17. A double-blind, randomised, crossover trial of two botulinum toxin type a in patients with spasticity.

    PubMed

    Guarany, Fábio Coelho; Picon, Paulo Dornelles; Guarany, Nicole Ruas; dos Santos, Antonio Cardoso; Chiella, Bianca Paula Mentz; Barone, Carolina Rocha; Fendt, Lúcia Costa Cabral; Schestatsky, Pedro

    2013-01-01

    Botulinum toxin type A (btxA) is one of the main treatment choices for patients with spasticity. Prosigne® a new released botulinum toxin serotype A may have the same effectiveness as Botox® in focal dystonia. However, there are no randomized clinical trials comparing these formulations in spasticity treatment. The aim of our study was to compare the efficacy and safety of Prosigne® with Botox® in the treatment of spasticity. We performed a double-blind, randomized, crossover study consisting of 57 patients with clinically meaningful spasticity. The patients were assessed at baseline, 4 and 12 weeks after Prosigne® or Botox® administration. The main outcomes were changes in the patients' Modified Ashworth Scale (MAS), Functional Independence Measure (FIM) and Pediatric Evaluation of Disability Inventory (PEDI) scores and adverse effects related to the botulinum toxin. Both of the toxins were significantly effective in relieving the level of spasticity in adults and children. There were no significant differences found between the Prosigne® and Botox® treatments regarding their MAS, FIM and PEDI scores. Likewise, the incidence of adverse effects was similar between the two groups. Our results suggest that Prosigne® and Botox® are both efficient and comparable with respect to their efficacy and safety for the three month treatment of spasticity. ClinicalTrials.gov NCT00819065.

  18. A Double-Blind, Randomised, Crossover Trial of Two Botulinum Toxin Type A in Patients with Spasticity

    PubMed Central

    Guarany, Fábio Coelho; Picon, Paulo Dornelles; Guarany, Nicole Ruas; dos Santos, Antonio Cardoso; Chiella, Bianca Paula Mentz; Barone, Carolina Rocha; Fendt, Lúcia Costa Cabral; Schestatsky, Pedro

    2013-01-01

    Background Botulinum toxin type A (btxA) is one of the main treatment choices for patients with spasticity. Prosigne® a new released botulinum toxin serotype A may have the same effectiveness as Botox® in focal dystonia. However, there are no randomized clinical trials comparing these formulations in spasticity treatment. The aim of our study was to compare the efficacy and safety of Prosigne® with Botox® in the treatment of spasticity. Methodology/Principal Findings We performed a double-blind, randomized, crossover study consisting of 57 patients with clinically meaningful spasticity. The patients were assessed at baseline, 4 and 12 weeks after Prosigne® or Botox® administration. The main outcomes were changes in the patients’ Modified Ashworth Scale (MAS), Functional Independence Measure (FIM) and Pediatric Evaluation of Disability Inventory (PEDI) scores and adverse effects related to the botulinum toxin. Both of the toxins were significantly effective in relieving the level of spasticity in adults and children. There were no significant differences found between the Prosigne® and Botox® treatments regarding their MAS, FIM and PEDI scores. Likewise, the incidence of adverse effects was similar between the two groups. Conclusion Our results suggest that Prosigne® and Botox® are both efficient and comparable with respect to their efficacy and safety for the three month treatment of spasticity. Trial Registration ClinicalTrials.gov NCT00819065. PMID:23468866

  19. An Integrative Approach to Computational Modelling of the Gene Regulatory Network Controlling Clostridium botulinum Type A1 Toxin Production

    PubMed Central

    Walshaw, John; Peck, Michael W.; Barker, Gary C.

    2016-01-01

    Clostridium botulinum produces botulinum neurotoxins (BoNTs), highly potent substances responsible for botulism. Currently, mathematical models of C. botulinum growth and toxigenesis are largely aimed at risk assessment and do not include explicit genetic information beyond group level but integrate many component processes, such as signalling, membrane permeability and metabolic activity. In this paper we present a scheme for modelling neurotoxin production in C. botulinum Group I type A1, based on the integration of diverse information coming from experimental results available in the literature. Experiments show that production of BoNTs depends on the growth-phase and is under the control of positive and negative regulatory elements at the intracellular level. Toxins are released as large protein complexes and are associated with non-toxic components. Here, we systematically review and integrate those regulatory elements previously described in the literature for C. botulinum Group I type A1 into a population dynamics model, to build the very first computational model of toxin production at the molecular level. We conduct a validation of our model against several items of published experimental data for different wild type and mutant strains of C. botulinum Group I type A1. The result of this process underscores the potential of mathematical modelling at the cellular level, as a means of creating opportunities in developing new strategies that could be used to prevent botulism; and potentially contribute to improved methods for the production of toxin that is used for therapeutics. PMID:27855161

  20. An Integrative Approach to Computational Modelling of the Gene Regulatory Network Controlling Clostridium botulinum Type A1 Toxin Production.

    PubMed

    Ihekwaba, Adaoha E C; Mura, Ivan; Walshaw, John; Peck, Michael W; Barker, Gary C

    2016-11-01

    Clostridium botulinum produces botulinum neurotoxins (BoNTs), highly potent substances responsible for botulism. Currently, mathematical models of C. botulinum growth and toxigenesis are largely aimed at risk assessment and do not include explicit genetic information beyond group level but integrate many component processes, such as signalling, membrane permeability and metabolic activity. In this paper we present a scheme for modelling neurotoxin production in C. botulinum Group I type A1, based on the integration of diverse information coming from experimental results available in the literature. Experiments show that production of BoNTs depends on the growth-phase and is under the control of positive and negative regulatory elements at the intracellular level. Toxins are released as large protein complexes and are associated with non-toxic components. Here, we systematically review and integrate those regulatory elements previously described in the literature for C. botulinum Group I type A1 into a population dynamics model, to build the very first computational model of toxin production at the molecular level. We conduct a validation of our model against several items of published experimental data for different wild type and mutant strains of C. botulinum Group I type A1. The result of this process underscores the potential of mathematical modelling at the cellular level, as a means of creating opportunities in developing new strategies that could be used to prevent botulism; and potentially contribute to improved methods for the production of toxin that is used for therapeutics.

  1. The effect of Clostridium perfringens type C strain CN3685 and its isogenic beta toxin null mutant in goats.

    PubMed

    Garcia, J P; Beingesser, J; Fisher, D J; Sayeed, S; McClane, B A; Posthaus, H; Uzal, F A

    2012-06-15

    Clostridium perfringens type C is an important cause of enteritis and/or enterocolitis in several animal species, including pigs, sheep, goats, horses and humans. The disease is a classic enterotoxemia and the enteric lesions and associated systemic effects are thought to be caused primarily by beta toxin (CPB), one of two typing toxins produced by C. perfringens type C. This has been demonstrated recently by fulfilling molecular Koch's postulates in rabbits and mice. We present here an experimental study to fulfill these postulates in goats, a natural host of C. perfringens type C disease. Nine healthy male or female Anglo Nubian goat kids were inoculated with the virulent C. perfringens type C wild-type strain CN3685, an isogenic CPB null mutant or a strain where the cpb null mutation had been reversed. Three goats inoculated with the wild-type strain presented abdominal pain, hemorrhagic diarrhea, necrotizing enterocolitis, pulmonary edema, hydropericardium and death within 24h of inoculation. Two goats inoculated with the CPB null mutant and two goats inoculated with sterile culture media (negative controls) remained clinically healthy during 24h after inoculation and no gross or histological abnormalities were observed in the tissues of any of them. Reversal of the null mutation to partially restore CPB production also increased virulence; 2 goats inoculated with this reversed mutant presented clinical and pathological changes similar to those observed in goats inoculated with the wild-type strain, except that spontaneous death was not observed. These results indicate that CPB is required for C. perfringens type C to induce disease in goats, supporting a key role for this toxin in natural C. perfringens type C disease pathogenesis. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. The effect of Clostridium perfringens type C strain CN3685 and its isogenic beta toxin null mutant in goats

    PubMed Central

    Garcia, J. P.; Beingesser, J.; Fisher, D. J.; Sayeed, S.; McClane, B. A.; Posthaus, H.; Uzal, F. A.

    2012-01-01

    Clostridium perfringens type C is an important cause of enteritis and/or enterocolitis in several animal species, including pigs, sheep, goats, horses and humans. The disease is a classic enterotoxemia and the enteric lesions and associated systemic effects are thought to be caused primarily by beta toxin (CPB), one of two typing toxins produced by C. perfringens type C. This has been demonstrated recently by fulfilling molecular Koch’s postulates in rabbits and mice. We present here an experimental study to fulfill these postulates in goats, a natural host of C. perfringens type C disease. Nine healthy male or female Anglo Nubian goat kids were inoculated with the virulent C. perfringens type C wild-type strain CN3685, an isogenic CPB null mutant or a strain where the cpb null mutation had been reversed. Three goats inoculated with the wild-type strain presented abdominal pain, hemorrhagic diarrhea, necrotizing enterocolitis, pulmonary edema, hydropericardium and death within 24 h of inoculation. Two goats inoculated with the CPB null mutant and two goats inoculated with sterile culture media (negative controls) remained clinically healthy during 24 h after inoculation and no gross or histological abnormalities were observed in the tissues of any of them. Reversal of the null mutation to partially restore CPB production also increased virulence; 2 goats inoculated with this reversed mutant presented clinical and pathological changes similar to those observed in goats inoculated with the wild-type strain, except that spontaneous death was not observed. These results indicate that CPB is required for C. perfringens type C to induce disease in goats, supporting a key role for this toxin in natural C. perfringens type C disease pathogenesis. PMID:22296994

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

    SciTech Connect

    Nakamura, Toshio; Tonozuka, Takashi; Kotani, Mao; Obata, Kanae; Oguma, Keiji; Nishikawa, Atsushi

    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.

  4. Botulinum toxin type-A injection to treat patients with intractable anismus unresponsive to simple biofeedback training

    PubMed Central

    Zhang, Yong; Wang, Zhen-Ning; He, Lei; Gao, Ge; Zhai, Qing; Yin, Zhi-Tao; Zeng, Xian-Dong

    2014-01-01

    AIM: To evaluate the efficacy of botulinum toxin type A injection to the puborectalis and external sphincter muscle in the treatment of patients with anismus unresponsive to simple biofeedback training. METHODS: This retrospective study included 31 patients suffering from anismus who were unresponsive to simple biofeedback training. Diagnosis was made by anorectal manometry, balloon expulsion test, surface electromyography of the pelvic floor muscle, and defecography. Patients were given botulinum toxin type A (BTX-A) injection and pelvic floor biofeedback training. Follow-up was conducted before the paper was written. Improvement was evaluated using the chronic constipation scoring system. RESULTS: BTX-A injection combined with pelvic floor biofeedback training achieved success in 24 patients, with 23 maintaining persistent satisfaction during a mean period of 8.4 mo. CONCLUSION: BTX-A injection combined with pelvic floor biofeedback training seems to be successful for intractable anismus. PMID:25253964

  5. Botulinum toxin type-A injection to treat patients with intractable anismus unresponsive to simple biofeedback training.

    PubMed

    Zhang, Yong; Wang, Zhen-Ning; He, Lei; Gao, Ge; Zhai, Qing; Yin, Zhi-Tao; Zeng, Xian-Dong

    2014-09-21

    To evaluate the efficacy of botulinum toxin type A injection to the puborectalis and external sphincter muscle in the treatment of patients with anismus unresponsive to simple biofeedback training. This retrospective study included 31 patients suffering from anismus who were unresponsive to simple biofeedback training. Diagnosis was made by anorectal manometry, balloon expulsion test, surface electromyography of the pelvic floor muscle, and defecography. Patients were given botulinum toxin type A (BTX-A) injection and pelvic floor biofeedback training. Follow-up was conducted before the paper was written. Improvement was evaluated using the chronic constipation scoring system. BTX-A injection combined with pelvic floor biofeedback training achieved success in 24 patients, with 23 maintaining persistent satisfaction during a mean period of 8.4 mo. BTX-A injection combined with pelvic floor biofeedback training seems to be successful for intractable anismus.

  6. Botulinum toxin A is effective to treat tension-type headache caused by hemifacial spasm.

    PubMed

    Mizuma, Atsushi; Nagata, Eiichiro; Yasuda, Takashi; Kouchi, Maiko; Nakayama, Taira; Honma, Kazunari; Tokuoka, Kentaro; Kitagawa, Yasuhisa; Nogawa, Shigeru; Takizawa, Shunya

    2017-10-01

    We examined the relationship between hemifacial spasm (HFS; a form of cranio-cervical dystonia) and chronic primary headache, including tension-type headache (TTH). We also examined whether botulinum toxin A (BoNT/A) therapy for HFS ameliorates concomitant TTH. Fifty-one HFS patients receiving BoNT/A therapy were recruited. Patients' characteristics (including age, gender, chronic headache history, exercise habits, stiff neck, cervical spondylolysis history), stress factors, worsening/new onset of headache associated with HFS, and dose of BoNT/A were examined. We diagnosed headache types according to The International Classification of Headache Disorders, 3rd edition, beta. Numerical Rating Scale (NRS) and Headache Impact Test-6 (HIT-6) scores for headache severity were compared between the 6-week baseline before BoNT/A therapy and 6-week follow-up after BoNT/A therapy. Of 51 patients with HFS, 17 (33.3%) reported worsening or new onset of headache (especially TTH) associated with HFS (Group-S), and 34 were not aware of headache (Group-N). Twelve patients (70.6%) in group-S reported improvement of headache after BoNT/A therapy. NRS (from 7 [5-9] to 0 [0-5], p<0.01) and HIT-6 (from 55 [54-64] to 44 [36-52], p<0.001) scores were significantly improved after BoNT/A therapy. Logistic regression analysis revealed significant interaction between TTH associated with HFS and the presence of stress factors (odds ratio 43.11: 2.95-629.39, p<0.001) and history of chronic headache (odds ratio 28.53: 2.96-275.10, p<0.001). Primary headache, especially TTH, is associated with HFS. BoNT/A therapy for HFS may also be indirectly effective for treatment of TTH. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. The efficiency of botulinum toxin type A for the treatment of masseter muscle pain in patients with temporomandibular joint dysfunction and tension-type headache.

    PubMed

    Pihut, Malgorzata; Ferendiuk, Ewa; Szewczyk, Michal; Kasprzyk, Katarzyna; Wieckiewicz, Mieszko

    2016-01-01

    Temporomandibular joint dysfunction are often accompanied by symptoms of headache such as tension-type headache which is the most frequent spontaneous primary headache. Masseter muscle pain is commonly reported in this group. The purpose of the study was to assess the efficiency of intramuscular botulinum toxin type A injections for treating masseter muscle pain in patients with temporomandibular joint dysfunction and tension-type headache. This prospective outcome study consisted of 42 subjects of both genders aged 19-48 years diagnosed with masseter muscle pain related to temporomandibular joint dysfunction and tension-type headache. The subjects were treated by the intramuscular injection of 21 U (mice units) of botulinum toxin type A (Botox, Allergan) in the area of the greatest cross-section surface of both masseter bellies. Pain intensity was evaluated using visual analogue scale (VAS) and verbal numerical rating scale (VNRS) 1 week before the treatment and 24 weeks after the treatment. The obtained data were analyzed using the Wilcoxon matched pairs test (p ≤ 0,005). The results of this study showed a decrease in the number of referred pain episodes including a decrease in pain in the temporal region bilaterally, a reduction of analgesic drugs intake as well as a decrease in reported values of VAS and VNRS after injections (p = 0,000). The intramuscular botulinum toxin type A injections have been an efficient method of treatment for masseter muscle pain in patients with temporomandibular joint dysfunction and tension-type headache.

  8. Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal epithelial cells and erythrocytes.

    PubMed

    Fujinaga, Yukako; Inoue, Kaoru; Watarai, Shinobu; Sakaguchi, Yoshihiko; Arimitsu, Hideyuki; Lee, Jae-Chul; Jin, Yingji; Matsumura, Takuhiro; Kabumoto, Yuko; Watanabe, Toshihiro; Ohyama, Tohru; Nishikawa, Atsushi; Oguma, Keiji

    2004-05-01

    Clostridium botulinum type C 16S progenitor toxin consists of a neurotoxin (NTX), a non-toxic non-HA (NTNH), and a haemagglutinin (HA). The HA acts as an adhesin, allowing the 16S toxin to bind to intestinal epithelial cells and erythrocytes. In type C, these bindings are dependent on sialic acid. The HA consists of four distinct subcomponents designated HA1, HA2, HA3a and HA3b. To identify the binding subcomponent(s) of HA of type C 16S toxin, all of the HA-subcomponents and some of their precursor forms were produced as recombinant proteins fused to glutathione S-transferase (GST). These proteins were evaluated for their capacity to adhere to intestinal epithelial cells of guinea pig and human erythrocytes. GST-HA1, GST-HA3b and GST-HA3 (a precursor form of HA3a and HA3b) bound intestinal epithelial cells and erythrocytes, whereas GST alone, GST-HA2 and GST-HA3a did not. GST-HA3b and GST-HA3 showed neuraminidase-sensitive binding to the intestinal epithelial cells and erythrocytes, whereas GST-HA1 showed neuraminidase-insensitive binding. TLC binding assay revealed that GST-HA3b and GST-HA3 recognized sialosylparagloboside (SPG) and GM3 in the ganglioside fraction of the erythrocytes, like native type C 16S toxin [Inoue, K. et al. (1999). Microbiology 145, 2533-2542]. On the other hand, GST-HA1 recognized paragloboside (PG; an asialo- derivative of SPG) in addition to SPG and GM3. Deletion mutant analyses of GST-HA3b showed that the C-terminal region of HA3b is important for its binding activity. Based on these data, it is concluded that the HA component contains two distinct carbohydrate-binding subcomponents, HA1 and HA3b, which recognize carbohydrates in different specificities.

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

    ERIC Educational Resources Information Center

    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,…

  10. [Efficacy of semiconductor laser therapy combined with botulinum toxin A type injection in treatment of temporomandibular disorders].

    PubMed

    Jiang, Mei-Yuan; You, Qing-Ling

    2016-12-01

    To observe the efficacy of semiconductor laser therapy in combination with botulinum toxin A type injection in treating temporomandibular disorders and the influence on serum levels of interleukin(IL)-1 and tumor necrosis factor(TNF)-α. Ninety patients with temporomandibular disorders were selected and randomly divided into control group and treatment group with 45 cases in each group. Patients in the control group received semiconductor laser therapy,once per day and 1 course for 1 week. Patients in the treatment group was given semiconductor laser therapy in combination with botulinum toxin A type. 50 U botulinum toxin A type were injected into the masseteric and temporal muscles with 5 injections at different sites in each side. The treatment course of patients was 2 weeks. Fricton scale indexes, pain index, and efficacy were compared between the two groups. Serum levels of IL-1 and TNF-α were detected in both groups. SPSS19.0 software package was used for data analysis. After treatment, PI, DI and CMI in the treatment group were significantly lower than the control group (P<0.01). After 1 and 2 weeks of treatment, VAS score of the treatment group was significantly lower than the control group (P<0.01). The total efficacy rate of the treatment group was 93.33%,which was significantly higher than control group (66.67%, P<0.05). Serum levels of IL-1 and TNF-α of the treatment group were significantly lower than the control group (P<0.01). The efficacy of semiconductor laser therapy in combination with botulinum toxin A type injection in treating temporomandibular disorders is significant,and may be related to decreased serum levels of IL-1 and TNF-α.

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

    ERIC Educational Resources Information Center

    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,…

  12. Comprehensive comparative-genomic analysis of Type 2 toxin-antitoxin systems and related mobile stress response systems in prokaryotes

    PubMed Central

    Makarova, Kira S; Wolf, Yuri I; Koonin, Eugene V

    2009-01-01

    Background The prokaryotic toxin-antitoxin systems (TAS, also referred to as TA loci) are widespread, mobile two-gene modules that can be viewed as selfish genetic elements because they evolved mechanisms to become addictive for replicons and cells in which they reside, but also possess "normal" cellular functions in various forms of stress response and management of prokaryotic population. Several distinct TAS of type 1, where the toxin is a protein and the antitoxin is an antisense RNA, and numerous, unrelated TAS of type 2, in which both the toxin and the antitoxin are proteins, have been experimentally characterized, and it is suspected that many more remain to be identified. Results We report a comprehensive comparative-genomic analysis of Type 2 toxin-antitoxin systems in prokaryotes. Using sensitive methods for distant sequence similarity search, genome context analysis and a new approach for the identification of mobile two-component systems, we identified numerous, previously unnoticed protein families that are homologous to toxins and antitoxins of known type 2 TAS. In addition, we predict 12 new families of toxins and 13 families of antitoxins, and also, predict a TAS or TAS-like activity for several gene modules that were not previously suspected to function in that capacity. In particular, we present indications that the two-gene module that encodes a minimal nucleotidyl transferase and the accompanying HEPN protein, and is extremely abundant in many archaea and bacteria, especially, thermophiles might comprise a novel TAS. We present a survey of previously known and newly predicted TAS in 750 complete genomes of archaea and bacteria, quantitatively demonstrate the exceptional mobility of the TAS, and explore the network of toxin-antitoxin pairings that combines plasticity with selectivity. Conclusion The defining properties of the TAS, namely, the typically small size of the toxin and antitoxin genes, fast evolution, and extensive horizontal mobility

  13. Cost-effectiveness of treating upper limb spasticity due to stroke with botulinum toxin type A: results from the botulinum toxin for the upper limb after stroke (BoTULS) trial.

    PubMed

    Shackley, Phil; Shaw, Lisa; Price, Christopher; van Wijck, Frederike; Barnes, Michael; Graham, Laura; Ford, Gary A; Steen, Nick; Rodgers, Helen

    2012-12-01

    Stroke imposes significant burdens on health services and society, and as such there is a growing need to assess the cost-effectiveness of stroke treatment to ensure maximum benefit is derived from limited resources. This study compared the cost-effectiveness of treating post-stroke upper limb spasticity with botulinum toxin type A plus an upper limb therapy programme against the therapy programme alone. Data on resource use and health outcomes were prospectively collected for 333 patients with post-stroke upper limb spasticity taking part in a randomized trial and combined to estimate the incremental cost per quality adjusted life year (QALY) gained of botulinum toxin type A plus therapy relative to therapy alone. The base case incremental cost-effectiveness ratio (ICER) of botulinum toxin type A plus therapy was £93,500 per QALY gained. The probability of botulinum toxin type A plus therapy being cost-effective at the England and Wales cost-effectiveness threshold value of £20,000 per QALY was 0.36. The point estimates of the ICER remained above £20,000 per QALY for a range of sensitivity analyses, and the probability of botulinum toxin type A plus therapy being cost-effective at the threshold value did not exceed 0.39, regardless of the assumptions made.

  14. Cost-Effectiveness of Treating Upper Limb Spasticity Due to Stroke with Botulinum Toxin Type A: Results from the Botulinum Toxin for the Upper Limb after Stroke (BoTULS) Trial

    PubMed Central

    Shackley, Phil; Shaw, Lisa; Price, Christopher; van Wijck, Frederike; Barnes, Michael; Graham, Laura; Ford, Gary A.; Steen, Nick; Rodgers, Helen

    2012-01-01

    Stroke imposes significant burdens on health services and society, and as such there is a growing need to assess the cost-effectiveness of stroke treatment to ensure maximum benefit is derived from limited resources. This study compared the cost-effectiveness of treating post-stroke upper limb spasticity with botulinum toxin type A plus an upper limb therapy programme against the therapy programme alone. Data on resource use and health outcomes were prospectively collected for 333 patients with post-stroke upper limb spasticity taking part in a randomized trial and combined to estimate the incremental cost per quality adjusted life year (QALY) gained of botulinum toxin type A plus therapy relative to therapy alone. The base case incremental cost-effectiveness ratio (ICER) of botulinum toxin type A plus therapy was £93,500 per QALY gained. The probability of botulinum toxin type A plus therapy being cost-effective at the England and Wales cost-effectiveness threshold value of £20,000 per QALY was 0.36. The point estimates of the ICER remained above £20,000 per QALY for a range of sensitivity analyses, and the probability of botulinum toxin type A plus therapy being cost-effective at the threshold value did not exceed 0.39, regardless of the assumptions made. PMID:23342679

  15. Characterization of an organism that produces type E botulinal toxin but which resembles Clostridium butyricum from the feces of an infant with type E botulism.

    PubMed Central

    McCroskey, L M; Hatheway, C L; Fenicia, L; Pasolini, B; Aureli, P

    1986-01-01

    The apparent causative organism from the only reported case of type E infant botulism was isolated and characterized. Except for its ability to produce type E botulinal toxin, this organism (strain 5262) would be unquestionably identified as Clostridium butyricum. This is the second time an organism resembling a defined Clostridium species other than a member of the C. botulinum group has been implicated in infant botulism. PMID:3517043

  16. Lower facial remodeling with botulinum toxin type A for the treatment of masseter hypertrophy*

    PubMed Central

    Klein, Fernanda Homem de Mello de Souza; Brenner, Fabiane Mulinari; Sato, Maurício Shigeru; Robert, Fernanda Manfron Batista Rosas; Helmer, Karin Adriane

    2014-01-01

    BACKGROUND Masseter hypertrophy has been treated with botulinum toxin injections because of esthetic complaints especially in Asians. OBJECTIVES The goal of the present study was to evaluate the efficacy of abobotulin toxin use in masseter hipertrophy treatment in Brazilians. METHODS Ten Brazilian female patients with masseter hypertrophy were subjected to injections of 90U of abobotulinum toxin A applied on each side respecting the safety zone stabilished in literature and were followed up for 24 weeks. RESULTS When analyzing the coefficients between measures of middle and lower third of the face obtained from standardized photographs, an increase was observed, with statistical significance at 2 weeks (p=0.005) and 12 weeks (p=0.001). The progression of lower third reduction was 3.94%, 5.26%, 11.99%, and 5.47% (2, 4, 12, and 24 weeks respectively). All patients showed improvement in bruxism after treatment. Observed adverse effects were masticatory fatigue, smile limitation, and smile asymmetry. CONCLUSION The use of abobotulinum toxin A for masseter hypertrophy is effective in Brazilians and reached its maximum effect of facial thinning at 12 weeks. Smile limitation had a higher incidence compared to that reported in the literature and may result from risorius muscle blockage caused by toxin dissemination. Despite its side effects, 80% of the patients would like to repeat the treatment. PMID:25387491

  17. Botulinum Toxin Type A for the Treatment of Primary Hyperhidrosis: A Prospective Study of 52 Patients.

    PubMed

    Martí, N; Ramón, D; Gámez, L; Reig, I; García-Pérez, M Á; Alonso, V; Jordá, E

    2010-09-01

    Primary hyperhidrosis is characterized by excessive sweating in a defined region of the body. It should not be considered a purely cosmetic problem as it has a significant impact on the social and professional relationships of affected individuals. The aim of this study was to determine the clinical profile of patients with primary hyperhidrosis and assess the results obtained with the use of botulinum toxin type A (BTX-A) in clinical practice. The study included 52 patients (39 women and 13 men) with a diagnosis of primary hyperhidrosis treated for the first time with BTX-A. All patients completed a questionnaire that included the following information: age; sex; profession; age at onset, family history, and site of hyperhidrosis; accompanying signs and symptoms, and previous treatment; time to effect of BTX-A; local or systemic side effects; and severity of hyperhidrosis before and after BTX-A treatment. Primary hyperhidrosis began during puberty in 61.5% of the patients included in the study, 75% were women, and the mean age was 29.9 years. In 36.5% of patients, first-degree relatives also had primary hyperhidrosis. Hyperhidrosis was classified as palmar in 61.5% of cases, plantar in 53.8%, and axillary in 59.6%. Other sites were affected less frequently. The most common accompanying symptoms were facial erythema (32.7%), palpitations (30.7%), muscle tension (28.8%), shivering (23%), and headache (17.3%). Treatment with BTX-A was well tolerated and there was a highly significant reduction in the severity of hyperhidrosis 2 months after performing the treatment (P<0.001). Copyright © 2009 Elsevier España, S.L. y AEDV. All rights reserved.

  18. Antinociceptive action of botulinum toxin type A in carrageenan-induced mirror pain.

    PubMed

    Drinovac Vlah, V; Bach-Rojecky, L; Lacković, Z

    2016-12-01

    "Mirror pain" or mirror-image pain (MP) is pain opposite to the side of injury. Mechanism and frequency in humans are not known. There is no consent on therapy. Here we report that unilaterally injected botulinum toxin type A (BT-A) has bilateral effect in experimental MP, thus deserves to be investigated as therapy for this condition. We examined the localization of BT-A's bilateral antinociceptive action in MP induced by 3 % carrageenan intramuscular injection in Wistar rats. BT-A was applied peripherally (5 U/kg), into ipsilateral or contralateral hind paw pad (i.pl.) and centrally (1 U/kg), at spinal (intrathecally, i.t.) or supraspinal (intracisternally, i.c.) level. Additionally, we examined the involvement of central opioid and GABAergic systems, as well as the contribution of peripheral capsaicin-sensitive neurons to BT-A's bilateral antinociceptive effect. Ipsilateral i.pl. and i.t. BT-A reduced the bilateral mechanical sensitivity to von Frey filaments, while contralateral i.pl. and i.c. treatments had no effect on either tested side. Bilateral antinociceptive effect of ipsilateral i.pl. BT-A was prevented by μ-opioid antagonist naloxonazine (1.5 μg/10 μl) and GABAA antagonist bicuculline (1 μg/10 μl) if applied at the spinal level, in contrast to supraspinal application of the same doses. Local treatment of sciatic nerve with 2 % capsaicin 5 days following BT-A i.pl. injection caused desensitization of sciatic capsaicin-sensitive fibers, but did not affect bilateral antinociceptive effect of BT-A and the presence of cleaved SNAP-25 at the spinal cord slices. Present experiments suggest segmental actions of peripheral BT-A at spinal level, which are probably not solely dependent on capsaicin-sensitive neurons.

  19. Botulinum toxin type A reduces capsaicin-evoked pain and neurogenic vasodilatation in human skin.

    PubMed

    Tugnoli, Valeria; Capone, Jay Guido; Eleopra, Roberto; Quatrale, Rocco; Sensi, Mariachiara; Gastaldo, Ernesto; Tola, Maria Rosaria; Geppetti, Pierangelo

    2007-07-01

    The effect of Botulinum Toxin type A (BoNT/A) on pain and neurogenic vasodilatation induced by application to the human skin of thermal stimuli and capsaicin was evaluated in a double blind study. A capsaicin cream (0.5 ml of a 0.075%) was applied to the skin of both forearms of eighteen subjects randomly pretreated with either BoNT/A (Botox) or 0.9% saline (NS). Capsaicin was applied to a skin area either inside (protocol A) or adjacent to the BoNT/A treated area (protocol B). Pre-treatment with BoNT/A did not affect thermal-specific and thermal-pain thresholds (by quantitative sensory testing). However, capsaicin-induced pain sensation (by a visual analogue scale), flare area (by acetate sheet) and changes in cutaneous blood flow (CBF, by laser Doppler flowmetry) were reduced when capsaicin was administered inside (protocol A) the BoNT/A treated area. In Protocol B, capsaicin-induced pain was unchanged, and capsaicin-induced flare/increase in CBF were reduced only in the area treated with BoNT/A, but not in the BoNT/A untreated area. Results indicate that (i) BoNT/A reduces capsaicin-induced pain and neurogenic vasodilatation without affecting the transmission of thermal and thermal-pain modalities; (ii) reduction in capsaicin-induced pain occurs only if capsaicin is administered into the BoNT/A pretreated area; (iii) reduction in neurogenic vasodilatation by BoNT/A does not contribute to its analgesic action. BoNT/A could be tested for the treatment of conditions characterised by neurogenic inflammation and inflammatory pain.

  20. Single event multilevel botulinum toxin type A treatment and surgery: similarities and differences.

    PubMed

    Molenaers, G; Desloovere, K; De Cat, J; Jonkers, I; De Borre, L; Pauwels, P; Nijs, J; Fabry, G; De Cock, P

    2001-11-01

    The present study attempts to provide objective evidence of two treatment options for children with cerebral palsy (CP): multilevel botulinum toxin type A (BTX-A) injections and multilevel surgery. The purpose of the study was to clarify the differences and the similarities, and common treatment principles of both treatment strategies. Objective three dimensional gait analysis data were studied retrospectively in two patient groups pre- and post-treatment (randomly selected from a group of children that were treated between 1998 and 1999). In the first group, 29 children with CP were managed with BTX-A injections according to an integrated multilevel approach (Molenaers et al., 1999a). A second group of 23 children with CP were managed by a more traditional single event multilevel surgery, also according to an integrated approach. Our aim was to evaluate the differences as well as the similarities between both patient groups, using a set of 56 parameters selected from three-dimensional gait analysis. The unifying concept between management with BTX-A injections and orthopaedic surgery was the adoption of a multilevel approach at one session. The groups demonstrated considerable differences with respect to age, pretreatment condition and amount and level of improvement after treatment. The children who received BTX-A were typically younger, and showed primary gait problems in the distal joints, whereas the children who underwent surgery demonstrated a higher frequency of gait deviations in the transverse plane and had more complications. Although the benefit of both treatments was confirmed by the present study, a difference in the amount and level of improvement was also demonstrated. In conclusion, these treatment modalities should be regarded as complementary rather than mutually exclusive treatments, with both calling for an integrated approach.

  1. Preoperative progressive pneumoperitoneum and botulinum toxin type A in patients with large incisional hernia.

    PubMed

    Bueno-Lledó, J; Torregrosa, A; Ballester, N; Carreño, O; Carbonell, F; Pastor, P G; Pamies, J; Cortés, V; Bonafé, S; Iserte, J

    2017-04-01

    Combination of preoperative progressive pneumoperitoneum (PPP) and botulinum toxin type A (BT) has not been previously reported in the management of large incisional hernia (LIH). Observational study of 45 consecutive patients with LIH between June 2010 and July 2014. The diameters of the hernia sac, the volumes of the incisional hernia (VIH) and the abdominal cavity (VAC), and the VIH/VAC ratio were measured before and after PPP and BT using abdominal CT scan data. We indicated the combination of both techniques when the volume of the incisional hernia (VIH)/volume of the abdominal cavity (VAC) ratio was >20%. The median insufflated volume of air for PPP was 8.600 ± 3.200 cc (4.500-13.250), over a period of 14.3 ± 1.3 days (13-16). BT administration time was 40.2 ± 3.3 days (37-44). We obtained an average value of reduction of 14% of the VIH/VAC ratio after PPP and BT (p < 0.05). Complications associated with PPP were 15.5%, and with surgical technique, 26.6%. No complications occurred during the BT administration. Reconstructive technique was anterior CST and primary fascial closure was achieved in all patients. Median follow-up was 40.5 ± 19 months (12-60) and we reported 2 cases of hernia recurrence (4.4%). Preoperative combination of PPP and BT is feasible and a useful tool in the surgical management of LIH, although at the cost of some specific complications.

  2. Evaluation of botulinum toxin type A effectiveness in preventing postoperative intraperitoneal adhesions

    PubMed Central

    Uysal, Erdal

    2017-01-01

    Purpose Postoperative intraperitoneal adhesions (PIAs) are one of the most important problems surgeons have to face after laparotomies. In this study, we aimed to evaluate the effectiveness of local application of botulinum toxin type A (BoNT-A) in various dosages on the prevention of intra-abdominal adhesions in rats with experimental intra-abdominal adhesions. Methods Forty Wistar Albino female rats were randomly separated into 4 groups. The 4 groups were determined as follows: Control (group 1, n = 10); Sham (group 2, n = 10); 10-µg/kg low-dose BoNT-A (group 3, n = 10) and 30-µg/kg high-dose BoNT-A (group 4, n = 10). Subserosal injuries were created on the caecum of all rats. Laparotomy was performed on the fifth day. Adhesion scores, histopathological examination, and E-cadherin expression levels were evaluated. Results General adhesion scores for groups 1 and 2 were determined to be significantly high when compared to group 4 (P < 0.001). A significant difference was also determined between groups 3 and 4 in terms of general adhesion scores (P < 0.05). In pair comparisons, a significant decrease in high-dose BoNT-A group (group 4) when compared to groups 1 and 2 in terms of neovascularization, fibroblast density, collagen deposition and inflammatory cell count was determined (P < 0.05). Conclusion A significant decrease was observed only in postoperative PIAs in the high-dose BoNT-A group between all 4 rat-groups with experimentally created postoperative PIAs. In this study, high-dose BoNT-A is determined to be an effective agent in preventing postoperative PIAs. PMID:28706891

  3. High-resolution crystal structure of HA33 of botulinum neurotoxin type B progenitor toxin complex

    PubMed Central

    Lee, Kwangkook; Lam, Kwok-Ho; Kruel, Anna Magdalena; Perry, Kay; Rummel, Andreas; Jin, Rongsheng

    2014-01-01

    Botulinum neurotoxins (BoNTs) are produced as progenitor toxin complexes (PTCs) by Clostridium botulinum. The PTCs are composed of BoNT and non-toxic neurotoxin-associated proteins (NAPs), which serve to protect and deliver BoNT through the gastrointestinal tract in food borne botulism. HA33 is a key NAP component that specifically recognizes host carbohydrates and helps enrich PTC on the intestinal lumen preceding its transport across the epithelial barriers. Here, we report the crystal structure of HA33 of type B PTC (HA33/B) in complex with lactose at 1.46 Å resolution. The structural comparisons among HA33 of serotypes A–D reveal two different HA33–glycan interaction modes. The glycan-binding pockets on HA33/A and B are more suitable to recognize galactose-containing glycans in comparison to the equivalent sites on HA33/C and D. On the contrary, HA33/C and D could potentially recognize Neu5Ac as an independent receptor, whereas HA33/A and B do not. These findings indicate that the different oral toxicity and host susceptibility observed among different BoNT serotypes could be partly determined by the serotype-specific interaction between HA33 and host carbohydrate receptors. Furthermore, we have identified a key structural water molecule that mediates the HA33/B–lactose interactions. It provides the structural basis for development of new receptor-mimicking compounds, which have enhanced binding affinity with HA33 through their water-displacing moiety. PMID:24631690

  4. Botulinum toxin type A reconstituted in lidocaine with epinephrine for facial rejuvenation: results of a participant satisfaction survey.

    PubMed

    Kim, Alex; Jung, Jeong; Pak, Alexis

    2013-07-01

    To assess the feasibility, safety, and lack of inferiority of reconstituting botulinum toxin type A (BTX-A) in 1% lidocaine hydrochloride with epinephrine 1:100,000, 181 participants were asked to complete a satisfaction survey 3 to 6 months after treatment with the reconstituted formulation for facial rejuvenation. The addition of lidocaine was believed to achieve an immediate paralyzing effect on the injected muscles, and the addition of epinephrine was hypothesized to minimize diffusion to adjacent muscles. Participants were treated in the areas of the forehead and glabella, as well as the orbicularis oculi, orbicularis oris, and procerus muscles, in varying doses (10-60 U). Fifty-eight percent (91/157) of participants reported being more satisfied with BTX-A reconstituted in 1% lidocaine with epinephrine 1:100,000, with 85.7% (78/91) of these participants reporting that the immediate results made the formulation superior; 35.7% (56/157) were indifferent and 6.4% (10/157) reported that the modified formulation did not work better. The injection of BTX-A reconstituted in 1% lidocaine with epinephrine 1:100,000 presented no increased adverse effects (AEs), no decrease in pharmacologic potency, immediate feedback to the clinician, and higher satisfaction for the participants who previously had been treated with BTX-A reconstituted in unpreserved saline. Botulinum toxin type A reconstituted in 1% lidocaine with epinephrine 1:100,000 may increase the duration and efficacy of this widely used toxin.

  5. [The use of the toxin binding inhibition test for potency testing of Clostridium novyi type B alpha toxoid vaccines].

    PubMed

    Luick, K; Gottschaldt, J; Schulze, F; Werner, E; Erler, W; Borrmann, E

    2002-04-01

    The toxin binding inhibition test (ToBI) were developed for potency testing of C. novyi type B alpha toxoid containing veterinary vaccines to replace the currently used toxin neutralisation test in mice (TNT). The antitoxin titres of rabbit sera (AN-, HV- and SP sera) were determined with ToBI using the international reference serum with known antitoxin titre. In order to show the validity of the methods, the results were compared with those of the manufacturers/regulatory authorities and correlation coefficients were calculated. The correlation coefficients were r = 0.93 (AN sera), r = 0.73 (HV sera) and r = 0.85 (SP sera). All correlations were statistically significant. The specificity of the methods could be proved using heterologous antisera. The results of the ToBI were reproducible. Thus, the ToBI offers a suitable in vitro method for the determination of the antitoxin titre of rabbit antisera as an alternative to the toxin neutralisation in mice for potency testing of vaccines containing C. novyi type B alpha toxoid.

  6. Botulinum toxin type A and B improve quality of life in patients with axillary and palmar hyperhidrosis.

    PubMed

    Rosell, Karolina; Hymnelius, Kristina; Swartling, Carl

    2013-05-01

    Hyperhidrosis is a common disorder that may have a severe impact on quality of life. The aim of this study was to investigate the clinical effect of two novel botulinum toxins, Xeomin®, a type A botulinum toxin, and Neuro-bloc®, a type B botulinum toxin, in the treatment of axillary and palmar hyperhidrosis. A total of 84 patients, 58 with axillary and 26 with palmar hyperhidrosis, were included in this open study. Axillae were injected with 107 ± 22 U Xeomin® and palms were injected with 213 ± 19 U Xeomin® and 264 ± 60 U Neurobloc® over the thenar eminences to avoid muscle weakness. At follow-up 3 weeks post-treatment, all patients treated for axillary hyperhidrosis reported satisfaction in self-ranking, evaporation decreased > 40%, and Dermatology Life Quality Index (DLQI) score improved from 12.0 to 1.7 (p < 0.05). In the palmar group 95% were satisfied, evaporation decreased > 50% and DLQI score improved from 10.3 to 1.2 (p < 0.05). Only one patient in the palmar group experienced muscle weakness. In conclusion, Xeomin® has an excellent effect on axillary hyperhidrosis and in combination with Neurobloc® on palmar hyperhidrosis. Neurobloc® may be an option for use in the treatment of palmar hyperhidrosis in order to minimize muscular side-effects.

  7. Anterior chemodenervation of levator palpebrae superioris with botulinum toxin type-A (Botox) to induce temporary ptosis for corneal protection.

    PubMed

    Naik, M N; Gangopadhyay, N; Fernandes, M; Murthy, R; Honavar, S G

    2008-09-01

    To evaluate the effectiveness of anterior chemodenervation of levator palpebrae superioris with Botulinum toxin type A (Botox) to induce temporary ptosis for corneal protection, and assess the incidence of superior rectus underaction. Prospective interventional case series. Patients with ocular surface pathology requiring temporary tarsorrhaphy underwent transcutaneous anterior chemodenervation of levator palpebrae superioris with Botox. The onset and duration of ptosis, corneal healing, and superior rectus underaction was evaluated. Ten eyes of 10 patients underwent transcutaneous anterior chemodenervation of levator muscle. Five patients had Bells palsy with exposure keratopathy, four patients had persistent epithelial defect, and one had neurotrophic ulcer. The median age at presentation was 30 years. Median dose of Botulinum toxin injection was 12.5 U (range 10-15 U). The mean palpebral fissure height of 9 mm (SD+/-2.1 mm) before injection, reduced to 2.8 mm (SD+/-1.9 mm) at 1-week post-injection. More than 50% reduction in palpebral fissure height was seen in nine out of 10 eyes (90%, 95% CI 71.4-100%) at 1 week, seven of nine eyes (77.8%, 95% CI 50.6-100%) at 2 weeks, and two of nine eyes (22.2%, 95% CI 0-49.4%) at 4 weeks, and returned to pretreatment level after mean duration of 9.2 weeks (range 5-16 weeks). Superior rectus underaction was not noted in any of the patient (95% CI 0-30%). Corneal pathology improved in all cases. Anterior chemodenervation of levator palpebrae superioris with Botulinum toxin type A (Botox) induces significant temporary ptosis and aids in corneal healing. Anterior placement of the toxin injection may avoid superior rectus underaction.

  8. Cinnamon Oil Inhibits Shiga Toxin Type 2 Phage Induction and Shiga Toxin Type 2 Production in Escherichia coli O157:H7

    PubMed Central

    Sheng, Lina; Rasco, Barbara

    2016-01-01

    ABSTRACT This study evaluated the inhibitory effect of cinnamon oil against Escherichia coli O157:H7 Shiga toxin (Stx) production and further explored the underlying mechanisms. The MIC and minimum bactericidal concentration (MBC) of cinnamon oil against E. coli O157:H7 were 0.025% and 0.05% (vol/vol), respectively. Cinnamon oil significantly reduced Stx2 production and the stx2 mRNA expression that is associated with diminished Vero cell cytotoxicity. Consistently, induction of the Stx-converting phage where the stx2 gene is located, along with the total number of phages, decreased proportionally to cinnamon oil concentration. In line with decreased Stx2 phage induction, cinnamon oil at 0.75× and 1.0× MIC eliminated RecA, a key mediator of SOS response, polynucleotide phosphorylase (PNPase), and poly(A) polymerase (PAP I), which positively regulate Stx-converting phages, contributing to reduced Stx-converting phage induction and Stx production. Furthermore, cinnamon oil at 0.75× and 1.0× MIC strongly inhibited the qseBC and luxS expression associated with decreased AI-2 production, a universal quorum sensing signaling molecule. However, the expression of oxidative stress response genes oxyR, soxR, and rpoS was increased in response to cinnamon oil at 0.25× or 0.5× MIC, which may contribute to stunted bacterial growth and reduced Stx2 phage induction and Stx2 production due to the inhibitory effect of OxyR on prophage activation. Collectively, cinnamon oil inhibits Stx2 production and Stx2 phage induction in E. coli O157:H7 in multiple ways. IMPORTANCE This study reports the inhibitory effect of cinnamon oil on Shiga toxin 2 phage induction and Shiga toxin 2 production. Subinhibitory concentrations (concentrations below the MIC) of cinnamon oil reduced Stx2 production, stx2 mRNA expression, and cytotoxicity on Vero cells. Subinhibitory concentrations of cinnamon oil also dramatically reduced both the Stx2 phage and total phage induction in E. coli O157:H7

  9. Vaccination against type F botulinum toxin using attenuated Salmonella enterica var Typhimurium strains expressing the BoNT/F H(C) fragment.

    PubMed

    Foynes, Susan; Holley, Jane L; Garmory, Helen S; Titball, Richard W; Fairweather, Neil F

    2003-03-07

    The utility of the htrA, pagC and nirB promoters to direct the expression of the carboxy-terminal (H(C)) fragment of botulinum toxin F (FH(C)) in Salmonella enterica var Typhimurium has been evaluated. Only low levels of serum antibody were induced after immunisation, and some protection against botulinum toxin type F was demonstrated after oral immunisation of mice with two doses of any of these recombinant Salmonella. Immunisation with two doses of recombinant Salmonella expressing FH(C) from the htrA promoter gave the greatest protection, against up to 10,000 mouse lethal doses of botulinum toxin type F. These results demonstrate the feasibility of an orally delivered vaccine against botulinum toxin type F.

  10. Clinical Aspects of the Use of Botulinum Toxin Type A in the Treatment of Dysfunction of the Masticatory System.

    PubMed

    Bogucki, Zdzisław A; Kownacka, Mariola

    2016-01-01

    The purpose of this work is to present a new, still experimental method of treating temporomandibular disorders (TMD) by injecting botulinum toxin Type A (TBX-A), using its effects not as a toxin but as a medication. The mechanism of TBX-A, indications and contraindications for its use, as well as possible side effects, are discussed. Temporomandibular disorders are of concern to approximately 70-80% of the population. The effect of botulinum toxin depends on blocking the release of acetylcholine from a presynaptic neuromuscular synapse and, in the autonomous system, blocking its release from post-ganglionic cholinergic neurons. In cases of long-term TMJ disorders, muscle activity increases and spastic contractions may even appear. TBX-A offers an opportunity for a normal social and family life for many patients suffering from masticatory system disorders (MSD), who have been isolated from the environment by pain. The study is based on a review of the literature and the authors' own experiences during several attempts to treat patients by this method. TBX-A is a safe medicine when the injection is performed by a well-trained physician.

  11. Comparative Adjuvant Effects of Type II Heat-Labile Enterotoxins in Combination with Two Different Candidate Ricin Toxin Vaccine Antigens.

    PubMed

    Vance, David J; Greene, Christopher J; Rong, Yinghui; Mandell, Lorrie M; Connell, Terry D; Mantis, Nicholas J

    2015-12-01

    Type II heat-labile enterotoxins (HLTs) constitute a promising set of adjuvants that have been shown to enhance humoral and cellular immune responses when coadministered with an array of different proteins, including several pathogen-associated antigens. However, the adjuvant activities of the four best-studied HLTs, LT-IIa, LT-IIb, LT-IIb(T13I), and LT-IIc, have never been compared side by side. We therefore conducted immunization studies in which LT-IIa, LT-IIb, LT-IIb(T13I), and LT-IIc were coadministered by the intradermal route to mice with two clinically relevant protein subunit vaccine antigens derived from the enzymatic A subunit (RTA) of ricin toxin, RiVax and RVEc. The HLTs were tested with low and high doses of antigen and were assessed for their abilities to stimulate antigen-specific serum IgG titers, ricin toxin-neutralizing activity (TNA), and protective immunity. We found that all four HLTs tested were effective adjuvants when coadministered with RiVax or RVEc. LT-IIa was of particular interest because as little as 0.03 μg when coadministered with RiVax or RVEc proved effective at augmenting ricin toxin-specific serum antibody titers with nominal evidence of local inflammation. Collectively, these results justify the need for further studies into the mechanism(s) underlying LT-IIa adjuvant activity, with the long-term goal of evaluating LT-IIa's activity in humans.

  12. Persistence of Clostridium botulinum type C toxin in blow fly (Calliphoridae) larvae as a possible cause of avian botulism in spring.

    PubMed

    Hubálek, Z; Halouzka, J

    1991-01-01

    Diverse samples were examined at a site of water-bird mortality, caused by Clostridium botulinum type C toxin in southern Moravia (Czechoslovakia). The toxin was detected in high concentrations in mute swan (Cygnus olor) carcasses (less than or equal to 1 x 10(6) LD50/g) as well as in necrophagous larvae and pupae of the blow flies Lucilia sericata and Calliphora vomitoria (less than or equal to 1 x 10(5) LD50/g) collected from them. It was detected in lower concentrations (less than or equal to 1 x 10(3) LD50/g) in other invertebrates (ptychopterid fly larvae, leeches, sow-bugs) associated with these carcasses, and occasionally in water samples (8 LD50/ml) close to the carrion. The toxin was not detected in the samples of water, mud or invertebrates collected at a distance greater than or equal to 5 m from the carcasses. The toxin-bearing larvae of L. sericata and C. vomitoria, containing 80,000 LD50/g of type C toxin, were exposed in the mud at the study site for 131 days from November to March. Although the toxin activity decreased 25-fold and 40-fold in the two samples of maggots exposed during this period, it remained very high (less than or equal to 3,200 LD50/g). Birds ingesting a relatively low number of these toxic larvae (or pupae) in the spring could receive a lethal dose of the toxin.

  13. Paralysis of the orbicularis muscle of the eye using botulinum toxin type A in the treatment for dry eye.

    PubMed

    Serna-Ojeda, Juan Carlos; Nava-Castaneda, Angel

    2017-03-01

    To evaluate the efficacy of botulinum toxin type A injection to cause orbicularis eyelid muscle paralysis to improve dry eye signs and symptoms. A prospective, randomized, comparative eye-to-eye and interventional study was performed. Patients with dry eye symptoms and positive fluorescein corneal staining were included. Randomly one eyelid received a subcutaneous injection of botulinum toxin in the medial orbicularis muscle portion of the lower eyelid, and the other eye received placebo. The subjective evaluation was achieved with a questionnaire assessing symptoms, quality of vision and ocular comfort level. The objective evaluation included the measurement of the tear film break-up time (TBUT), Schirmer's test and corneal and conjunctival staining. Twenty patients were included with a mean age of 59.5 years. Two weeks after the botulinum toxin injection, all patients showed a decrease in the horizontal movement of the lower eyelid when blinking. The eyes in the active treatment group showed better scores compared with the sham group in four symptoms 4 weeks after the treatment. The TBUT was higher at 1 and 3 months in the active treatment group. The corneal and conjunctival staining were significantly lower in the active treatment group at 1 and 3 months, and the Schirmer's test showed better measurements in the same group at 2 weeks, 1 month and 3 months. There were no adverse events reported. The injection of botulinum toxin A in the medial part of the lower eyelid is an effective and safe procedure that temporally improves some of the signs and symptoms of patients with dry eye. © 2016 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  14. Botulinum toxin type A in post-stroke upper limb spasticity.

    PubMed

    Kaji, Ryuji; Osako, Yuka; Suyama, Kazuaki; Maeda, Toshio; Uechi, Yasuyuki; Iwasaki, Masaru

    2010-08-01

    To evaluate the efficacy and safety of one-time injections of botulinum toxin type A (BoNTA) in Japanese patients with post-stroke upper limb spasticity. In a multicentre, randomised, double-blind, parallel-group, placebo-controlled study, 109 patients with upper limb spasticity were randomised to receive a single treatment with lower-dose (120-150 U) or higher-dose (200-240 U) BoNTA or placebo into upper limb muscles. NCT00460564. The tone of the wrist flexor was assessed at baseline and at weeks 0, 1, 4, 6, 8 and 12 using the Modified Ashworth Scale (MAS) for wrist, finger, thumb and disability in activities of daily living (ADL) was rated using the 4-point Disability Assessment Scale (DAS). The primary endpoint was area under the curve (AUC) of the change from baseline in the MAS wrist score in the higher-dose groups. Subjects were randomised with 51 in the higher BoNTA group, 26 in the higher-dose placebo group, 21 in the lower BoNTA group and 11 in the lower-dose placebo group. Significant improvement in spasticity with higher-dose BoNTA was demonstrated by a mean difference in the AUC of the change from baseline in the MAS wrist score between the higher-dose BoNTA group and the higher-dose placebo group of -6.830 (p < 0.001, t-test), no significant different was demonstrated between the lower-dose BoNTA group and the lower-dose placebo group (p = 0.215, t-test). Significant improvements with higher-dose BoNTA were also observed in the DAS scores for limb position (p = 0.001-0.022) at all time points and dressing (p = 0.018-0.038, Wilcoxon test) at weeks 6, 8 and 12. No clinically relevant difference was noted in the frequency of treatment-related adverse events between BoNTA-treated and placebo-treated patients. The long-term efficacy and safety, and the effects on rehabilitation of BoNTA on upper limb will be evaluated using the data obtained in the open-label phase. Higher-dose BoNTA reduced spasticity in upper limb muscles and improved ADL performance in

  15. Effect of botulinum toxin type-A in patients with focal spasticity

    PubMed Central

    Selimoglu, Esra; Turgut, Selin Turan; Akpinar, Pinar; Yumusakhuylu, Yasemin; Haliloglu, Sema; Baklacioglu, Hatice Sule; Icagasioglu, Afitap

    2015-01-01

    OBJECTIVE: To investigate the effect of botulinum toxin type-A (BTX-A) on spasticity and function in patients with focal spasticity. METHODS: Patients attended to the outpatient clinic of physical medicine and rehabilitation department with a diagnosis of focal spasticity and had BTX-A injections because of spasticty were evaluated for the study. Demographic data, exercise status, orthoses, drugs used for spasticity, functional status, stages of spasticity of muscles before and after 1st and 3rd months of BTX-A injection according to Modified Ashworth Scale (MAS) were evaluated retrospectively. MedCalc 11.6 statistical program was used for statistical analyses. Statistical significance was defined as p<0.05. RESULTS: Forty-nine patients with focal spasticity were recruited for the study (35 men, 14 women). Mean age of the patients was 21.59±20.09 years. The patients had cerebral palsy (CP, n=28), 19 had hemiplegia (n=19) and paraplegia (n=2). Forty-three patients were using orthoses and exercising regularly. Mean Pediatric Functional Independence Measurement (WeeFIM) scores of the patients with CP was 54.82±28.91 and according to the Gross Motor Function Classification System (GMFCS) the patients were in stages 2 (14%), 3 (46%), 4 (11%) and 5 (29%). Mean Functional Independence Measure (FIM) of hemiplegic and paraplegic patients was 80.80±20.88. Brunnstrom staging scores for upper extremity (3.52±0.96), hands (2.68±0.82), lower extremity (4.57±1.01) were calculated. MAS muscles demonstrated statistically significant decrease in spasticity at the end of first and third months (p<0.05). CONCLUSION: We saw a significant decrease in the spasticity of upper and lower extremities in patients with focal spasiticity who received BTX-A injections. We suggest that if BTX-A injections are supported with orthoses and exercise programs, then functional status of the patients would be better. PMID:28058322

  16. C-type natriuretic peptide modulates quorum sensing molecule and toxin production in Pseudomonas aeruginosa.

    PubMed

    Blier, Anne-Sophie; Veron, Wilfried; Bazire, Alexis; Gerault, Eloïse; Taupin, Laure; Vieillard, Julien; Rehel, Karine; Dufour, Alain; Le Derf, Franck; Orange, Nicole; Hulen, Christian; Feuilloley, Marc G J; Lesouhaitier, Olivier

    2011-07-01

    Pseudomonas aeruginosa coordinates its virulence expression and establishment in the host in response to modification of its environment. During the infectious process, bacteria are exposed to and can detect eukaryotic products including hormones. It has been shown that P. aeruginosa is sensitive to natriuretic peptides, a family of eukaryotic hormones, through a cyclic nucleotide-dependent sensor system that modulates its cytotoxicity. We observed that pre-treatment of P. aeruginosa PAO1 with C-type natriuretic peptide (CNP) increases the capacity of the bacteria to kill Caenorhabditis elegans through diffusive toxin production. In contrast, brain natriuretic peptide (BNP) did not affect the capacity of the bacteria to kill C. elegans. The bacterial production of hydrogen cyanide (HCN) was enhanced by both BNP and CNP whereas the production of phenazine pyocyanin was strongly inhibited by CNP. The amount of 2-heptyl-4-quinolone (HHQ), a precursor to 2-heptyl-3-hydroxyl-4-quinolone (Pseudomonas quinolone signal; PQS), decreased after CNP treatment. The quantity of 2-nonyl-4-quinolone (HNQ), another quinolone which is synthesized from HHQ, was also reduced after CNP treatment. Conversely, both BNP and CNP significantly enhanced bacterial production of acylhomoserine lactone (AHL) [e.g. 3-oxo-dodecanoyl-homoserine lactone (3OC12-HSL) and butanoylhomoserine lactone (C4-HSL)]. These results correlate with an induction of lasI transcription 1 h after bacterial exposure to BNP or CNP. Concurrently, pre-treatment of P. aeruginosa PAO1 with either BNP or CNP enhanced PAO1 exotoxin A production, via a higher toxA mRNA level. At the same time, CNP led to elevated amounts of algC mRNA, indicating that algC is involved in C. elegans killing. Finally, we observed that in PAO1, Vfr protein is essential to the pro-virulent effect of CNP whereas the regulator PtxR supports only a part of the CNP pro-virulent activity. Taken together, these data reinforce the hypothesis that during

  17. Effect of Botulinum Toxin Type A on a Rat Surgical Wound Model

    PubMed Central

    Lee, Byung-Joo; Jeong, Jae-Hwi; Wang, Soo-Geun; Lee, Jin-Choon; Kim, Hwal-Woong

    2009-01-01

    Objectives The tension on a wound is one of the important factors that determine the degree of fibrosis and scar formation. We hypothesized that local botulinum toxin type A (Botox) induced paralysis of the musculature subjacent to a surgical wound with a skin defect would minimize the repetitive tensile forces on the surgical wound's edges, and this will result in a decreased fibroplastic response and fibrosis of the wound. Methods This is a prospective randomized experimental study. Two distinct surgical wounds were made to the dorsum of 15 adult rats, respectively. One of the 2 wounds was injected with Botox, and the other wound was used as a control, and this was done for all the rats' wounds. We evaluated the wound size, the degree of fibrosis and inflammation, the blood vessel proliferation, the thickness of the wound and the expression of transforming growth factor (TGF)-β1 in the wounds. Results There were significant differences of wound size at the 3rd and 4th week between the Botox and control groups (P<0.05). The Botox group showed less infiltration of inflammatory cells than the control group at the 2nd week (P<0.05). The Botox group showed a smaller number of fibroblasts and less fibrosis than the control group at the 4th week (P<0.05). The Botox group showed much strong collagen density than the control group at the 8th week (P<0.05). For the immunohistochemical staining, there was a lower transforming growth factor (TGF)-β1 expression in the Botox group than that of the control group at the 4th week (P<0.05). Conclusion The wounds of the Botox-treated group showed a larger wound size, less infiltration of inflammatory cells and less fibrosis, a much greater amount of collagen and a lower expression of TGF-β1 than did the control group. Botox might be used to decrease the fibrosis of a surgical wound without damaging the epithelial growth in situations for which decreased fibrosis is necessary, such as for treating laryngeal, tracheal and nasal

  18. Quantification of toxin-encoding mRNA from Clostridium botulinum type E in media containing sorbic acid or sodium nitrite by competitive RT-PCR.

    PubMed

    Sharkey, Freddie H; Markos, Spiros I; Haylock, Richard W

    2004-03-19

    Competitive reverse transcription polymerase chain reaction (cRT-PCR) was used to quantify the toxin-encoding mRNA production of a Clostridium botulinum type E strain in media containing either sorbic acid or sodium nitrite. A 10-fold reduction in toxin mRNA production and a 25-fold reduction in the proportion of toxin mRNA to total RNA, was estimated when either 1 mg ml(-1) sorbic acid or 100 microg ml(-1) sodium nitrite were added to the medium at pH 7.0.

  19. Botulinum Toxin

    DTIC Science & Technology

    2009-01-01

    1980) and the other in gibbons (Hylobates lar) (Smith et aI., 1985). In addition to nonhuman primates , most other animal species that show some...nonhuman primate species are known to be susceptible to type Cl and D toxins both in nature and as experimental models. A large natural outbreak of... primates were previously reported, one in squirrel monkeys (Saimiri sciureus) and capuchin monkeys (Cebus capucinus and Cebus olivaceus) (Smart et al

  20. Capturing the interaction types of two Bt toxins Cry1Ac and Cry2Ab on suppressing the cotton bollworm by using multi-exponential equations.

    PubMed

    Shi, Pei-Jian; Wei, Ji-Zhen; Sandhu, Hardev S; Liang, Ge-Mei

    2016-08-01

    Transgenic crops are increasingly promoted for their practical effects on suppressing certain insect pests, but all transgenic crops are not equally successful. The insect pests can easily develop resistance against single Bacillus thuringiensis (Bt) toxin transgenic crops. Therefore, transgenic crops including two or more mixed Bt-toxins can solve this problem by delaying the resistance development and killing the majority of targeted pests before the evolution of resistance. It is important to test the controlling effects of transgenic crops including multiple mixed toxins on a particular insect pest. Previous research has checked the cross-resistance and interactions between Bt toxins Cry1Ac and Cry2Ab against one susceptible and four resistant strains of cotton bollworm. The results showed that independence was the main interaction type between two toxins for the susceptible strain, whereas synergism was the main interaction type for any one resistant strain. However, the optimal combinations of two toxins were not obtained. In the present study, we developed two multi-exponential equations (namely bi- and tri-exponential equations) to describe the combination effects of two Bt toxins. Importantly, the equations can provide predictions of combination effects of different continuous concentrations of two toxins. We compared these two multi-exponential equations with the generalized linear model (GLM) in describing the combination effects, and found that the bi- and tri-exponential equations are better than GLM. Moreover, the bi-exponential equation can also provide the optimal dose combinations for two toxins. © 2015 Institute of Zoology, Chinese Academy of Sciences.

  1. Comparison of extracorporeal shock wave therapy with botulinum toxin type A in the treatment of plantar fasciitis.

    PubMed

    Roca, Bernardino; Mendoza, María A; Roca, Manuel

    2016-10-01

    To compare the efficacy of extracorporeal shock wave therapy (ESWT) with botulinum toxin type A (BoNT-A) in the treatment of plantar fasciitis (PF). Open label, prospective, randomized study. A total of 72 patients were included. In all participants the median (and interquartile range) of the visual analog scale (VAS) of pain result, when taking the first steps, was 8 (6-9) points before treatment and 6 (4-8) points after treatment (p < 0.001). In the group of patients that received ESWT, the median (and interquartile range) of improvement in the VAS of pain result, when taking the first steps, was 2 (1-4) points, and in the group of patients that received BoNT-A the same result was 1 (0-2) points (p = 0.009). In the group of patients that received ESWT, the median (and interquartile range) of improvement in the Roles and Maudsley scale of pain result was 1 (0-1) points, and in the group of patients that received BoNT-A the same result was 0 (0-1) points (p = 0.006). In a multivariate analysis use of ESWT and lower weight were associated with improvement of pain with treatment in at least one of the three VAS of pain scales used in the study. ESWT was superior to BoNT-A in the control of pain in patients with PF. Implications for Rehabilitation Plantar fasciitis is characterized by pain at the calcaneal origin of the plantar fascia, exacerbated by weight bearing after prolonged periods of rest. Although studies comparing extracorporeal shock wave therapy or botulinum toxin type A to placebo suggest a superiority of the first one, no reliable data exist about it. Extracorporeal shock wave therapy was superior to botulinum toxin type A in the control of pain in patients with PF.

  2. Exfoliated black phosphorus gas sensing properties at room temperature

    NASA Astrophysics Data System (ADS)

    Donarelli, M.; Ottaviano, L.; Giancaterini, L.; Fioravanti, G.; Perrozzi, F.; Cantalini, C.

    2016-06-01

    Room temperature gas sensing properties of chemically exfoliated black phosphorus (BP) to oxidizing (NO2, CO2) and reducing (NH3, H2, CO) gases in a dry air carrier have been reported. To study the gas sensing properties of BP, chemically exfoliated BP flakes have been drop casted on Si3N4 substrates provided with Pt comb-type interdigitated electrodes in N2 atmosphere. Scanning electron microscopy and x-ray photoelectron spectroscopy characterizations show respectively the occurrence of a mixed structure, composed of BP coarse aggregates dispersed on BP exfoliated few layer flakes bridging the electrodes, and a clear 2p doublet belonging to BP, which excludes the occurrence of surface oxidation. Room temperature electrical tests in dry air show a p-type response of multilayer BP with measured detection limits of 20 ppb and 10 ppm to NO2 and NH3 respectively. No response to CO and CO2 has been detected, while a slight but steady sensitivity to H2 has been recorded. The reported results confirm, on an experimental basis, what was previously theoretically predicted, demonstrating the promising sensing properties of exfoliated BP.

  3. Preoperative preparation of «loss of domain» hernia. Progressive pneumoperitoneum and botulinum toxin type A.

    PubMed

    Bueno-Lledó, José; Torregrosa Gallud, Antonio; Jiménez Rosellón, Raquel; Carbonell Tatay, Fernando; García Pastor, Providencia; Bonafé Diana, Santiago; Iserte Hernández, José

    2017-05-01

    Preoperative progressive pneumoperitoneum and botulinum toxin type A are useful tools in the preparation of patients with loss of domain hernias. Both procedures are complementary in the surgical repair, especially with the use of prosthetic techniques without tension, that allow a integral management of these patients. The aim of this paper is to update concepts related to both procedures, emphasizing the advantages that take place in the preoperative management of loss of domain hernias. Copyright © 2017 AEC. Publicado por Elsevier España, S.L.U. All rights reserved.

  4. Role of motor end plate-targeted Botulinum toxin type A injections in children with cerebral palsyitle.

    PubMed

    Van Campenhout, Anja; Bar-On, Lynn; Desloovere, Kaat; Molenaers, Guy

    2015-06-01

    Botulinum toxin type A (BTX) injections are frequently used in children with cerebral palsy (CP) to control spasticity. Injection variables still lead to variable outcomes of this treatment. Using instrumented spasticity assessment and muscle volume assessment the most effective location of the injection was demonstrated for gracilis and psoas muscles in children with CP. It was found that this treatment is most effective when injected in the motor endplate zones of the selected muscles. This review article presents all available research on the role of motor endplate-targeting of BTX injections in children with CP.

  5. Intrapouch injections of botulinum toxin type A for the management of unit contractions of a continent urinary diversion

    PubMed Central

    Gharajeh, Arash; Steele, Stephen S.; Siemens, D. Robert

    2008-01-01

    A 43-year-old woman presented with abdominal pain associated with incontinence from her Indiana pouch continent urinary diversion due to significant unit contractions. The patient’s symptoms were refractory to conservative management, including oral and intrapouch antimuscarinic agents. Prior to surgical reconstruction, a trial of intrapouch injections of botulinum toxin type A (BT-A) significantly improved both the abdominal pain and the incontinence. The benefit lasted about 6 months and was subsequently repeated for recurrent symptoms. To our knowledge, this is the first reported case of the management of complications of a continent urinary diversion with BT-A injections. PMID:18781211

  6. Toward Single Atom Chains with Exfoliated Tellurium.

    PubMed

    Churchill, Hugh O H; Salamo, Gregory J; Yu, Shui-Qing; Hironaka, Takayuki; Hu, Xian; Stacy, Jeb; Shih, Ishiang

    2017-08-10

    We demonstrate that the atom chain structure of Te allows it to be exfoliated as ultra-thin flakes and nanowires. Atomic force microscopy of exfoliated Te shows that thicknesses of 1-2 nm and widths below 100 nm can be exfoliated with this method. The Raman modes of exfoliated Te match those of bulk Te, with a slight shift (4 cm(-1)) due to a hardening of the A1 and E modes. Polarized Raman spectroscopy is used to determine the crystal orientation of exfoliated Te flakes. These experiments establish exfoliation as a route to achieve nanoscale trigonal Te while also demonstrating the potential for fabrication of single atom chains of Te.

  7. Granite Exfoliation, Cosumnes River Watershed, Somerset, California

    NASA Astrophysics Data System (ADS)

    Crockett, I. Q.; Neiss-Cortez, M.

    2015-12-01

    In the Sierra Nevada foothills of California there are many exposed granite plutons within the greater Sierra Nevada batholith. As with most exposed parts of the batholith, these granite slabs exfoliate. It is important to understand exfoliation for issues of public safety as it can cause rock slides near homes, roads, and recreation areas. Through observation, measuring, and mapping we characterize exfoliation in our Cosumnes River watershed community.

  8. Efficacy and Safety of Botulinum Toxin Type A in the Treatment of Glabellar Lines: A Meta-Analysis of Randomized, Placebo-Controlled, Double-Blind Trials.

    PubMed

    Guo, Yu; Lu, Yongzhou; Liu, Tianyi; Zhou, Yiqun; Yang, Ping; Zhu, Jingjing; Chen, Liang; Yang, Qingjian

    2015-09-01

    The uses of botulinum toxin type A for facial aesthetic procedures have been reported in recently published studies. The authors systematically analyzed the prospective, randomized, controlled trials, which continue to expand. New efficacy data and endpoints regarding the safety of botulinum toxin type A injection for treating glabellar lines were analyzed. The authors identified randomized controlled trials of botulinum toxin type A through searches of Ovid MEDLINE, PubMed, Elsevier, and the Cochrane Library from January of 2002 to November of 2014. The search terms included "botulinum toxin" and "glabellar lines." Only randomized, placebo-controlled, double-blind trials that used an injection dose of 20 units were included in the analysis. Safety was assessed by means of a meta-analysis of the number and frequency of adverse events. Seven studies involving 1474 subjects met inclusion criteria and qualified for meta-analysis. Overall, the pooled effective rate assessed by investigators in botulinum toxin type A treatment groups was significantly higher than that in controls (relative risk, 33.54; 95 percent CI, 18.65 to 60.33). The effective rate in treatment groups using a new endpoint was also higher than that in controls (relative risk, 99.04; 95 percent CI, 14.0 to 700.58). Subgroup analysis confirmed that botulinum toxin type A could improve the appearance of glabellar lines at rest (relative risk, 5.88; 95 percent CI, 3.49 to 9.91). There were no significant differences in the frequency of adverse events between the treatment and placebo groups in any of the studies. This meta-analysis shows that a single 20-unit dose of botulinum toxin type A is considered remarkably effective and safe for the treatment of glabellar lines. Therapeutic, II.

  9. Botulinum toxin type-A in the prophylactic treatment of medication-overuse headache: a multicenter, double-blind, randomized, placebo-controlled, parallel group study.

    PubMed

    Sandrini, Giorgio; Perrotta, Armando; Tassorelli, Cristina; Torelli, Paola; Brighina, Filippo; Sances, Grazia; Nappi, Giuseppe

    2011-08-01

    Medication-overuse headache (MOH) represents a severely disabling condition, with a low response to prophylactic treatments. Recently, consistent evidences have emerged in favor of botulinum toxin type-A (onabotulinum toxin A) as prophylactic treatment in chronic migraine. In a 12-week double-blind, parallel group, placebo-controlled study, we tested the efficacy and safety of onabotulinum toxin A as prophylactic treatment for MOH. A total of 68 patients were randomized (1:1) to onabotulinum toxin A (n = 33) or placebo (n = 35) treatment and received 16 intramuscular injections. The primary efficacy end point was mean change from baseline in the frequency of headache days for the 28-day period ending with week 12. No significant differences between onabotulinum toxin A and placebo treatment were detected in the primary (headache days) end point (12.0 vs. 15.9; p = 0.81). A significant reduction was recorded in the secondary end point, mean acute pain drug consumption at 12 weeks in onabotulinum toxin A-treated patients when compared with those with placebo (12.1 vs. 18.0; p = 0.03). When we considered the subgroup of patients with pericranial muscle tenderness, we recorded a significant improvement in those treated with onabotulinum toxin A compared to placebo treated in both primary (headache days) and secondary end points (acute pain drug consumption, days with drug consumption), as well as in pain intensity and disability measures (HIT-6 and MIDAS) at 12 weeks. Onabotulinum toxin A was safe and well tolerated, with few treatment-related adverse events. Few subjects discontinued due to adverse events. Our data identified the presence of pericranial muscle tenderness as predictor of response to onabotulinum toxin A in patients with complicated form of migraine such as MOH, the presence of pericranial muscle tenderness and support it as prophylactic treatment in these patients.

  10. Exfoliative cytology for diagnosing oral cancer.

    PubMed

    Pérez-Sayánsm, M; Somoza-Martín, J M; Barros-Angueira, F; Reboiras-López, M D; Gándara-Vila, P; Gándara Rey, J M; García-García, A

    2010-04-28

    Exfoliative cytology is a minimally invasive technique for obtaining oral cell specimens from patients for diagnostic purposes. Classical applications of oral cytology studies, such as oral candidiasis, have been extended to include oral precancerous and cancerous lesions. A number of analytical methods are available for studying cytology specimens. The development of molecular analysis techniques, the oral cancer etiopathogenic process, and improvements in liquid-based exfoliative cytology are leading to renewed interest in exfoliative cytology. Results sometimes are disputed, so the aim of our review was to clarify the applicability of exfoliative cytology to the diagnosis of oral precancerous and cancerous lesions.

  11. Global Aesthetics Consensus: Botulinum Toxin Type A--Evidence-Based Review, Emerging Concepts, and Consensus Recommendations for Aesthetic Use, Including Updates on Complications.

    PubMed

    Sundaram, Hema; Signorini, Massimo; Liew, Steven; Trindade de Almeida, Ada R; Wu, Yan; Vieira Braz, André; Fagien, Steven; Goodman, Greg J; Monheit, Gary; Raspaldo, Hervé

    2016-03-01

    Botulinum toxin type A injection remains the leading nonsurgical cosmetic procedure worldwide, with a high rate of efficacy and patient satisfaction. A multinational, multidisciplinary group of plastic surgeons and dermatologists convened the Global Aesthetics Consensus Group to develop updated consensus recommendations with a worldwide perspective for botulinum toxin and hyaluronic acid fillers. This publication on botulinum toxin type A considers advances in facial analysis, injection techniques, and avoidance and management of complications. Use of botulinum toxin has evolved from the upper face to also encompass the lower face, neck, and midface. The Global Aesthetics Consensus Group emphasizes an integrative, diagnostic approach. Injection dosage and placement are based on analysis of target muscles in the context of adjacent ones and associated soft and hard tissues. The indication for selection of botulinum toxin as a primary intervention is that excessive muscular contraction is the primary etiology of the facial disharmony to be addressed. Global Aesthetics Consensus Group recommendations demonstrate a paradigm shift toward neuromodulation rather than paralysis, including lower dosing of the upper face, more frequent combination treatment with hyaluronic acid fillers, and intracutaneous injection where indicated to limit depth and degree of action. The accumulation of clinical evidence and experience with botulinum toxin has led to refinements in treatment planning and implementation. The Global Aesthetics Consensus Group advocates an etiology-driven, patient-tailored approach, to enable achievement of optimal efficacy and safety in patient populations that are rapidly diversifying with respect to ethnicity, gender, and age. Therapeutic, V.

  12. Spatial, Temporal, and Matrix Variability of Clostridium botulinum Type E Toxin Gene Distribution at Great Lakes Beaches.

    PubMed

    Wijesinghe, Rasanthi U; Oster, Ryan J; Haack, Sheridan K; Fogarty, Lisa R; Tucker, Taaja R; Riley, Stephen C

    2015-07-01

    Clostridium botulinum type E toxin is responsible for extensive mortality of birds and fish in the Great Lakes. The C. botulinum bontE gene that produces the type E toxin was amplified with quantitative PCR from 150 sloughed algal samples (primarily Cladophora species) collected during summer 2012 from 10 Great Lakes beaches in five states; concurrently, 74 sediment and 37 water samples from four sites were also analyzed. The bontE gene concentration in algae was significantly higher than in water and sediment (P < 0.05), suggesting that algal mats provide a better microenvironment for C. botulinum. The bontE gene was detected most frequently in algae at Jeorse Park and Portage Lake Front beaches (Lake Michigan) and Bay City State Recreation Area beach on Saginaw Bay (Lake Huron), where 77, 100, and 83% of these algal samples contained the bontE gene, respectively. The highest concentration of bontE was detected at Bay City (1.98 × 10(5) gene copies/ml of algae or 5.21 × 10(6) g [dry weight]). This study revealed that the bontE gene is abundant in the Great Lakes but that it has spatial, temporal, and matrix variability. Further, embayed beaches, low wave height, low wind velocity, and greater average water temperature enhance the bontE occurrence. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  13. Biochemical and Electrophysiological Characterization of Two Sea Anemone Type 1 Potassium Toxins from a Geographically Distant Population of Bunodosoma caissarum

    PubMed Central

    Orts, Diego J. B.; Peigneur, Steve; Madio, Bruno; Cassoli, Juliana S.; Montandon, Gabriela G.; Pimenta, Adriano M. C.; Bicudo, José E. P. W.; Freitas, José C.; Zaharenko, André J.; Tytgat, Jan

    2013-01-01

    Sea anemone (Cnidaria, Anthozoa) venom is an important source of bioactive compounds used as tools to study the pharmacology and structure-function of voltage-gated K+ channels (KV). These neurotoxins can be divided into four different types, according to their structure and mode of action. In this work, for the first time, two toxins were purified from the venom of Bunodosoma caissarum population from Saint Peter and Saint Paul Archipelago, Brazil. Sequence alignment and phylogenetic analysis reveals that BcsTx1 and BcsTx2 are the newest members of the sea anemone type 1 potassium channel toxins. Their functional characterization was performed by means of a wide electrophysiological screening on 12 different subtypes of KV channels (KV1.1–KV1.6; KV2.1; KV3.1; KV4.2; KV4.3; hERG and Shaker IR). BcsTx1 shows a high affinity for rKv1.2 over rKv1.6, hKv1.3, Shaker IR and rKv1.1, while Bcstx2 potently blocked rKv1.6 over hKv1.3, rKv1.1, Shaker IR and rKv1.2. Furthermore, we also report for the first time a venom composition and biological activity comparison between two geographically distant populations of sea anemones. PMID:23466933

  14. Spatial, Temporal, and Matrix Variability of Clostridium botulinum Type E Toxin Gene Distribution at Great Lakes Beaches

    PubMed Central

    Oster, Ryan J.; Haack, Sheridan K.; Fogarty, Lisa R.; Tucker, Taaja R.; Riley, Stephen C.

    2015-01-01

    Clostridium botulinum type E toxin is responsible for extensive mortality of birds and fish in the Great Lakes. The C. botulinum bontE gene that produces the type E toxin was amplified with quantitative PCR from 150 sloughed algal samples (primarily Cladophora species) collected during summer 2012 from 10 Great Lakes beaches in five states; concurrently, 74 sediment and 37 water samples from four sites were also analyzed. The bontE gene concentration in algae was significantly higher than in water and sediment (P < 0.05), suggesting that algal mats provide a better microenvironment for C. botulinum. The bontE gene was detected most frequently in algae at Jeorse Park and Portage Lake Front beaches (Lake Michigan) and Bay City State Recreation Area beach on Saginaw Bay (Lake Huron), where 77, 100, and 83% of these algal samples contained the bontE gene, respectively. The highest concentration of bontE was detected at Bay City (1.98 × 105 gene copies/ml of algae or 5.21 × 106 g [dry weight]). This study revealed that the bontE gene is abundant in the Great Lakes but that it has spatial, temporal, and matrix variability. Further, embayed beaches, low wave height, low wind velocity, and greater average water temperature enhance the bontE occurrence. PMID:25888178

  15. What Is the Link between Stringent Response, Endoribonuclease Encoding Type II Toxin-Antitoxin Systems and Persistence?

    PubMed

    Ramisetty, Bhaskar C M; Ghosh, Dimpy; Roy Chowdhury, Maoumita; Santhosh, Ramachandran S

    2016-01-01

    Persistence is a transient and non-inheritable tolerance to antibiotics by a small fraction of a bacterial population. One of the proposed determinants of bacterial persistence is toxin-antitoxin systems (TASs) which are also implicated in a wide range of stress-related phenomena. Maisonneuve E, Castro-Camargo M, Gerdes K. 2013. Cell 154:1140-1150 reported an interesting link between ppGpp mediated stringent response, TAS, and persistence. It is proposed that accumulation of ppGpp enhances the accumulation of inorganic polyphosphate which modulates Lon protease to degrade antitoxins. The decrease in the concentration of antitoxins supposedly activated the toxin to increase in the number of persisters during antibiotic treatment. In this study, we show that inorganic polyphosphate is not required for transcriptional activation of yefM/yoeB TAS, which is an indirect indication of Lon-dependent degradation of YefM antitoxin. The Δ10 strain, an Escherichia coli MG1655 derivative in which the 10 TAS are deleted, is more sensitive to ciprofloxacin compared to wild type MG1655. Furthermore, we show that the Δ10 strain has relatively lower fitness compared to the wild type and hence, we argue that the persistence related implications based on Δ10 strain are void. We conclude that the transcriptional regulation and endoribonuclease activity of YefM/YoeB TAS is independent of ppGpp and inorganic polyphosphate. Therefore, we urge for thorough inspection and debate on the link between chromosomal endoribonuclease TAS and persistence.

  16. 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

    PubMed Central

    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. tObjective: The aim of this study was to examine the severity, duration, and impact of xerostomia in patients with cervical dystonia who reported this AE in routine clinical practice following treatment with botulinum toxin type B. Methods: In this uncontrolled study, investigators at 5 study centers across the United States retrospectively identified patients who were diagnosed with cervical dystonia and had received ≥ 1 treatment with botulinum toxin type B injection and who had reported xerostomia, based on patients' charts. These patients were mailed a survey that included questions about their treatment history, disease severity, and xerostomia (severity, onset, duration, change with subsequent injections, and effects on dental and oral health), as well as an 8-item Patient Benefit Questionnaire (PBQ), which was designed to assess the impact of xerostomia symptoms on patients' ADLs. Results: A total of 45 patients received a mean of 2.91 injections with botulinum toxin type B (mean dose per injection, 11,958 U), with a total of 131 injections. The mean severity of patient-rated xerostomia following the first injection of botulinum toxin type B was 3.88 on a scale of 1 (mild) to 5 (severe), and this rating did not change for patients who received subsequent injections (mean, 3.76). Following atypical injection of botulinum toxin type B, xerostomia began a mean (SD) of 4.82 (3.32) days later and persisted for a mean (SD) duration of 5.56 (3.57) weeks. The overall mean score on the 10-point PBQ prior to botulinum toxin treatment was 8.89, which decreased to 5.42 following botulinum toxin type B injection (lower scores indicate more severe xerostomia). Conclusions: This study of patients with cervical dystonia suggests that

  17. PAAR-Rhs proteins harbor various C-terminal toxins to diversify the antibacterial pathways of type VI secretion systems.

    PubMed

    Ma, Jiale; Sun, Min; Dong, Wenyang; Pan, Zihao; Lu, Chengping; Yao, Huochun

    2017-01-01

    The type VI secretion system (T6SS) of bacteria plays a key role in competing for specific niches by the contact-dependent killing of competitors. Recently, Rhs proteins with polymorphic C-terminal toxin-domains that inhibit or kill neighboring cells were identified. In this report, we identified a novel Rhs with an MPTase4 (Metallopeptidase-4) domain (designated as Rhs-CT1) that showed an antibacterial effect via T6SS in Escherichia coli. We managed to develop a specific strategy by matching the diagnostic domain-architecture of Rhs-CT1 (Rhs with an N-terminal PAAR-motif and a C-terminal toxin domain) for effector retrieval and discovered a series of Rhs-CTs in E. coli. Indeed, the screened Rhs-CT3 with a REase-3 (Restriction endonuclease-3) domain also mediated interbacterial antagonism. Further analysis revealed that vgrGO1 and eagR/DUF1795 (upstream of rhs-ct) were required for the delivery of Rhs-CTs, suggesting eagR as a potential T6SS chaperone. In addition to chaperoned Rhs-CTs, neighborless Rhs-CTs could be classified into a distinct family (Rhs-Nb) sharing close evolutionary relationship with T6SS2-Rhs (encoded in the T6SS2 cluster of E. coli). Notably, the Rhs-Nb-CT5 was confirmed bioinformatically and experimentally to mediate interbacterial antagonism via Hcp2B-VgrG2 module. In a further retrieval analysis, we discovered various toxin/immunity pairs in extensive bacterial species that could be systematically classified into eight referential clans, suggesting that Rhs-CTs greatly diversify the antibacterial pathways of T6SS.

  18. GaAs laser treatment of bilateral eyelid ptosis due to complication of botulinum toxin type A injection.

    PubMed

    Majlesi, Gholamreza

    2008-10-01

    The widespread use of botulinum toxin type A (BTX-A) for aesthetic procedures in recent years has brought about some unwanted side effects that, though they are self-limited, cause inconvenience for patients. Injection of this paralytic toxin inactivates target muscle(s) for many months and unwanted facial movements will thus be prevented. Spreading of the toxin beyond the target muscles sometimes involves muscles necessary for other facial movements, such as the levator palpebrae, inactivation of which causes upper eyelid ptosis. Mild cases resolve after 2-3 wk, but in severe cases the complication may last as long as the cosmetic results persist (3-4 mo), and until now there has been no medical intervention to accelerate healing. In an effort to achieve more rapid recovery from eyelid ptosis due to overdose of BTX-A in the glabella, laser therapy was used in a 46-year-old woman with bilateral eyelid ptosis (partial on the right side and complete on the left) 12 d after injection. A GaAs laser was used and the protocol consisted of irradiation of three points on the upper lid just above the levator, and one point on the corrugator muscle on each side in contact mode, with three sessions per week (wavelength 890 nm, peak power 94 W, output power 28 mW, pulse duration 200 ns, spot size 3 mm, pulse repetition rate 3000 Hz, duration of irradiation 40 sec per point, energy per point 1.1 J, total energy per session 8.8 J, dose 16 J/cm2). The result was complete recovery from ptosis after 10 sessions, but the cosmetic results persisted for several months. It appears that if this procedure has similar results in other case series, it will be an effective therapeutic option to treat this complication.

  19. Sialidases Affect the Host Cell Adherence and Epsilon Toxin-Induced Cytotoxicity of Clostridium perfringens Type D Strain CN3718

    PubMed Central

    Li, Jihong; Sayeed, Sameera; Robertson, Susan; Chen, Jianming; McClane, Bruce A.

    2011-01-01

    Clostridium perfringens type B or D isolates, which cause enterotoxemias or enteritis in livestock, produce epsilon toxin (ETX). ETX is exceptionally potent, earning it a listing as a CDC class B select toxin. Most C. perfringens strains also express up to three different sialidases, although the possible contributions of those enzymes to type B or D pathogenesis remain unclear. Type D isolate CN3718 was found to carry two genes (nanI and nanJ) encoding secreted sialidases and one gene (nanH) encoding a cytoplasmic sialidase. Construction in CN3718 of single nanI, nanJ and nanH null mutants, as well as a nanI/nanJ double null mutant and a triple sialidase null mutant, identified NanI as the major secreted sialidase of this strain. Pretreating MDCK cells with NanI sialidase, or with culture supernatants of BMC206 (an isogenic CN3718 etx null mutant that still produces sialidases) enhanced the subsequent binding and cytotoxic effects of purified ETX. Complementation of BMC207 (an etx/nanH/nanI/nanJ null mutant) showed this effect is mainly attributable to NanI production. Contact between BMC206 and certain mammalian cells (e.g., enterocyte-like Caco-2 cells) resulted in more rapid sialidase production and this effect involved increased transcription of BMC206 nanI gene. BMC206 was shown to adhere to some (e.g. Caco-2 cells), but not all mammalian cells, and this effect was dependent upon sialidase, particularly NanI, expression. Finally, the sialidase activity of NanI (but not NanJ or NanH) could be enhanced by trypsin. Collectively these in vitro findings suggest that, during type D disease originating in the intestines, trypsin may activate NanI, which (in turn) could contribute to intestinal colonization by C. perfringens type D isolates and also increase ETX action. PMID:22174687

  20. Sialidases affect the host cell adherence and epsilon toxin-induced cytotoxicity of Clostridium perfringens type D strain CN3718.

    PubMed

    Li, Jihong; Sayeed, Sameera; Robertson, Susan; Chen, Jianming; McClane, Bruce A

    2011-12-01

    Clostridium perfringens type B or D isolates, which cause enterotoxemias or enteritis in livestock, produce epsilon toxin (ETX). ETX is exceptionally potent, earning it a listing as a CDC class B select toxin. Most C. perfringens strains also express up to three different sialidases, although the possible contributions of those enzymes to type B or D pathogenesis remain unclear. Type D isolate CN3718 was found to carry two genes (nanI and nanJ) encoding secreted sialidases and one gene (nanH) encoding a cytoplasmic sialidase. Construction in CN3718 of single nanI, nanJ and nanH null mutants, as well as a nanI/nanJ double null mutant and a triple sialidase null mutant, identified NanI as the major secreted sialidase of this strain. Pretreating MDCK cells with NanI sialidase, or with culture supernatants of BMC206 (an isogenic CN3718 etx null mutant that still produces sialidases) enhanced the subsequent binding and cytotoxic effects of purified ETX. Complementation of BMC207 (an etx/nanH/nanI/nanJ null mutant) showed this effect is mainly attributable to NanI production. Contact between BMC206 and certain mammalian cells (e.g., enterocyte-like Caco-2 cells) resulted in more rapid sialidase production and this effect involved increased transcription of BMC206 nanI gene. BMC206 was shown to adhere to some (e.g. Caco-2 cells), but not all mammalian cells, and this effect was dependent upon sialidase, particularly NanI, expression. Finally, the sialidase activity of NanI (but not NanJ or NanH) could be enhanced by trypsin. Collectively these in vitro findings suggest that, during type D disease originating in the intestines, trypsin may activate NanI, which (in turn) could contribute to intestinal colonization by C. perfringens type D isolates and also increase ETX action.

  1. The Role of Peganum harmala Ethanolic Extract and Type II Toxin Antitoxin System in Biofilm Formation.

    PubMed

    Valizadeh, Nasrin; Valian, Firuzeh; Sadeghifard, Nourkhoda; Karami, Shahriar; Pakzad, Iraj; Kazemian, Hossein; Ghafourian, Sobhan

    2017-03-20

    Toxin antitoxin system is a regulatory system that antitoxin inhibits the toxin. We aimed to determine the role of TA loci in biofilm formation in K. pneumoniae clinical and environmental isolates; also inhibition of biofilm formation by Peganum harmala. So, 40 K. pneumoniae clinical and environmental isolates were subjected for PCR to determine the frequency of mazEF, relEB, and mqsRA TA loci. Biofilm formation assay subjected for all isolates. Then, P. harmala was tested against positive biofilm formation strains. Our results demonstrated that relBE TA loci were dominant TA loci; whereas mqsRA TA loci were negative in all isolates. The most environmental isolates showed weak and no biofilm formation while strong and moderate biofilm formation observed in clinical isolates. Biofilm formations by K. pneumoniae in 9 ug/ml concentration were inhibited by P. harmala. In vivo study suggested to be performed to introduce Peganum harmala as anti-biofilm formation in K. pneumoniae.

  2. Botulinum toxin type A with oral baclofen versus oral tizanidine: a nonrandomized pilot comparison in patients with cerebral palsy and spastic equinus foot deformity.

    PubMed

    Dai, Alper I; Wasay, Mohammad; Awan, Safia

    2008-12-01

    The objective of this study was to compare the effectiveness of baclofen versus tizanidine as adjuvant treatment of botulinum toxin type A botulinum toxin type A in the management of children with spasticity. Thirty children with gastrocnemius spasticity were retrospectively reviewed at Gaziantep University Hospital, Gaziantep, Turkey. All patients were treated with localized botulinum toxin injections and baclofen or tizanidine for spasticity and were followed at 2- to 4-week intervals and evaluated for a total of 12 weeks; 17 children (57%) received baclofen and 13 (43%) received tizanidine. The mean score of Gross Motor Functional Measurement (76.63 +/- 5.88 vs 68.17 +/- 1.99; P < .001) and caregiver questionnaire scores (70.23 +/- 4.76 vs 66.59 +/- 3.53; P = .03) for the tizanidine group were significantly higher as compared with the baclofen group. This study suggests that combination of botulinum toxin type A with oral tizanidine is more effective with fewer side effects than combination of botulinum toxin type A and oral baclofen for spastic cerebral palsy.

  3. Confocal microscopy and exfoliative cytology

    PubMed Central

    Reddy, Shyam Prasad; Ramani, Pratibha; Nainani, Purshotam

    2013-01-01

    Context: Early detection of potentially malignant lesions and invasive squamous-cell carcinoma in the oral cavity could be greatly improved through techniques that permit visualization of subtle cellular changes indicative of the neoplastic transformation process. One such technique is confocal microscopy. Combining rapidity with reliability, an innovative idea has been put forward using confocal microscope in exfoliative cytology. Aims: The main objective of this study was to assess confocal microscopy for cytological diagnosis and the results were compared with that of the standard PAP stain. Settings and Design: Confocal microscope, acridine orange (AO) stain, PAP (Papanicolaou) stain. The study was designed to assess confocal microscopy for cytological diagnosis. In the process, smears of patients with (clinically diagnosed and/or suspected) oral squamous cell carcinoma as well as those of controls (normal people) were stained with acridine orange and observed under confocal microscope. The results were compared with those of the standard PAP method. Materials and Methods: Samples of buccal mucosa smears from normal patients and squamous cell carcinoma patients were made, fixed in 100% alcohol, followed by AO staining. The corresponding set of smears was stained with PAP stain using rapid PAP stain kit. The results obtained were compared with those obtained with AO confocal microscopy. Results: The study had shown nuclear changes (malignant cells) in the smears of squamous cell carcinoma patients as increased intensity of fluorescence of the nucleus, when observed under confocal microscope. Acridine orange confocal microscopy showed good amount of sensitivity and specificity (93%) in identifying malignant cells in exfoliative cytological smears. Conclusion: Confocal microscopy was found to have good sensitivity in the identification of cancer (malignant) cells in exfoliative cytology, at par with the PAP method. The rapidity of processing and screening a

  4. Confocal microscopy and exfoliative cytology.

    PubMed

    Reddy, Shyam Prasad; Ramani, Pratibha; Nainani, Purshotam

    2013-05-01

    Early detection of potentially malignant lesions and invasive squamous-cell carcinoma in the oral cavity could be greatly improved through techniques that permit visualization of subtle cellular changes indicative of the neoplastic transformation process. One such technique is confocal microscopy. Combining rapidity with reliability, an innovative idea has been put forward using confocal microscope in exfoliative cytology. The main objective of this study was to assess confocal microscopy for cytological diagnosis and the results were compared with that of the standard PAP stain. Confocal microscope, acridine orange (AO) stain, PAP (Papanicolaou) stain. The study was designed to assess confocal microscopy for cytological diagnosis. In the process, smears of patients with (clinically diagnosed and/or suspected) oral squamous cell carcinoma as well as those of controls (normal people) were stained with acridine orange and observed under confocal microscope. The results were compared with those of the standard PAP method. Samples of buccal mucosa smears from normal patients and squamous cell carcinoma patients were made, fixed in 100% alcohol, followed by AO staining. The corresponding set of smears was stained with PAP stain using rapid PAP stain kit. The results obtained were compared with those obtained with AO confocal microscopy. The study had shown nuclear changes (malignant cells) in the smears of squamous cell carcinoma patients as increased intensity of fluorescence of the nucleus, when observed under confocal microscope. Acridine orange confocal microscopy showed good amount of sensitivity and specificity (93%) in identifying malignant cells in exfoliative cytological smears. Confocal microscopy was found to have good sensitivity in the identification of cancer (malignant) cells in exfoliative cytology, at par with the PAP method. The rapidity of processing and screening a specimen resulted in saving of time. It added a certain amount of objectivity to the

  5. Efavirenz-induced exfoliative dermatitis.

    PubMed

    Zhang, Jiu-Cong; Sun, Yong-Tao

    2013-01-01

    Individuals with a human immunodeficiency virus (HIV) infection are at higher risk of developing adverse drug reactions. Multiple drugs are usually prescribed to patients with HIV infection for preventing the replication of HIV and for the treatment of the associated opportunistic infections. We report here the first case of an HIV-1-infected patient who developed an exfoliative dermatitis induced by efavirenz, a non-nucleoside reverse transcriptase inhibitor. Physicians should be aware of the possible occurrence of efavirenz-induced skin eruptions from the start of antiviral treatment of HIV infection.

  6. [Exfoliative esophagitis while taking dabigatran].

    PubMed

    Scheppach, Wolfgang; Meesmann, Malte

    2015-04-01

    History | A 77-year-old woman was admitted with severe chest pain, heartburn, dysphagia and odynophagia. She had been on dabigatran for 13 months due to atrial fibrillation and arterial hypertension. Investigations and findings | Endoscopy of the esophagus revealed sloughing of mucosal casts, predominantly in the upper half of the organ. Treatment and course | The patient was placed on pantoprazol, local anaesthetic antacid and i. v. fluids. Dabigatran was discontinued. The symptoms disappeared within 3 days. Control endoscopy after 12 days showed complete healing of the esophageal mucosa. Conclusion | The intake of dabigatran was associated with exfoliative esophagitis, possibly due to caustic tissue damage by prolonged drug contact.

  7. [Advances in the research of mechanism in prevention and treatment of scar with botulinum toxin type A and its clinical application].

    PubMed

    Li, Y H; Liu, J Q; Xiao, D; Zhang, W; Hu, D H

    2017-04-20

    Scar is a common complication in wound healing process, and how to effectively prevent and treat it is a hot and difficult problem in burns and plastic surgery field. Botulinum toxin type A is a neurotoxin that has been widely and effectively used in the cosmetic surgery field such as anti-wrinkle and thin face. In recent years, botulinum toxin type A has been applied in prevention and treatment of scar, which causes a great concern. Nowadays, the relevant reports have gradually increased, and the mechanisms have been explored more deeply. This article aims to summarize the possible mechanisms and clinical reports on the prevention and treatment of scar by botulinum toxin type A to provide a new way for the prevention and treatment of scar after surgery.

  8. The mycobacterial PhoH2 proteins are type II toxin antitoxins coupled to RNA helicase domains.

    PubMed

    Andrews, Emma S V; Arcus, Vickery L

    2015-07-01

    PhoH2 proteins are found in a diverse range of organisms that span the bacterial tree and little is known about this large protein family. PhoH2 proteins have two domains: An N-terminal PIN domain fused to a C-terminal PhoH domain. The genome of Mycobacterium tuberculosis encodes 48 PIN domains and 47 of these constitute the VapC components of the 47 VapBC toxin-antitoxins. The 48th member of the M. tuberculosis PIN domain array is found in the single PhoH2 protein encoded in the genome. All characterized PIN domain proteins are RNases and the PhoH domains are predicted ATPases. This fusion of a PIN domain with an ATPase reflects a much wider association between PIN domains and PhoH domains across many prokaryote genomes. Here, we examine PhoH2 proteins from M. tuberculosis, Mycobacterium smegmatis and a thermophilic homologue from Thermobispora bispora and we show that PhoH2 is a sequence-specific RNA helicase and RNAse. In addition, phoH2 from M. tuberculosis and M. smegmatis is part of a longer mRNA transcript which includes a small, unannotated open reading frame (ORF) upstream of the phoH2 gene. This small gene overlaps with the beginning of the phoH2 gene in a manner similar to the PIN domain toxin-antitoxin operons. We have annotated the upstream gene as phoAT and its putative promoter elements satisfy previously characterized consensus sequences at the -10 site. Conditional growth experiments carried out in M. smegmatis revealed a negative effect on growth by the expression of M. tuberculosis PhoH2 that was alleviated by co-expression of the PhoAT peptide. Thus in M. tuberculosis, PhoH2 represents a new variation on a type II PIN domain toxin-antitoxin systems such that the toxin-antitoxin is now coupled to an RNA helicase whose predicted biological function is to unwind and cleave RNA in a sequence specific manner.

  9. Interaction of three-finger toxins with phospholipid membranes: comparison of S- and P-type cytotoxins

    PubMed Central

    2004-01-01

    The CTs (cytotoxins) I and II are positively charged three-finger folded proteins from venom of Naja oxiana (the Central Asian cobra). They belong to S- and P-type respectively based on Ser-28 and Pro-30 residues within a putative phospholipid bilayer binding site. Previously, we investigated the interaction of CTII with multilamellar liposomes of dipalmitoylphosphatidylglycerol by wide-line 31P-NMR spectroscopy. To compare interactions of these proteins with phospholipids, we investigated the interaction of CTI with the multilamellar liposomes of dipalmitoylphosphatidylglycerol analogously. The effect of CTI on the chemical shielding anisotropy and deformation of the liposomes in the magnetic field was determined at different temperatures and lipid/protein ratios. It was found that both the proteins do not affect lipid organization in the gel state. In the liquid crystalline state of the bilayer they disturb lipid packing. To get insight into the interactions of the toxins with membranes, Monte Carlo simulations of CTI and CTII in the presence of the bilayer membrane were performed. It was found that both the toxins penetrate into the bilayer with the tips of all the three loops. However, the free-energy gain on membrane insertion of CTI is smaller (by ≈7 kcal/mol; 1 kcal≡4.184 kJ) when compared with CTII, because of the lower hydrophobicity of the membrane-binding site of CTI. These results clearly demonstrate that the P-type cytotoxins interact with membranes stronger than those of the S-type, although the mode of the membrane insertion is similar for both the types. PMID:15584897

  10. Separation medium containing thermally exfoliated graphite oxide

    NASA Technical Reports Server (NTRS)

    Prud'homme, Robert K. (Inventor); Aksay, Ilhan A. (Inventor); Herrera-Alonso, Margarita (Inventor)

    2012-01-01

    A separation medium, such as a chromatography filling or packing, containing a modified graphite oxide material, which is a thermally exfoliated graphite oxide with a surface area of from about 300 m.sup.2/g to 2600 m.sup.2/g, wherein the thermally exfoliated graphite oxide has a surface that has been at least partially functionalized.

  11. Influence of pH and temperature on the growth of and toxin production by neurotoxigenic strains of Clostridium butyricum type E.

    PubMed

    Anniballi, Fabrizio; Fenicia, Lucia; Franciosa, Giovanna; Aureli, Paolo

    2002-08-01

    Strains of Clostridium butyricum that produce botulinal toxin type E have been implicated in outbreaks of foodborne botulism in China, India, and Italy, yet the conditions that are favorable for the growth and toxinogenesis of these strains remain to be established. We attempted to determine the temperatures and pH levels that are most conducive to the growth of and toxin production by the six strains of neurotoxigenic C. butyricum that have been implicated in outbreaks of infective and foodborne botulism in Italy. The strains were cultured for 180 days on Trypticase-peptone-glucose-yeast extract broth at various pHs (4.6, 4.8, 5.0, 5.2, 5.4, 5.6, and 5.8) at 30 degrees C and at various temperatures (10, 12, and 15 degrees C) at pH 7.0. Growth was determined by checking for turbidity; toxin production was determined by the mouse bioassay. We also inoculated two foods: mascarpone cheese incubated at 25 and 15 degrees C and pesto sauce incubated at 25 degrees C. The lowest pH at which growth and toxin production occurred was 4.8 at 43 and 44 days of incubation, respectively. The lowest temperature at which growth and toxin production occurred was 12 degrees C, with growth and toxin production first being observed after 15 days. For both foods, toxin production was observed after 5 days at 25 degrees C. Since the strains did not show particularly psychrotrophic behavior, 4 degrees C can be considered a sufficiently low temperature for the inhibition of growth. However, the observation of toxin production in foods at room temperature and at abused refrigeration temperatures demands that these strains be considered a new risk for the food industry.

  12. Ouabain Protects Human Renal Cells against the Cytotoxic Effects of Shiga Toxin Type 2 and Subtilase Cytotoxin

    PubMed Central

    Amaral, María M.; Girard, Magalí C.; Álvarez, Romina S.; Paton, Adrienne W.; Paton, James C.; Repetto, Horacio A.; Sacerdoti, Flavia; Ibarra, Cristina A.

    2017-01-01

    Hemolytic uremic syndrome (HUS) is one of the most common causes of acute renal failure in children. The majority of cases are associated with Shiga toxin (Stx)-producing Escherichia coli (STEC). In Argentina, HUS is endemic and presents the highest incidence rate in the world. STEC strains expressing Stx type 2 (Stx2) are responsible for the most severe cases of this pathology. Subtilase cytotoxin (SubAB) is another STEC virulence factor that may contribute to HUS pathogenesis. To date, neither a licensed vaccine nor effective therapy for HUS is available for humans. Considering that Ouabain (OUA) may prevent the apoptosis process, in this study we evaluated if OUA is able to avoid the damage caused by Stx2 and SubAB on human glomerular endothelial cells (HGEC) and the human proximal tubule epithelial cell (HK-2) line. HGEC and HK-2 were pretreated with OUA and then incubated with the toxins. OUA protected the HGEC viability from Stx2 and SubAB cytotoxic effects, and also prevented the HK-2 viability from Stx2 effects. The protective action of OUA on HGEC and HK-2 was associated with a decrease in apoptosis and an increase in cell proliferation. Our data provide evidence that OUA could be considered as a therapeutic strategy to avoid the renal damage that precedes HUS. PMID:28718802

  13. Nanopore sensing of botulinum toxin type B by discriminating an enzymatically cleaved Peptide from a synaptic protein synaptobrevin 2 derivative.

    PubMed

    Wang, Yong; Montana, Vedrana; Grubišić, Vladimir; Stout, Randy F; Parpura, Vladimir; Gu, Li-Qun

    2015-01-14

    Botulinum neurotoxins (BoNTs) are the most lethal toxin known to human. Biodefense requires early and rapid detection of BoNTs. Traditionally, BoNTs can be detected by looking for signs of botulism in mice that receive an injection of human material, serum or stool. While the living animal assay remains the most sensitive approach, it is costly, slow and associated with legal and ethical constrains. Various biochemical, optical and mechanical methods have been developed for BoNTs detection with improved speed, but with lesser sensitivity. Here, we report a novel nanopore-based BoNT type B (BoNT-B) sensor that monitors the toxin's enzymatic activity on its substrate, a recombinant synaptic protein synaptobrevin 2 derivative. By analyzing the modulation of the pore current caused by the specific BoNT-B-digested peptide as a marker, the presence of BoNT-B at a subnanomolar concentration was identified within minutes. The nanopore detector would fill the niche for a much needed rapid and highly sensitive detection of neurotoxins, and provide an excellent system to explore biophysical mechanisms for biopolymer transportation.

  14. [Cloning and expression of ScFv gene against alpha-toxin of Clostridium perfringens type A].

    PubMed

    Zhao, B H; Xu, C B

    2001-09-01

    The VH and VL genes from a hybridoma cell line producing mouse McAb against alpha-toxin of Clostridium perfringens type A were amplified by RT-PCR. The VH and VL genes were connected thought a flexible linker (Gly4Ser)3 and the VH-linker-VL (ScFv) gene was cloned into a vector pGEM-T. The ScFv gene consists of 726 bp encoding 242 amino acid residues. Both VH and VL genes were confirmed as functionally rearranged mouse immunoglobulin variable region. According to kabat classed method, the VH and VL gene segments belong to mouse Ig heavy chain subgroup II (B) and kappa light chain subgroup III respectively. The ScFv gene was amplified inserted the expression vector pHOG21 and transformed into E coli XL1-BLUE. The ScFv protein was highly expressed in recombinant strain XL1-BLUE (pHOG-2E3) and the expression level of the ScFv was about 25% of total bacteria protein by SDS-PAGE. The neutralization assay showed that the expressed ScFv protein could neutralize the phospholipase C activities of alpha-toxin.

  15. Hydrogen Selective Exfoliated Zeolite Membranes

    SciTech Connect

    Tsapatsis, Michael; Daoutidis, Prodromos; Elyassi, Bahman; Lima, Fernando; Iyer, Aparna; Agrawal, Kumar; Sabnis, Sanket

    2015-04-06

    The objective of this project was to develop and evaluate an innovative membrane technology at process conditions that would be representative of Integrated Gasification Combined Cycle (IGCC) advanced power generation with pre-combustion capture of carbon dioxide (CO2). This research focused on hydrogen (H2)-selective zeolite membranes that could be utilized to separate conditioned syngas into H2-rich and CO2-rich components. Both experiments and process design and optimization calculations were performed to evaluate the concept of ultra-thin membranes made from zeolites nanosheets. In this work, efforts in the laboratory were made to tackle two fundamental challenges in application of zeolite membranes in harsh industrial environments, namely, membrane thickness and membrane stability. Conventional zeolite membranes have thicknesses in the micron range, limiting their performance. In this research, we developed a method for fabrication of ultimately thin zeolite membranes based on zeolite nanosheets. A range of layered zeolites (MWW, RWR, NSI structure types) suitable for hydrogen separation was successfully exfoliated to their constituent nanosheets. Further, membranes were made from one of these zeolites, MWW, to demonstrate the potential of this group of materials. Moreover, long-term steam stability of these zeolites (up to 6 months) was investigated in high concentrations of steam (35 mol% and 95 mole%), high pressure (10 barg), and high temperatures (350 °C and 600 °C) relevant to conditions of water-gas-shift and steam methane reforming reactions. It was found that certain nanosheets are stable, and that stability depends on the concentration of structural defects. Additionally, models that represent a water-gas-shift (WGS) membrane reactor equipped with the zeolite membrane were developed for systems studies. These studies had the aim of analyzing the effect of the membrane reactor integration into IGCC plants

  16. [Treatment of spasticity in nursing homes: botulinum toxin type A as part of therapy].

    PubMed

    Wolswijk, Adrie H M; Dirkx, Anita E M

    2015-01-01

    Complications of spasticity can severely limit daily activities and care-giving. For those who treat or provide care to patients with spasticity in nursing homes, it is important to recognise complaints in order to prevent serious complications such as care-related pain, contractures and pressure sores. The involvement of a rehabilitation physician is essential to provide a high standard of care. We present two nursing home patients, a 95-year-old woman and a 63-year-old man, with severe upper limb complications following spasticity. Both patients received botulinum toxin injections in the affected muscles, combined with an appropriate splint. A treatment team consisting of a specialist in geriatric medicine, a rehabilitation physician, a physical and an occupational therapist provided consistent daily care in the institution. These efforts substantially reduced care-related pain and improved social behaviour and care options. If spasticity prohibits treatment or care, consultation of a rehabilitation physician at an early stage is indicated.

  17. Botulinum toxin type-A in the management of spastic equinovarus deformity after stroke

    PubMed Central

    Otom, Ali H.; Al-Khawaja, Imad M.; Al-Quliti, Khalid W.

    2014-01-01

    Objective: To retrospectively compare 2 injection techniques in the management of spastic equinovarus deformity after stroke. Methods: Patients with stroke were seen at King Hussein Medical Center, Amman, Jordan between January and December 2009. The study design involved an open label retrospective analysis of medical records of 2 groups of comparable age and onset of first stroke. Botulinum toxin was injected into the calf muscles at 2 sites in group I (12 patients) and 4 sites in group II (14 patients). Functional gain was evaluated by the time to walk 10 meters at month one, 3, and 6 compared with baseline. Results: There was significant improvement in walking time in each study group. However, there was no significant difference between the 2 groups as measured by the 10-meter walking time. Conclusion: Fewer injection sites would minimize patient discomfort and possibly the production of antibodies, yielding similar therapeutic effects. PMID:24983281

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

    PubMed Central

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

    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 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

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

    PubMed

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

    2015-09-23

    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.

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

    PubMed

    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.

  1. Hypothesis: type I toxin-antitoxin genes enter the persistence field-a feedback mechanism explaining membrane homoeostasis.

    PubMed

    Gerdes, Kenn

    2016-11-05

    Bacteria form persisters, cells that are tolerant to multiple antibiotics and other types of environmental stress. Persister formation can be induced either stochastically in single cells of a growing bacterial ensemble, or by environmental stresses, such as nutrient starvation, in a subpopulation of cells. In many cases, the molecular mechanisms underlying persistence are still unknown. However, there is growing evidence that, in enterobacteria, both stochastically and environmentally induced persistence are controlled by the second messenger (p)ppGpp. For example, the 'alarmone' (p)ppGpp activates Lon, which, in turn, activates type II toxin-antitoxin (TA) modules to thereby induce persistence. Recently, it has been shown that a type I TA module, hokB/sokB, also can induce persistence. In this case, the underlying mechanism depends on the universally conserved GTPase Obg and, surprisingly, also (p)ppGpp. In the presence of (p)ppGpp, Obg stimulates hokB transcription and induces persistence. HokB toxin expression is under both negative and positive control: SokB antisense RNA inhibits hokB mRNA translation, while (p)ppGpp and Obg together stimulate hokB transcription. HokB is a small toxic membrane protein that, when produced in modest amounts, leads to membrane depolarization, cell stasis and persistence. By contrast, overexpression of HokB disrupts the membrane potential and kills the cell. These observations raise the question of how expression of HokB is regulated. Here, I propose a homoeostatic control mechanism that couples HokB expression to the membrane-bound RNase E that degrades and inactivates SokB antisense RNA.This article is part of the themed issue 'The new bacteriology'. © 2016 The Author(s).

  2. [Characterization of a Cl. Perfringens type D strain, isolated in the field and optimization of epsilon toxin biosynthesis in a cell culture].

    PubMed

    Maaroufi, A; Metoui, W; Rahmouni, S; Ghram, A

    2000-01-01

    A field strain of cl. perfringens, named Dt001, was isolated from kidney of ovine enterotoemia case. The isolate characterized as Cl. perfringens, type D was based on its cultural and biochemical characters and its factors of virulence. The strain was very toxinogenic and well adapted to culture conditions of biofermentation when the parameters related to ptt, incubation time, substrat ... were optimized. Thus, the use of carbon source as polymer (destrine), the continuous control of pH allowed improvement of the rate of biosynthesis of Epsilon toxine by 10 times. The study of the immunogenicity of the isolate showed that preparations of anacultures were more immunogenic then those of anatoxine type. The fact that the two forms of epsilon antigens (protoxin and active toxin) show similar immune response in rabbits, indicates that the proteolytic action of trypsin is limited only to the toxic sites and does not affect the immunogenic epsitopes of the toxin. It also suggests a molecular organization of epsilon toxin in which the immunogenic epsitopes and the toxin sites are apart. The biotechnological performances and the immunogenicity and toxinogenical of the Dt001 isolate are in favor of its possible use as a component of an inactivated vaccine against enterotoxenia.

  3. Cloning and expression of Clostridium perfringens type D vaccine strain epsilon toxin gene in E. coli as a recombinant vaccine candidate

    PubMed Central

    Aziminia, Parastoo; Pilehchian-Langroudi, Reza; Esmaeilnia, Kasra

    2016-01-01

    Background and Objectives: Clostridium perfringens, a Gram-positive obligate anaerobic bacterium, is able to form resistant spores which are widely distributed in the environment. C. perfringens is subdivided into five types A to E based on its four major alpha, beta, epsilon and iota toxins. The aim of the present study was cloning and expression of C. perfringens type D vaccine strain epsilon toxin gene. Materials and Methods: Genomic DNA was extracted and the epsilon toxin gene was amplified using Pfu DNA polymerase. The PCR product was cloned into pJET1.2/blunt cloning vector. The recombinant vector (pJETε) was sequenced using universal primers. At the next step epsilon toxin gene was subcloned into pET22b(+) expression vector and transformed into E. coli Rosetta (DE3) host strain. Results: The recombinant protein has been expressed in E. coli Rosetta (DE3) cells after subcloning of C. perfringens etx gene (1008 bp) into the expression vector. Conclusion: We concluded that E. coli Rosetta strain was suitable for the expression of recombinant C. perfringens epsilon toxin protein from pET22ε expression vector. This recombinant cell can be used for further research on recombinant vaccine development. PMID:28210460

  4. Paroxysmal Autonomic Instability With Dystonia Managed Using Chemodenervation Including Alcohol Neurolysis and Botulinum Toxin Type A Injection: A Case Report

    PubMed Central

    Lee, Hye-Sun; Oh, Hyun-Seung

    2015-01-01

    Paroxysmal autonomic instability with dystonia (PAID) is a rare complication of brain injury. Symptoms of PAID include diaphoresis, hyperthermia, hypertension, tachycardia, and tachypnea accompanied by hypertonic movement. Herein, we present the case of a 44-year-old female patient, who was diagnosed with paraneoplastic limbic encephalopathy caused by thyroid papillary cancer. The patient exhibited all the symptoms of PAID. On the basis that the symptoms were unresponsive to antispastic medication and her liver function test was elevated, we performed alcohol neurolysis of the musculocutaneous nerve followed by botulinum toxin type A (BNT-A) injection into the biceps brachii and brachialis. Unstable vital signs and hypertonia were relieved after chemodenervation. Accordingly, alcohol neurolysis and BNT-A injection are proposed as a treatment option for intractable PAID. PMID:25932429

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

    NASA Astrophysics Data System (ADS)

    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.

  6. Botulinum Toxin Type-A (BoNT-A) Injections of the Corrugator Muscles for Aesthetics and Depression?

    PubMed

    Brennan, Connie

    The treatment of glabellar lines with botulinum toxin type-A (BoNT-A) is a staple for aesthetic providers who specialize in facial rejuvenation. Clinical efforts are currently underway to substantiate upper facial injections (the corrugator muscles are the target muscles) of BoNT-A as an antidepression therapy. This article describes the origin of "facial feedback" by Charles Darwin nearly 150 years ago, as well as "emotional proprioception"-2 neuroanatomical concepts that help provide the scientific rationale behind the general influence facial muscles have on the emotional centers of the brain, and, specifically, how the corrugator muscles-involved with frowning-promote a gloomy mood. The journey researchers have taken to clinically qualify BoNT-A injections of the corrugator muscles-to inhibit frowning, and thereby mitigate depression-will also be highlighted so that aesthetic providers are up to date on this emerging benefit of BoNT-A.

  7. Graphene reflux: improving the yield of liquid-exfoliated nanosheets through repeated separation techniques

    NASA Astrophysics Data System (ADS)

    Rountree, Kyler S.; Shah, Smit A.; Sweeney, Charles B.; Irin, Fahmida; Green, Micah J.

    2016-12-01

    Scalable production of graphene through liquid-phase exfoliation has been plagued by low yields. Although several recent studies have attempted to improve graphene exfoliation technology, the problem of separating colloidal nanosheets from unexfoliated parent material has received far less attention. Here we demonstrate a scalable method for improving nanosheet yield through a facile washing process. By probing the sedimentation of liquid-phase exfoliated slurries of graphene nanosheets and parent material, we found that a portion of exfoliated graphene is entrapped in the sediment, but can be recovered by repeatedly washing the slurry of nanosheet and parent material with additional solvent. We found this process to significantly increase the overall yield of graphene (graphene/parent material) and recover a roughly constant proportion of graphene with each wash. The cumulative amount of graphene recovered is only a function of total solvent volume. Moreover, we found this technique to be applicable to other types of nanosheets such as boron nitride nanosheets.

  8. A study of synchronization of quantal transmitter release from mammalian motor endings by the use of botulinal toxins type A and D.

    PubMed Central

    Molgó, J; Siegel, L S; Tabti, N; Thesleff, S

    1989-01-01

    1. The effects of botulinum toxin (BoTx) types A and D on spontaneous and evoked phasic transmitter release were studied in the isolated extensor digitorum longus muscle of the rat or the levator auris longus muscle of mice. 2. The toxins were injected subcutaneously into the hindleg of adult rats or the dorsal aspect of the neck of mice. At various times after the injection the muscles were removed from the anaesthetized animal and neuromuscular transmission examined in vitro by conventional intracellular techniques. 3. Both toxins reduced spontaneous transmitter release recorded as the frequency of miniature end-plate potentials but BoTx type D was less effective in that respect than the type A toxin. 4. With both toxins the block of evoked phasic transmitter release, recorded as end-plate potentials, was almost complete. As previously reviewed by Simpson (1986) the block produced by BoTx type A was partially reversed by procedures which elevate the intraterminal level of calcium ions. However, in BoTx type D-paralysed muscles such procedures failed to restore phasic transmitter release but caused a period of high-frequency asynchronous transmitter release following each nerve impulse. 5. To investigate if the lack of synchronization of evoked transmitter release observed in BoTx type D-paralysed muscles was due to alterations in presynaptic currents we examined, by perineural recordings, the Na+, fast K+, slow K+, K+-Ca2+-dependent and the Ca2+ currents in BoTx type D-paralysed muscles. These presynaptic currents were not altered as compared to unpoisoned controls. 6. We suggest that there exists a presynaptic process, which in addition to Ca2+ influx participates in transmitter synchronization and which is a main target for BoTx type D action. PMID:2575665

  9. Botulinum Toxin Type A Injections for Cervical and Shoulder Girdle Myofascial Pain Using an Enriched Protocol Design

    PubMed Central

    Nicol, Andrea L.; Wu, Irene I.; Ferrante, F. Michael

    2014-01-01

    Background Myofascial pain syndrome is a regional condition of muscle pain and stiffness and is classically characterized by the presence of trigger points in affected musculature. Botulinum toxin type A (BoNT-A) has been shown to have antinociceptive properties and elicit sustained muscle relaxation, thereby possibly affording even greater relief than traditional strategies. Our goal in this study was to determine whether direct injection of BoNT-A into painful muscle groups is effective for cervical and shoulder girdle myofascial pain. Methods An enriched protocol design was used wherein 114 patients with cervical and shoulder girdle myofascial pain underwent injection of BoNT-A to determine their response to the drug. Fifty-four responders were then enrolled in a twelve-week, randomized, double-blind, placebo-controlled trial. Pain scales and quality of life measures were assessed at baseline and at routine follow-up visits until completion of the study after 26 weeks. Results Injection of BoNT-A into painful muscle groups improved average visual numerical pain scores in subjects who received a second dose of BoNT-A compared to placebo (p = 0.019 (0.26, 2.78)). Subjects who received a second dose of BoNT-A had a reduced number of headaches per week (p = 0.04 (0.07, 4.55)). Brief Pain Inventory interference scores for general activity and sleep were improved (p = 0.046 (0.038, 3.7) and 0.02 (0.37, 4.33), respectively) in those who received a second dose of BoNT-A. Conclusion Botulinum toxin type A injected directly into painful muscle groups improves average pain scores and certain aspects of quality of life in patients suffering from severe cervical and shoulder girdle myofascial pain. PMID:24842179

  10. Distinct Physiologic and Inflammatory Responses Elicited in Baboons after Challenge with Shiga Toxin Type 1 or 2 from Enterohemorrhagic Escherichia coli▿

    PubMed Central

    Stearns-Kurosawa, D. J.; Collins, Valta; Freeman, Scott; Tesh, Vernon L.; Kurosawa, Shinichiro

    2010-01-01

    Shiga toxin-producing Escherichia coli is a principal source of regional outbreaks of bloody diarrhea and hemolytic-uremic syndrome in the United States and worldwide. Primary bacterial virulence factors are Shiga toxin types 1 and 2 (Stx1 and Stx2), and we performed parallel analyses of the pathophysiologies elicited by the toxins in nonhuman primate models to identify shared and unique consequences of the toxemias. After a single intravenous challenge with purified Stx1 or Stx2, baboons (Papio) developed thrombocytopenia, anemia, and acute renal failure with loss of glomerular function, in a dose-dependent manner. Differences in the timing and magnitude of physiologic responses were observed between the toxins. The animals were more sensitive to Stx2, with mortality at lower doses, but Stx2-induced renal injury and mortality were delayed 2 to 3 days compared to those after Stx1 challenge. Multiplex analyses of plasma inflammatory cytokines revealed similarities (macrophage chemoattractant protein 1 [MCP-1] and tumor necrosis factor alpha [TNF-α]) and differences (interleukin-6 [IL-6] and granulocyte colony-stimulating factor [G-CSF]) elicited by the toxins with respect to the mediator induced and timing of the responses. Neither toxin induced detectable levels of plasma TNF-α. To our knowledge, this is the first time that the in vivo consequences of the toxins have been compared in a parallel and reproducible manner in nonhuman primates, and the data show similarities to patient observations. The availability of experimental nonhuman primate models for Stx toxemias provides a reproducible platform for testing antitoxin compounds and immunotherapeutics with outcome criteria that have clinical meaning. PMID:20308301

  11. Indicators: Algal Toxins (microcystin)

    EPA Pesticide Factsheets

    Algal toxins are toxic substances released by some types of algae (phytoplankton) when they are present in large quantities (blooms) and decay or degrade. High nutrient levels and warm temperatures often result in favorable conditions for algae blooms.

  12. The effect of spasticity on cortical somatosensory-evoked potentials: changes of cortical somatosensory-evoked potentials after botulinum toxin type A injection.

    PubMed

    Park, Eun Sook; Park, Chang Il; Kim, Deog Young; Kim, Yong Rae

    2002-11-01

    To evaluate the changes in cortical somatosensory-evoked potentials (SEPs) after botulinum toxin type A injection to determine what effect spasticity has on cortical SEPs. Intervention study and before-after trial. University-affiliated hospital in Korea. Twelve children with spastic hemiplegic cerebral palsy (CP), 7 children with spastic diplegic CP, and 8 patients with traumatic brain injury. All participants had botulinum toxin type A injected into the muscles of the spastic limb. SEPs were recorded before and 7 days after the botulinum toxin type A injection. Spasticity of the affected spastic limb was also measured. The short latency and amplitude of waves in SEPs were measured. The SEP results were divided into 3 groups: flat (no evoked potential), abnormal (evoked but delayed in latency), and normal (clear waveform with normal latency). The normal response of cortical SEP increased after injection. The SEPs exhibited more frequent improvement in the limbs, with greater improvement of spasticity in grade (>1.0 grade) and in patients of younger age (<3y) after injection (P<.05). The observed improvement of cortical SEPs with associated reduction of spasticity that occurred after the botulinum toxin type A injection indicates that spasticity itself can be considered a factor affecting cortical SEPs. Copyright 2002 by the American Congress of Rehabilitation Medicine and the American Academy of Physical Medicine and Rehabilitation

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

    ERIC Educational Resources Information Center

    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…

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

    ERIC Educational Resources Information Center

    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…

  15. Genome sequencing and comparative genomics provides insights on the evolutionary dynamics and pathogenic potential of different H-Types of Shiga toxin-producing Escherichia coli O104

    USDA-ARS?s Scientific Manuscript database

    Various Shiga toxin-producing Escherichia coli (STEC) O104 H-types including H4, H7, H21, and H¯ have been associated with sporadic cases of illness and have caused outbreaks globally. In the U.S., STEC O104:H21 caused an outbreak associated with milk in 1994. The aim of this work was to conduct a...

  16. Protection against Shiga Toxins

    PubMed Central

    Kavaliauskiene, Simona; Dyve Lingelem, Anne Berit; Skotland, Tore; Sandvig, Kirsten

    2017-01-01

    Shiga toxins consist of an A-moiety and five B-moieties able to bind the neutral glycosphingolipid globotriaosylceramide (Gb3) on the cell surface. To intoxicate cells efficiently, the toxin A-moiety has to be cleaved by furin and transported retrogradely to the Golgi apparatus and to the endoplasmic reticulum. The enzymatically active part of the A-moiety is then translocated to the cytosol, where it inhibits protein synthesis and in some cell types induces apoptosis. Protection of cells can be provided either by inhibiting binding of the toxin to cells or by interfering with any of the subsequent steps required for its toxic effect. In this article we provide a brief overview of the interaction of Shiga toxins with cells, describe some compounds and conditions found to protect cells against Shiga toxins, and discuss whether they might also provide protection in animals and humans. PMID:28165371

  17. Comparison of Efficacy and Side Effects of Oral Baclofen Versus Tizanidine Therapy with Adjuvant Botulinum Toxin Type A in Children With Cerebral Palsy and Spastic Equinus Foot Deformity.

    PubMed

    Dai, Alper I; Aksoy, Sefika N; Demiryürek, Abdullah T

    2016-02-01

    This retrospective study aimed to compare the therapeutic response, including side effects, for oral baclofen versus oral tizanidine therapy with adjuvant botulinum toxin type A in a group of 64 pediatric patients diagnosed with static encephalopathy and spastic equinus foot deformity. Following botulinum toxin A treatment, clinical improvement led to the gradual reduction of baclofen or tizanidine dosing to one-third of the former dose. Gross Motor Functional Measure and Caregiver Health Questionnaire scores were markedly elevated post-botulinum toxin A treatment, with scores for the tizanidine (Gross Motor Functional Measure: 74.45 ± 3.72; Caregiver Health Questionnaire: 72.43 ± 4.29) group significantly higher than for the baclofen group (Gross Motor Functional Measure: 68.23 ± 2.66; Caregiver Health Questionnaire: 67.53 ± 2.67, P < .001). These findings suggest that the combined use of botulinum toxin A and a low dose of tizanidine in treating children with cerebral palsy appears to be more effective and has fewer side effects versus baclofen with adjuvant botulinum toxin A.

  18. Phospholipase C produced by Clostridium botulinum types C and D: comparison of gene, enzymatic, and biological activities with those of Clostridium perfringens alpha-toxin.

    PubMed

    Fatmawati, Ni Nengah Dwi; Sakaguchi, Yoshihiko; Suzuki, Tomonori; Oda, Masataka; Shimizu, Kenta; Yamamoto, Yumiko; Sakurai, Jun; Matsushita, Osamu; Oguma, Keiji

    2013-01-01

    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.

  19. Biological activity of two botulinum toxin type A complexes (Dysport and Botox) in volunteers: a double-blind, randomized, dose-ranging study.

    PubMed

    Wohlfarth, K; Schwandt, I; Wegner, F; Jürgens, T; Gelbrich, G; Wagner, A; Bogdahn, U; Schulte-Mattler, W

    2008-12-01

    Despite extensive clinical experience and published data regarding botulinum toxin, questions remain about the clinical substitution of one botulinum toxin formulation for another. In the case of Dysport and Botox, dose-equivalence ratios ranging from 1:1 to 6:1 (Dysport:Botox) have been advocated. This dose-ranging, electroneurographic study investigated the dose equivalence, diffusion characteristics (spread) and safety of these two type-A toxins in 79 volunteers. Dysport and Botox caused significant and similar reductions in compound muscle action potential (CMAP) amplitude in the target muscle (extensor digitorum brevis, EDB) 2 weeks after injection, with effects persisting to the 12-week timepoint. For both products, the reduction in amplitude was increased with increasing doses and with increasing concentration. The effects of toxin on neighbouring muscles were much smaller and of a shorter duration than those on the target muscle, implying a modest spread of toxin. Unlike the target muscle, the effects were greater with the higher volume, suggesting this volume led to greater diffusion from the EDB. No adverse events were reported. Statistical modelling with CMAP amplitude data from the target muscle gave a bioequivalence of 1.57 units of Dysport:1 unit of Botox (95 % CI: 0.77-3.20 units). The data indicate that a dose-equivalence ratio of 3:1 was within the statistical error limits, but ratios over 3:1 are too high.

  20. New secreted toxins and immunity proteins encoded within the Type VI secretion system gene cluster of Serratia marcescens

    PubMed Central

    English, Grant; Trunk, Katharina; Rao, Vincenzo A; Srikannathasan, Velupillai; Hunter, William N; Coulthurst, Sarah J

    2012-01-01

    Protein secretion systems are critical to bacterial virulence and interactions with other organisms. The Type VI secretion system (T6SS) is found in many bacterial species and is used to target either eukaryotic cells or competitor bacteria. However, T6SS-secreted proteins have proven surprisingly elusive. Here, we identified two secreted substrates of the antibacterial T6SS from the opportunistic human pathogen, Serratia marcescens. Ssp1 and Ssp2, both encoded within the T6SS gene cluster, were confirmed as antibacterial toxins delivered by the T6SS. Four related proteins encoded around the Ssp proteins (‘Rap’ proteins) included two specifically conferring self-resistance (‘immunity’) against T6SS-dependent Ssp1 or Ssp2 toxicity. Biochemical characterization revealed specific, tight binding between cognate Ssp–Rap pairs, forming complexes of 2:2 stoichiometry. The atomic structures of two Rap proteins were solved, revealing a novel helical fold, dependent on a structural disulphide bond, a structural feature consistent with their functional localization. Homologues of the Serratia Ssp and Rap proteins are found encoded together within other T6SS gene clusters, thus they represent founder members of new families of T6SS-secreted and cognate immunity proteins. We suggest that Ssp proteins are the original substrates of the S. marcescens T6SS, before horizontal acquisition of other T6SS-secreted toxins. Molecular insight has been provided into how pathogens utilize antibacterial T6SSs to overcome competitors and succeed in polymicrobial niches. PMID:22957938

  1. Natural killer T (NKT) cells accelerate Shiga toxin type 2 (Stx2) pathology in mice

    PubMed Central

    Obata, Fumiko; Subrahmanyam, Priyanka B.; Vozenilek, Aimee E.; Hippler, Lauren M.; Jeffers, Tynae; Tongsuk, Methinee; Tiper, Irina; Saha, Progyaparamita; Jandhyala, Dakshina M.; Kolling, Glynis L.; Latinovic, Olga; Webb, Tonya J.

    2015-01-01

    Shiga toxin-producing Escherichia coli (STEC) is a leading cause of childhood renal disease Hemolytic Uremic Syndrome (HUS). The involvement of renal cytokines and chemokines is suspected to play a critical role in disease progression. In current article, we tested the hypothesis that NKT cells are involved in Stx2-induced pathology in vivo. To address this hypothesis we compared Stx2 toxicity in WT and CD1 knockout (KO) mice. In CD1KO mice, which lack natural killer T (NKT) cells, Stx2-induced pathologies such as weight loss, renal failure, and death were delayed. In WT mice, Stx2-specific selective increase in urinary albumin occurs in later time points, and this was also delayed in NKT cell deficient mice. NKT cell-associated cytokines such as IL-2, IL-4, IFN-γ, and IL-17 were detected in kidney lysates of Stx2-injected WT mice with the peak around 36 h after Stx2 injection. In CD1KO, there was a delay in the kinetics, and increases in these cytokines were observed 60 h post Stx2 injection. These data suggest that NKT cells accelerate Stx2-induced pathology in mouse kidneys. To determine the mechanism by which NKT cells promote Stx2-associated disease, in vitro studies were performed using murine renal cells. We found that murine glomerular endothelial cells and podocytes express functional CD1d molecules and can present exogenous antigen to NKT cells. Moreover, we observed the direct interaction between Stx2 and the receptor Gb3 on the surface of mouse renal cells by 3D STORM-TIRF which provides single molecule imaging. Collectively, these data suggest that Stx2 binds to Gb3 on renal cells and leads to aberrant CD1d-mediated NKT cell activation. Therefore, strategies targeting NKT cells could have a significant impact on Stx2-associated renal pathology in STEC disease. PMID:25904903

  2. The Accessory Genome of Shiga Toxin-Producing Escherichia coli Defines a Persistent Colonization Type in Cattle

    PubMed Central

    Barth, Stefanie A.; Menge, Christian; Eichhorn, Inga; Semmler, Torsten; Wieler, Lothar H.; Pickard, Derek; Belka, Ariane; Berens, Christian

    2016-01-01

    ABSTRACT Shiga toxin-producing Escherichia coli (STEC) strains can colonize cattle for several months and may, thus, serve as gene reservoirs for the genesis of highly virulent zoonotic enterohemorrhagic E. coli (EHEC). Attempts to reduce the human risk for acquiring EHEC infections should include strategies to control such STEC strains persisting in cattle. We therefore aimed to identify genetic patterns associated with the STEC colonization type in the bovine host. We included 88 persistent colonizing STEC (STECper) (shedding for ≥4 months) and 74 sporadically colonizing STEC (STECspo) (shedding for ≤2 months) isolates from cattle and 16 bovine STEC isolates with unknown colonization types. Genoserotypes and multilocus sequence types (MLSTs) were determined, and the isolates were probed with a DNA microarray for virulence-associated genes (VAGs). All STECper isolates belonged to only four genoserotypes (O26:H11, O156:H25, O165:H25, O182:H25), which formed three genetic clusters (ST21/396/1705, ST300/688, ST119). In contrast, STECspo isolates were scattered among 28 genoserotypes and 30 MLSTs, with O157:H7 (ST11) and O6:H49 (ST1079) being the most prevalent. The microarray analysis identified 139 unique gene patterns that clustered with the genoserotypes and MLSTs of the strains. While the STECper isolates possessed heterogeneous phylogenetic backgrounds, the accessory genome clustered these isolates together, separating them from the STECspo isolates. Given the vast genetic heterogeneity of bovine STEC strains, defining the genetic patterns distinguishing STECper from STECspo isolates will facilitate the targeted design of new intervention strategies to counteract these zoonotic pathogens at the farm level. IMPORTANCE Ruminants, especially cattle, are sources of food-borne infections by Shiga toxin-producing Escherichia coli (STEC) in humans. Some STEC strains persist in cattle for longer periods of time, while others are detected only sporadically

  3. Global Aesthetics Consensus: Botulinum Toxin Type A—Evidence-Based Review, Emerging Concepts, and Consensus Recommendations for Aesthetic Use, Including Updates on Complications

    PubMed Central

    Signorini, Massimo; Liew, Steven; Trindade de Almeida, Ada R.; Wu, Yan; Vieira Braz, André; Fagien, Steven; Goodman, Greg J.; Monheit, Gary; Raspaldo, Hervé

    2016-01-01

    Background: Botulinum toxin type A injection remains the leading nonsurgical cosmetic procedure worldwide, with a high rate of efficacy and patient satisfaction. Methods: A multinational, multidisciplinary group of plastic surgeons and dermatologists convened the Global Aesthetics Consensus Group to develop updated consensus recommendations with a worldwide perspective for botulinum toxin and hyaluronic acid fillers. This publication on botulinum toxin type A considers advances in facial analysis, injection techniques, and avoidance and management of complications. Results: Use of botulinum toxin has evolved from the upper face to also encompass the lower face, neck, and midface. The Global Aesthetics Consensus Group emphasizes an integrative, diagnostic approach. Injection dosage and placement are based on analysis of target muscles in the context of adjacent ones and associated soft and hard tissues. The indication for selection of botulinum toxin as a primary intervention is that excessive muscular contraction is the primary etiology of the facial disharmony to be addressed. Global Aesthetics Consensus Group recommendations demonstrate a paradigm shift toward neuromodulation rather than paralysis, including lower dosing of the upper face, more frequent combination treatment with hyaluronic acid fillers, and intracutaneous injection where indicated to limit depth and degree of action. Conclusions: The accumulation of clinical evidence and experience with botulinum toxin has led to refinements in treatment planning and implementation. The Global Aesthetics Consensus Group advocates an etiology-driven, patient-tailored approach, to enable achievement of optimal efficacy and safety in patient populations that are rapidly diversifying with respect to ethnicity, gender, and age. CLINCAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V. PMID:26910696

  4. BmTx3, a scorpion toxin with two putative functional faces separately active on A-type K+ and HERG currents.

    PubMed Central

    Huys, Isabelle; Xu, Chen-Qi; Wang, Cheng-Zhong; Vacher, Hélène; Martin-Eauclaire, Marie-France; Chi, Cheng-Wu; Tytgat, Jan

    2004-01-01

    A novel HERG channel blocker was isolated from the venom of the scorpion Buthus martensi Karsch, sequenced and characterized at the pharmacological level after chemical synthesis. According to the determined amino acid sequence, the cDNA and genomic genes were then cloned. The genomic gene consists of two exons interrupted by an intron of 65 bp at position -6 upstream from the mature toxin. The protein sequence of this toxin was completely identical with that of a known A-type K+ current blocker BmTx3, belonging to scorpion alpha-KTx subfamily 15. Thus BmTx3 is the first reported alpha-KTx peptide also showing HERG-blocking activity, like gamma-KTx peptides. Moreover, different from classical alpha-KTx peptides, such as charybdotoxin, BmTx3 cannot block Shaker -type K+ channels. Phylogenetic tree analysis reveals that this toxin takes an intermediate position between classical alpha-KTx and gamma-KTx toxins. From a structural point of view, we propose that two separate functional faces might exist on the BmTx3 molecule, responsible for the two different K+-current-blocking functions. Face A, composed of Arg18 and Lys19 in the alpha-helix side, might correspond to HERG blocking activity, whereas Face B, containing a putative functional dyad (Lys27 and Tyr36) in the beta-sheet side, might correspond to A-type blocking activity. A specific deletion mutant with the disrupted Face B, BmTx3-Y36P37del, loses the A-type current-blocking activity, but keeps a similar HERG-blocking activity, as seen with the wild-type toxin. PMID:14599291

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

    PubMed Central

    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

  6. Staphylococcus aureus of phage type 187 isolated from people occurred to be a genes carrier of eneterotoxin C and toxic shock syndrome toxin-1 (TSST-1).

    PubMed

    Piechowicz, Lidia; Garbacz, Katarzyna; Galiński, Janusz

    2008-07-01

    The aim of this study was to examine the genotype properties of Staphylococcus aureus of phage type 187 strains that constitute a separate group among the strains of S. aureus. Sixteen strains were collected from the hospital patients (n=12) and the healthy carriers (n=4) in 13 medical centres in Poland during 1991 and 2005. Biotyping, antibiotic susceptibility, phage typing, detection the genes of enterotoxins and toxic shock syndrome toxin, genotyping of chromosomal DNA by pulsed-field gel electrophoresis (PFGE), also amplification and restriction analysis of the coagulase (coa) and the protein A genes (spa) (PCR/restriction fragment length polymorphism (RFLP)) was tested. The results of this study showed that all staphylococcus of phage type 187 belonged to the human biotype (A) and appeared to be sensitive to all of the tested antibiotics, including methicillin (MSSA). Finding out the toxin genes showed that almost all of them (93.8%) had the enterotoxin C gene (sec) and TSST-1 gene (tst). The PFGE typing proved that the phage type 187 strains (except for one) constitute one PFGE type. These results and the identical restriction patterns in the PCR/RFLP method, also the same biotype, sensitivity to antibiotics and the presence genes of the same type of toxins confirmed that the phage type 187 strains constitute one clone within our country. Additionally, the fact that almost all of them have the enterotoxin genes and tst gene allows to consider them the strains of potentially high virulence.

  7. Graphene via Molecule-Assisted Ultrasound-Induced Liquid-Phase Exfoliation: A Supramolecular Approach

    NASA Astrophysics Data System (ADS)

    Eredia, Matilde; Ciesielski, Artur; Samorì, Paolo

    2016-12-01

    Graphene is a two-dimensional (2D) material holding unique optical, mechanical, thermal and electrical properties. The combination of these exceptional characteristics makes graphene an ideal model system for fundamental physical and chemical studies as well as technologically ground breaking material for a large range of applications. Graphene can be produced either following a bottom-up or top-down method. The former is based on the formation of covalent networks suitably engineered molecular building blocks undergoing chemical reaction. The latter takes place through the exfoliation of bulk graphite into individual graphene sheets. Among them, ultrasound-induced liquid-phase exfoliation (UILPE) is an appealing method, being very versatile and applicable to different environments and on various substrate types. In this chapter, we describe the recently reported methods to produce graphene via molecule-assisted UILPE of graphite, aiming at the generation of high-quality graphene. In particular, we will focus on the supramolecular approach, which consists in the use of suitably designed organic molecules during the UILPE of graphite. These molecules act as graphene dispersion-stabilizing agents during the exfoliation. This method relying on the joint effect of a solvent and ad hoc molecules to foster the exfoliation of graphite into graphene in liquid environment represents a promising and modular method toward the improvement of the process of UILPE in terms of the concentration and quality of the exfoliated material. Furthermore, exfoliations in aqueous and organic solutions are presented and discussed separately.

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

    PubMed Central

    De Boulle, Koenraad; Fagien, Steven; Sommer, Boris; Glogau, Richard

    2010-01-01

    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. PMID:20458348

  9. Ultrasound exfoliation of inorganic analogues of graphene

    NASA Astrophysics Data System (ADS)

    Štengl, Václav; Henych, Jiří; Slušná, Michaela; Ecorchard, Petra

    2014-04-01

    High-intensity ultrasound exfoliation of a bulk-layered material is an attractive route for large-scale preparation of monolayers. The monolayer slices could potentially be prepared with a high yield (up to 100%) in a few minutes. Exfoliation of natural minerals (such as tungstenite and molybdenite) or bulk synthetic materials (including hexagonal boron nitride (h-BN), hexagonal boron carbon nitride (h-BCN), and graphitic carbon nitride (g-C3N4)) in liquids leads to the breakdown of the 3D graphitic structure into a 2D structure; the efficiency of this process is highly dependent upon the physical effects of the ultrasound. Atomic force microscopy (AFM), transmission electron microscopy (TEM), and selected area electron diffraction (SAED) were employed to verify the quality of the exfoliation. Herein, this new method of exfoliation with ultrasound assistance for application to mono- and bilayered materials in hydrophobic and hydrophilic environments is presented.

  10. LYSOSOMAL DISRUPTION BY BACTERIAL TOXINS

    PubMed Central

    Bernheimer, Alan W.; Schwartz, Lois L.

    1964-01-01

    Bernheimer, Alan W. (New York University School of Medicine, New York), and Lois L. Schwartz. Lysosomal disruption by bacterial toxins. J. Bacteriol. 87:1100–1104. 1964.—Seventeen bacterial toxins were examined for capacity (i) to disrupt rabbit leukocyte lysosomes as indicated by decrease in turbidity of lysosomal suspensions, and (ii) to alter rabbit liver lysosomes as measured by release of β-glucuronidase and acid phosphatase. Staphylococcal α-toxin, Clostridium perfringens α-toxin, and streptolysins O and S affected lysosomes in both systems. Staphylococcal β-toxin, leucocidin and enterotoxin, Shiga neurotoxin, Serratia endotoxin, diphtheria toxin, tetanus neurotoxin, C. botulinum type A toxin, and C. perfringens ε-toxin were not active in either system. Staphylococcal δ-toxin, C. histolyticum collagenase, crude C. perfringens β-toxin, and crude anthrax toxin caused lysosomal damage in only one of the test systems. There is a substantial correlation between the hemolytic property of a toxin and its capacity to disrupt lysosomes, lending support to the concept that erythrocytes and lysosomes are bounded by similar membranes. PMID:5874534

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

    PubMed Central

    Intiso, Domenico; Basciani, Mario; Santamato, Andrea; Intiso, Marta; Di Rienzo, Filomena

    2015-01-01

    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 including post

  12. [Presence of enterotoxin C and toxic shock syndrome toxin--1 (TSST-1) genes in population of Staphylococcus aureus phage type 187].

    PubMed

    Garbacz, Katarzyna; Piechowicz, Lidia; Galiński, Janusz

    2006-01-01

    The aim of this study was to examine whether Staphylococcus aureus of phage type 187 possess the genes of enterotoxins and toxic shock syndrom toxin. Sixteen phage type 187 strains were isolated from the hospital patients (12) and the carriers (4) in twelve medical centres in Poland during 1991 and 2005. Biotyping, phage typing, antibiotic susceptibility, detection of the genes of enterotoxins (sea--sed) and toxic shock syndrome toxin (tst) was tested. The results of this study showed that all staphylococci of phage type 187 belonged to the human biotype (A) and appeared to be sensitive to all of the tested antibiotics, including methicillin (MSSA). Almost all of them (93.8%) had the enterotoxin C gene and TSST-1 gene. This fact allows to consider them the strains of potentially high virulence.

  13. Phenotypic H-Antigen Typing by Mass Spectrometry Combined with Genetic Typing of H Antigens, O Antigens, and Toxins by Whole-Genome Sequencing Enhances Identification of Escherichia coli Isolates

    PubMed Central

    Chui, Huixia; Domish, Larissa; Sloan, Angela; Hernandez, Drexler; McCorrister, Stuart; Robinson, Alyssia; Walker, Matthew; Peterson, Lorea A. M.; Majcher, Miles; Ratnam, Sam; Haldane, David J. M.; Bekal, Sadjia; Wylie, John; Chui, Linda; Tyler, Shaun; Xu, Bianli; Reimer, Aleisha; Nadon, Celine; Knox, J. David

    2016-01-01

    Mass spectrometry-based phenotypic H-antigen typing (MS-H) combined with whole-genome-sequencing-based genetic identification of H antigens, O antigens, and toxins (WGS-HOT) was used to type 60 clinical Escherichia coli isolates, 43 of which were previously identified as nonmotile, H type undetermined, or O rough by serotyping or having shown discordant MS-H and serotyping results. Whole-genome sequencing confirmed that MS-H was able to provide more accurate data regarding H antigen expression than serotyping. Further, enhanced and more confident O antigen identification resulted from gene cluster based typing in combination with conventional typing based on the gene pair comprising wzx and wzy and that comprising wzm and wzt. The O antigen was identified in 94.6% of the isolates when the two genetic O typing approaches (gene pair and gene cluster) were used in conjunction, in comparison to 78.6% when the gene pair database was used alone. In addition, 98.2% of the isolates showed the existence of genes for various toxins and/or virulence factors, among which verotoxins (Shiga toxin 1 and/or Shiga toxin 2) were 100% concordant with conventional PCR based testing results. With more applications of mass spectrometry and whole-genome sequencing in clinical microbiology laboratories, this combined phenotypic and genetic typing platform (MS-H plus WGS-HOT) should be ideal for pathogenic E. coli typing. PMID:27307455

  14. Botulinum Toxin Type A Targets RhoB to Inhibit Lysophosphatidic Acid-Stimulated Actin Reorganization and Acetylcholine Release in NGF-Treated Differentiated PC12 Cells

    DTIC Science & Technology

    2003-07-01

    from Sternberger Monoclonals (Lutherville, MD). Botulinum toxin type A (BoNT/A) was procured from Wako (Richmond, VA). 1 Report Documentation...Page Form ApprovedOMB No. 0704-0188 Public reporting burden for the collection of information is estimated to average 1 hour per response, including the...failing to comply with a collection of information if it does not display a currently valid OMB control number. 1 . REPORT DATE 01 JUL 2003 2. REPORT TYPE

  15. Botulinum toxin type A reduces TRPV1 expression in the dorsal root ganglion in rats with adjuvant-arthritis pain.

    PubMed

    Fan, Chenglei; Chu, Xiao; Wang, Lin; Shi, Hao; Li, Tieshan

    2017-07-01

    Arthritis pain affects people's long-term health, and recent studies have demonstrated that transient receptor potential vanilloid type 1 (TRPV1) plays a crucial role in arthritis pain. In addition, Pre-clinical evidence indicated that botulinum toxin type A (BoNT/A) has antinociceptive effect. The present study investigated the causality between the antinociceptive effects of BoNT/A and the expression of TRPV1 in dorsal root ganglion (DRG) in rats with adjuvant-arthritis pain. The results showed that BoNT/A significantly reduced adjuvant-arthritis nociceptive behaviors in a dose-dependent manner. Furthermore, the BoNT/A cleaved synaptosomal-associated protein of 25 kDa (cl-SNAP-25) was detected in the DRG using immunofluorescence after intra-articular administration. Although BoNT/A significantly reduced the protein levels of TRPV1, there were no significant changes in the mRNA levels of TRPV1 between CFA and BoNT/A (1U, 3U, 10U) group after BoNT/A retrograde axonal transport into the DRG with quantitative RT-PCR. This research provides evidence that the antinociceptive mechanism of BoNT/A might be mediated by reduction of TRPV1 expression through inhibition of its plasma membrane trafficking after intra-articular administration. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Aqueous Dispersions of Graphene from Electrochemically Exfoliated Graphite.

    PubMed

    Sevilla, Marta; Ferrero, Guillermo A; Fuertes, Antonio B

    2016-11-21

    A facile and environmentally friendly synthetic strategy for the production of stable and easily processable dispersions of graphene in water is presented. This strategy represents an alternative to classical chemical exfoliation methods (for example the Hummers method) that are more complex, harmful, and dangerous. The process is based on the electrochemical exfoliation of graphite and includes three simple steps: 1) the anodic exfoliation of graphite in (NH4 )2 SO4 , 2) sonication to separate the oxidized graphene sheets, and 3) reduction of oxidized graphene to graphene. The procedure makes it possible to convert around 30 wt % of the initial graphite into graphene with short processing times and high yields. The graphene sheets are well dispersed in water, have a carbon/oxygen atomic ratio of 11.7, a lateral size of about 0.5-1 μm, and contain only a few graphene layers, most of which are bilayer sheets. The processability of this type of aqueous dispersion has been demonstrated in the fabrication of macroscopic graphene structures, such as graphene aerogels and graphene films, which have been successfully employed as absorbents or as electrodes in supercapacitors, respectively. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. PREDICTION OF OXIDE SCALE EXFOLIATION IN STEAM TUBES

    SciTech Connect

    Sabau, Adrian S; Wright, Ian G

    2010-01-01

    Numerical simulation results are presented for the prediction of the likelihood of oxide scale exfoliation from superheater tubes. The scenarios considered involved alloys T22, TP347H, and TP347HFG subjected to a simplified operating cycle in a power plant generating supercritical steam. The states of stress and strain of the oxides grown in steam were based solely on modeling the various phenomena experienced by superheater tubes during boiler operation, current understanding of the oxidation behavior of each alloy in steam, and consideration of operating parameters such as heat flux, tube dimensions, and boiler duty cycle. Interpretation of the evolution of strain in these scales, and the approach to conditions where scale failure (hence exfoliation) is expected, makes use of the type of Exfoliation Diagrams that incorporate various cracking and exfoliation criteria appropriate for the system considered. In these diagrams, the strain accumulation with time in an oxide is represented by a strain trajectory derived from the net strain resulting from oxide growth, differences in coefficients of thermal expansion among the components, and relaxation due to creep. It was found that an oxide growing on a tube subjected to routine boiler load cycling conditions attained relatively low values of net strain, indicating that oxide failure would not be expected to occur during normal boiler operation. However, during a boiler shut-down event, strains sufficient to exceed the scale failure criteria were developed after times reasonably in accord with plant experience, with the scales on the ferritic steel failing in tension, and those on the austenitic steels in compression. The results presented illustrate that using this approach to track the state of strain in the oxide scale through all phases of boiler operation, including transitions from full-to-low load and shut-down events, offers the possibility of identifying the phase(s) of boiler operation during which oxide

  18. Enhanced expression of recombinant beta toxin of Clostridium perfringens type B using a commercially available Escherichia coli strain.

    PubMed

    Bakhshi, Fatemah; Pilehchian Langroudi, Reza; Imani, Bahram Golestani

    2016-06-30

    Clostridium perfringens beta toxin is only produced by types B and C and plays an important role in many human and animal diseases, causing fatal conditions that originate in the intestines. We compared the expression of C. perfringens type B vaccine strain recombinant beta toxin gene in the Escherichia coli strains RosettaTM(DE3) and BL21(DE3). The beta toxin gene was extracted from pJETβ and ligated with pET22b(+). pET22β was transformed into E. coli strains BL21(DE3) and RosettaTM(DE3). Recombinant protein was expressed as a soluble protein after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction in strain RosettaTM(DE3) but not in BL21(DE3). Expression was optimised by growing recombinant cells at 37 °C and at an induction of 0.5 mM, 1 mM, 1.5 mM IPTG. Expression was evaluated using sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE). The recombinant protein was purified via Ni-NTA and was analysed using western blot. We concluded that E. coli strain RosettaTM(DE3) can enhance the expression of C. perfringens recombinant beta toxin.

  19. Comparison of clinical marking and ultrasound-guided injection of Botulinum type A toxin into the masseter muscles for treating bruxism and its cosmetic effects.

    PubMed

    Quezada-Gaon, Natacha; Wortsman, Ximena; Peñaloza, Osvaldo; Carrasco, Juan Eduardo

    2016-09-01

    Botulinum toxin type A has been used for treating the hypertrophy of the masseter muscles and its cosmetic effects. Ultrasound is increasingly used in dermatology, along with the guidance of mini-invasive procedures. To evaluate the role of ultrasound for guiding the application of Botulinum A toxin in patients with cosmetic alterations due to bruxism, correlate the clinical landmarks with the ultrasound findings, and study the effect on the symptoms, cosmetics, and quality of life. Twenty individuals with bruxism and cosmetic alterations underwent an ultrasound-guided injection of Botulinum toxin type A in each masseter muscle. Clinical and ultrasound marking of the procedure was compared. Clinical and sonographic evaluation was performed at the time of injection and 3 months later. Ten normal individuals underwent ultrasound of the masseter muscles as a control group. Up to 65% of individuals showed anatomical variants of the salivary glands. The method for clinically marking the skin showed a frequently erroneous location of the anterior point (up to 40% of cases) that was proven by ultrasound to be out of the muscle. In 20% of cases, ultrasound showed that the needle should be longer to enter the muscle. After injection, most of the patients demonstrated a decrease of the symptoms and cosmetic and quality of life improvements. Ultrasound can be a potent tool for guiding the injection of Botulinum toxin into the masseter muscles. It may contribute to a more personalized procedure, better cosmetic results, and help to avoid potential complications. © 2016 Wiley Periodicals, Inc.

  20. Use of enzyme-linked immunoassays for antibody to types C and D botulinum toxins for investigations of botulism in cattle.

    PubMed

    Gregory, A R; Ellis, T M; Jubb, T F; Nickels, R J; Cousins, D V

    1996-02-01

    The development of specific enzyme-linked immunosorbent assays (ELISA) for antibody to types C and D Clostridium botulinum toxins for investigation of botulism in cattle is described. Partially purified type C and D toxins were used as antigens to develop these ELISAs. Specificity of the ELISAs was evaluated on sera from 333 adult beef and dairy cattle from areas with no history or evidence of botulism in animals or water birds. The test was also evaluated on sera from 41 herds that included herds vaccinated against botulism, confirmed botulism cases and herds from areas where the disease is considered endemic. The ELISAs detected the presence of antibody to botulinum toxins in samples from vaccinated cattle and both convalescent and clinically normal animals from unvaccinated herds with outbreaks of botulism. Antibody was also found in unvaccinated animals from herds in which there had been no diagnosed botulism cases in areas where botulism was considered endemic. Sera from some unvaccinated cattle with high ELISA reactivity was shown to be protective for mice in botulinum toxin neutralisation tests. The use of these tests in investigations of botulism in cattle is discussed.

  1. Botulinum toxin type A in post-stroke lower limb spasticity: a multicenter, double-blind, placebo-controlled trial.

    PubMed

    Kaji, Ryuji; Osako, Yuka; Suyama, Kazuaki; Maeda, Toshio; Uechi, Yasuyuki; Iwasaki, Masaru

    2010-08-01

    Lower limb spasticity in post-stroke patients can impair ambulation and reduces activities of daily living (ADL) performance of patients. Botulinum toxin type A (BoNTA) has been shown effective for upper limb spasticity. This study assesses the treatment of lower limb spasticity in a large placebo-controlled clinical trial. In this multicenter, randomized, double-blind, parallel-group, placebo-controlled study, we evaluate the efficacy and safety of one-time injections of botulinum toxin type A (BoNTA) in Japanese patients with post-stroke lower limb spasticity. One hundred twenty patients with lower limb spasticity were randomized to a single treatment with BoNTA 300 U or placebo. The tone of the ankle flexor was assessed at baseline and through 12 weeks using the Modified Ashworth Scale (MAS). Gait pattern and speed of gait were also assessed. The primary endpoint was area under the curve (AUC) of the change from baseline in the MAS ankle score. Significant improvement in spasticity with BoNTA 300 U was demonstrated by a mean difference in the AUC of the change from baseline in the MAS ankle score between the BoNTA and placebo groups (-3.428; 95% CIs, -5.841 to -1.016; p = 0.006; t test). A significantly greater decrease from baseline in the MAS ankle score was noted at weeks 4, 6 and 8 in the BoNTA group compared to the placebo group (p < 0.001). Significant improvement in the Clinicians Global Impression was noted by the investigator at weeks 4, 6 and 8 (p = 0.016-0.048, Wilcoxon test), but not by the patient or physical/occupational therapist. Assessments of gait pattern using the Physician's Rating Scale and speed of gait revealed no significant treatment differences but showed a tendency towards improvement with BoNTA. No marked difference was noted in the frequency of treatment-related adverse events between BoNTA and placebo groups. This was the first large-scale trial to indicate that BoNTA significantly reduced spasticity in lower limb muscles.

  2. Botulinum toxin type A in post-stroke lower limb spasticity: a multicenter, double-blind, placebo-controlled trial

    PubMed Central

    Osako, Yuka; Suyama, Kazuaki; Maeda, Toshio; Uechi, Yasuyuki; Iwasaki, Masaru

    2010-01-01

    Lower limb spasticity in post-stroke patients can impair ambulation and reduces activities of daily living (ADL) performance of patients. Botulinum toxin type A (BoNTA) has been shown effective for upper limb spasticity. This study assesses the treatment of lower limb spasticity in a large placebo-controlled clinical trial. In this multicenter, randomized, double-blind, parallel-group, placebo-controlled study, we evaluate the efficacy and safety of one-time injections of botulinum toxin type A (BoNTA) in Japanese patients with post-stroke lower limb spasticity. One hundred twenty patients with lower limb spasticity were randomized to a single treatment with BoNTA 300 U or placebo. The tone of the ankle flexor was assessed at baseline and through 12 weeks using the Modified Ashworth Scale (MAS). Gait pattern and speed of gait were also assessed. The primary endpoint was area under the curve (AUC) of the change from baseline in the MAS ankle score. Significant improvement in spasticity with BoNTA 300 U was demonstrated by a mean difference in the AUC of the change from baseline in the MAS ankle score between the BoNTA and placebo groups (−3.428; 95% CIs, −5.841 to −1.016; p = 0.006; t test). A significantly greater decrease from baseline in the MAS ankle score was noted at weeks 4, 6 and 8 in the BoNTA group compared to the placebo group (p < 0.001). Significant improvement in the Clinicians Global Impression was noted by the investigator at weeks 4, 6 and 8 (p = 0.016–0.048, Wilcoxon test), but not by the patient or physical/occupational therapist. Assessments of gait pattern using the Physician’s Rating Scale and speed of gait revealed no significant treatment differences but showed a tendency towards improvement with BoNTA. No marked difference was noted in the frequency of treatment-related adverse events between BoNTA and placebo groups. This was the first large-scale trial to indicate that BoNTA significantly reduced spasticity in lower limb

  3. RNA-based regulation in type I toxin-antitoxin systems and its implication for bacterial persistence.

    PubMed

    Berghoff, Bork A; Wagner, E Gerhart H

    2017-05-30

    Bacterial dormancy is a valuable survival strategy upon challenging environmental conditions. Dormant cells tolerate the consequences of high stress levels and may re-populate the environment upon return to favorable conditions. Antibiotic-tolerant bacteria-termed persisters-regularly cause relapsing infections, increase the likelihood of antibiotic resistance, and, therefore, earn increasing attention. Their generation often depends on toxins from chromosomal toxin-antitoxin systems. Here, we review recent insights concerning RNA-based control of toxin synthesis, and discuss possible implications for persister generation.

  4. Rock fall triggering from cyclic thermal forcing of exfoliation fractures

    NASA Astrophysics Data System (ADS)

    Collins, B. D.; Stock, G. M.

    2013-12-01

    Exfoliation of rocks cause cliffs to deteriorate through the formation and subsequent opening of fractures. Where human activities are exposed to this process, rock fall hazard is potentially high. A number of mechanisms (e.g., freeze-thaw, precipitation-induced seepage) are capable of triggering rock fall in steep rock masses, however, some rock falls occur in the absence of any identified trigger. Among the suggested causes for these unexplained rock falls is thermal heating which can lead to outward expansion of rock surfaces. Here we use data from three years of displacement transducer field monitoring and corroborated by lidar measurements to test this hypothesis by assessing the magnitude and temporal pattern of thermally-induced rock deformation in an exfoliating granitic landscape. We show that daily, seasonal, and annual temperature effects are sufficient to cause cyclic and cumulative deformation of fractures, and that fracture tip propagation with consequent detachment of rock masses likely results. We use thermodynamic and structural engineering principals to show that thermally-forced exfoliation surfaces (i.e., partially detached 'flakes' of rock) follow Carnot-cycle-type hysteresis loops (i.e., the cyclical conversion of thermal energy to work that occurs in a heat engine) and can be modeled by Euler-type buckling criteria (i.e., the resultant lateral deformation that occurs upon an increase in axial compressive loading, here, caused by thermal expansion along the axis of partially detached exfoliation flakes). Our data from a 19-m-tall, 4-m-wide, 10-cm-thick, instrumented granodiorite exfoliation flake indicate that cumulative annual outward deformation of up to 1 mm/yr occurs under fatigue crack growth conditions. These conditions are related to both cyclic thermal and mechanical forcing and gravitational loads acting along the surfaces that attach the flake to the rock mass. Based on these observations and our analyses, we suggest that the warmest

  5. Inhibition of cholera toxin and other AB toxins by polyphenolic compounds

    USDA-ARS?s Scientific Manuscript database

    All AB-type protein toxins have intracellular targets despite an initial extracellular location. These toxins use different methods to reach the cytosol and have different effects on the target cell. Broad-spectrum inhibitors against AB toxins are therefore hard to develop because the toxins use dif...

  6. Effectiveness of botulinum toxin type A treatment of neck pain related to nocturnal bruxism: a case report

    PubMed Central

    Santamato, Andrea; Panza, Francesco; Di Venere, Daniela; Solfrizzi, Vincenzo; Frisardi, Vincenza; Ranieri, Maurizio; Fiore, Pietro

    2010-01-01

    Objective This case report describes a patient with nocturnal bruxism and related neck pain treated with botulinum toxin type A (BTX-A). Clinical Features The patient was a 27-year-old man with nocturnal bruxism and difficulty in active mouth opening and chewing and neck pain at rest. His numeric pain score was 7 of 10. Surface electromyography of the temporalis and masseter muscles showed typical signs of hyperactivity, characterized by compound muscle action potential amplitude alterations. Intervention and Outcome After clinical evaluation, he was treated with BTX-A to reduce masseter and temporalis muscle hyperactivity. After 3 days of treatment with BTX-A, with each masseter muscle injected with a dose of about 40 mouse units with a dilution of 1 mL and with temporal muscle bilaterally injected with 25 mouse units with the same dilution, a decrease in bruxism symptoms was reported. Neck pain also decreased after the first treatment (visual analog scale of 2/10) and then resolved completely. After 4 weeks, electromyography showed the reduction of muscle hyperactivity with a decrease in the amplitude of the motor action potential. The same reduction in signs and symptoms was still present at assessment 3 months posttreatment. Conclusion These findings suggest that BTX-A may be a therapeutic option for the treatment of bruxism and related disorders. PMID:22027036

  7. Electrophysiological study for comparing the effect of biological activity between type A botulinum toxins in rat gastrocnemius muscle.

    PubMed

    Kim, C-S; Jang, W S; Son, I P; Nam, S H; Kim, Y I; Park, K Y; Kim, B J; Kim, M N

    2013-09-01

    New cosmetic applications and products based on the effects of botulinum toxin (BTX) treatment have stimulated demand for this class of natural compounds. This demand generates the need for appropriate standardized protocols to test and compare the effectiveness of new BTX preparations. Based on the previously described electrophysiological methods, we measured and compared the inhibitory effects of two BTX type A (BTX-A) preparations on neuromuscular transmission through split-body test. The effectiveness was evaluated in terms of the compound muscle action potential (CMAP) and conduction velocity after BTX-A injection. We used a split-body method to compare two different BTX-As in the rat. Based on the changes in the CMAP, the two different BTX-As induced paralytic effect on the rat tibialis anterior muscle. However, the two different BTX-A preparations did not differ significantly in effectiveness and did not induce a delay in conduction velocity. The new BTX-A preparation used in this electrophysiological study had similar effect compared with the previously marketed BTX-A.[AQ: Please approve the edits made to the sentence "The new BTX-A preparation…") We propose that a split-body electrophysiological protocol will be useful in establishing the comparative effectiveness of new BTX products.

  8. Inhibitory effect of botulinum toxin type A on the NANC system in rat respiratory models of neurogenic inflammation.

    PubMed

    Chien, Chiang-Ting; Lee, Hsin-Min; Wu, Chia-Ching Josh; Li, Ping-Chia

    2012-08-15

    This study investigated whether botulinum toxin type A (BTX-A) inhibits respiratory neurogenic inflammation in the non-adrenergic, non-cholinergic (NANC) transmitter system in rats. Neurogenic inflammation models were induced in Sprague Dawley (SD) rats through bilateral cerebral artery occlusion (BCAO) for different times (0, 30 and 60 min) or by stimulation with capsaicin at different doses (5 or 15 g/kg). Pre-Bötzinger Complex-Spikes and the expression of substance P, synaptosomal-associated protein-25 (SNAP-25), and reactive oxygen species (ROS) were detected with or without pretreatment of rats with BTX-A (15 or 30 U/kg). BCAO reduced pre-Bot C spike activity (spike/s) and increased the breath rate (breaths/s) in an unstable pattern in comparison to controls, while pretreatment with BTX-A slightly reduced this phenomenon. Pretreatment with BTX-A inhibited BCAO- or capsaicin-induced increases in expression of SNAP-25, substance P, and ROS in a dose-dependent manner in brainstem and lung tissue. BTX-A exerts a suppressive effect on neurogenic inflammation via non-adrenergic, non-cholinergic transmitters. These results add to the body of evidence elucidating the non-cholinergic effects of BTX-A in the context of neurogenic inflammation.

  9. Botulinum toxin type A injection for management of upper limb spasticity in children with cerebral palsy: a literature review.

    PubMed

    Park, Eun Sook; Rha, Dong-Wook

    2006-10-31

    The aim of this article was to present a review of the research literature on the outcome of botulinum toxin type A (BTX-A) injection for management of upper limb spasticity in children with cerebral palsy (CP). We searched the electronic databases of MEDLINE, CINAHL and PUBMED for all published studies with full-length English text available. For each study, the quality of the methods and the strength of evidence were assessed by 2 independent reviewers based on the American Academy for Cerebral Palsy and Developmental Medicine (AACPDM) guidelines. Four studies of level I, 8 studies of level IV and 4 studies of level V were identified. Due to the limited number of studies with high quality evidence and inconsistent results among studies, we were unable to support or refute the usefulness of BTX-A injection for management of upper limb spasticity in children with CP. Moreover, we identified several variables that may affect the outcome of injection, such as timing of age, dosage, dilution volumes, localization techniques of target muscles and participant characteristics. In summary, we have presented a review the literature and a discussion of the considerable uncertainty and variation associated with the clinical use of BTX-A injection for management of upper limb spasticity in children with CP.

  10. Botulinum Toxin Type A Injection for Management of Upper Limb Spasticity in Children with Cerebral Palsy: a Literature Review

    PubMed Central

    Rha, Dong-wook

    2006-01-01

    The aim of this article was to present a review of the research literature on the outcome of botulinum toxin type A (BTX-A) injection for management of upper limb spasticity in children with cerebral palsy (CP). We searched the electronic databases of MEDLINE, CINAHL and PUBMED for all published studies with full-length English text available. For each study, the quality of the methods and the strength of evidence were assessed by 2 independent reviewers based on the American Academy for Cerebral Palsy and Developmental Medicine (AACPDM) guidelines. Four studies of level I, 8 studies of level IV and 4 studies of level V were identified. Due to the limited number of studies with high quality evidence and inconsistent results among studies, we were unable to support or refute the usefulness of BTX-A injection for management of upper limb spasticity in children with CP. Moreover, we identified several variables that may affect the outcome of injection, such as timing of age, dosage, dilution volumes, localization techniques of target muscles and participant characteristics. In summary, we have presented a review the literature and a discussion of the considerable uncertainty and variation associated with the clinical use of BTX-A injection for management of upper limb spasticity in children with CP. PMID:17066503

  11. Estimation of botulinum toxin type A efficacy on spasticity and functional outcome in children with spastic cerebral palsy.

    PubMed

    Colovic, Hristina; Dimitrijevic, Lidija; Stankovic, Ivona; Nikolic, Dejan; Radovic-Janosevic, Dragana

    2012-03-01

    We evaluated the effects of botulinum toxin type A (BTA) - abobotulinumtoxinA on passive motion resistance (PMR) values of lower limbs affected muscles and on the functional motor status in children with spastic cerebral palsy (CP). In Group I (28 lower limbs with spastic muscles), and in Group II (14 lower limbs with dynamic spastic equinus) BTA was administered. Physical therapy was prescribed for 16 weeks. We estimated PMR using the Modified Ashworth Scale. Achieved functional motor level was evaluated by Gross Motor Function Classification System (GMFCS) and Gross Motor Function Measure (GMFM). Parameters were assessed before treatment and after 3,8,16 weeks and 6 months respectively. In Group I, PMR was significantly lower for hip adductors and knee extensors over 3-16 weeks, and for ankle joint extensors in both groups. There were significant differences for both groups in frequencies of GMFCS values after 16 weeks from BTA application. There was a significant increase in GMFM scores after 8 and 16 weeks from BTA application in both groups of patients. BTA treatment in CP children is followed by reduction in PMR values and improvement in functional motor status.

  12. Nanopore Sensing of Botulinum Toxin Type B by Discriminating an Enzymatically Cleaved Peptide from a Synaptic Protein Synaptobrevin 2 Derivative

    PubMed Central

    2015-01-01

    Botulinum neurotoxins (BoNTs) are the most lethal toxin known to human. Biodefense requires early and rapid detection of BoNTs. Traditionally, BoNTs can be detected by looking for signs of botulism in mice that receive an injection of human material, serum or stool. While the living animal assay remains the most sensitive approach, it is costly, slow and associated with legal and ethical constrains. Various biochemical, optical and mechanical methods have been developed for BoNTs detection with improved speed, but with lesser sensitivity. Here, we report a novel nanopore-based BoNT type B (BoNT-B) sensor that monitors the toxin’s enzymatic activity on its substrate, a recombinant synaptic protein synaptobrevin 2 derivative. By analyzing the modulation of the pore current caused by the specific BoNT-B-digested peptide as a marker, the presence of BoNT-B at a subnanomolar concentration was identified within minutes. The nanopore detector would fill the niche for a much needed rapid and highly sensitive detection of neurotoxins, and provide an excellent system to explore biophysical mechanisms for biopolymer transportation. PMID:25511125

  13. Side effects and potential risk factors of botulinum toxin type A intramuscular injections in knee flexion contractures of hemophiliacs.

    PubMed

    Rodriguez-Merchan, E Carlos; De la Corte-Rodriguez, Hortensia

    2017-07-01

    Knee flexion contracture (KFC) is a common complication of recurrent hemarthrosis in children and young adults with hemophilia. If the KFC is not prevented (by means of primary prophylaxis) and treated properly and early (be means of physical medicine and rehabilitation), it will become fixed. Areas covered: The aim of this article is to review the potential role of botulinum toxin type A (BTX-A) intramuscular injections for the treatment of KFC in people with hemophilia (PWH). Expert commentary: Although two recent reports have mentioned the benefits of intramuscular injections of BTX-A in PWH with KFC, the data are still scant and too preliminary. The use of intramuscular injections of BTX-A in PWH today should not be recommended until more case studies/small series (ideally well-designed clinical trials) fully demonstrate that this is safe and effective. The risks of intramuscular injections to a hemophilia patient cannot be underestimated (iatrogenic muscle hematomas and pseudotumors). This paper calls the attention of hemophilia treaters on the potential risks of this apparently interesting technique. The current use of BTX-A intramuscular injections in KFC of PWH could make no sense. Raising false expectations in these patients should be avoided.

  14. Comparing the Effect of Botulinum Toxin Type B Injection at Different Dosages for Patient with Drooling due to Brain Lesion

    PubMed Central

    Park, Hee Dong; Park, Sang Jun; Choi, Yong Min

    2012-01-01

    Objective To investigate Botulinum toxin type B (BNT-B) injection's effect and duration depending on dose for patients with brain lesion. Method Twenty one patients with brain lesion and severe drooling were included and divided into three groups. All patients received conventional dysphagia therapy. Group A patients (n=7) received an injection of 1,500 units and group B patients (n=7) received an injection of 2,500 units of BNT-B in submandibular gland under ultrasound guidance. Group C patients (n=7) received conventional dysphagia therapy. Saliva secretion was assessed quantitatively at baseline and at weeks 1, 2, 4, 8, and 12. The severity and frequency of drooling was assessed using the Drooling Quotient (DQ) by patients and/or caregivers. Results Group A and B reported a distinct improvement of the symptoms within 2 weeks after BNT-B injection. Compared to the baseline, the mean amount of saliva decreased significantly throughout the study. However, there was no meaningful difference between the two groups. The greatest reductions were achieved at 2 weeks and lasted up to 8 weeks after BNT-B injection. Group C did not show any differences. Conclusion Local injection of 1,500 units of BNT-B into salivary glands under ultrasonic guidance proved to be a safe and effective dose for drooling in patient with brain lesion, as did 2,500 units. PMID:23342318

  15. Persistence of Shiga toxin-producing Escherichia coli O26 in various manure-amended soil types.

    PubMed

    Fremaux, B; Prigent-Combaret, C; Delignette-Muller, M L; Mallen, B; Dothal, M; Gleizal, A; Vernozy-Rozand, C

    2008-01-01

    To evaluate the behaviour of Shiga toxin-producing Escherichia coli (STEC) O26 strains inoculated in manure-amended soils under in vitro conditions. Four green fluorescent protein (GFP)-labelled STEC O26 strains were inoculated in duplicate (at 10(6) CFU g(-1)) in three different manure-amended soil types, including two loam soils (A and B) and one clay loam soil (C), and two incubation temperatures (4 and 20 degrees C) were tested. STEC counts and soil physical parameters were periodically monitored. STEC O26 cells were able to persist during extended periods in soil even in the presence of low moisture levels, i.e. less than 0 x 08 g H2O g(-1) dry soil. At 4 and 20 degrees C, STEC could be detected in soil A for 288 and 196 days, respectively, and in soils B and C for at least 365 days postinoculation at both temperatures. The ambient temperature (i.e. 20 degrees C) was significantly associated with the highest STEC count decline in all soils tested. The temperature and soil properties appear to be contributory factors affecting the long-term survival of STEC O26 in manure-amended soils. This study provides useful information regarding the ecology of STEC O26 in manure-amended soils and may have implications for land and waste management.

  16. An Experimental Study on Botulinum Toxin Type A for the Treatment of Excessive Secretion after Submandibular Gland Transplantation in Rabbits

    PubMed Central

    Lin, Bo; Wang, Kan

    2016-01-01

    Objectives. To investigate whether botulinum toxin type A (BTXA) could control excessive secretion after submandibular gland (SMG) transplantation in rabbits and its possible mechanisms. Methods. A new SMG transplantation model was established in rabbit. 30 successfully constructed models were randomly assigned to five groups including control group and four experimental groups. Secretion outputs were used to analyze the effect of BTXA injection on excessive secretion. Hematoxylin and eosin (HE) staining, transmission electron microscopy (TEM), Western blot, and immunofluorescence were performed to analyze its possible mechanisms. Results. After BTXA injection, a significant decrease of excessive secretion after SMG transplantation was found in 2 and 4 weeks groups, but no significant effect on 12 and 24 weeks groups. HE and TEM results showed that BTXA led to morphological and ultrastructural changes of acinar cells of transplanted SMG. Western blot results suggested that BTXA decreased the aquaporin-5 (AQP5) protein expression after BTXA injection for 2 and 4 weeks. Immunofluorescence results showed that AQP5 protein was mainly expressed in the cytoplasm after BTXA injection for 2 and 4 weeks, which might indicate that BTXA promoted AQP5 expression from the cell membrane to cytoplasm. Conclusion. BTXA could effectively control excessive secretion after SMG transplantation in rabbits. PMID:27840738

  17. Botulinum toxin type A in simple motor tics: short-term and long-term treatment-effects.

    PubMed

    Rath, Judith J G; Tavy, Dénes L J; Wertenbroek, Agnes A A C M; van Woerkom, Theodoor C A M; de Bruijn, Sebastiaan F T M

    2010-08-01

    To determine the short-term and long-term treatment-effects of botulinum toxin type A in simple motor tics, we analyzed 15 consecutive patients (18 tics) with simple motor tics that were treated every 3 months with injections of BTX-A. Efficacy (rated on a 4-level scale) and duration of effect of the first 2 and last 2 (if treated 5 times or more) treatments were recorded, as well as latency of response, changes of premonitory urges (PMUs) and possible side effects. Total number of treatments for each tic varied from 2 to 50 (mean 11, median 6). In 16 of 18 tics (89%) short-term efficacy was reported successful (good or moderate). Long-term efficacy was reported in 12 tics of which 11 showed similar or even increased beneficial effects. Premonitory urge (PMU) was reported in 8 patients (53%). PMU, if present, lessened or disappeared after treatment with BTX-A. A permanent remission of the treated tic was seen in 3 patients with a maximum follow-up of 10 years. BTX-A appears a safe and effective treatment for simple motor tics and retains its efficacy after long-term treatment. BTX may also induce permanent remission of the treated tics and effects of BTX are not restricted to merely motor behaviour.

  18. Botulinum toxin type A combined with neurodynamic mobilization for upper limb spasticity after stroke: a case report

    PubMed Central

    Villafañe, Jorge H.; Silva, Guillermo B.; Chiarotto, Alessandro; Ragusa, Orazio L.F.

    2012-01-01

    Objective The purpose of this study is to report a case in which combinatory therapy of botulinum toxin type A (BoNT-A) and neurodynamic mobilization (NM) was used as treatment for a patient with severe upper limb spasticity and pain after stroke. Clinical Features A 76-year-old male patient had spastic muscles in the upper limb 10 months after an ischemic stroke. Intervention and Outcome The patient underwent combined treatment with BoNT-A and NM of the upper limb in 6 monthly applications. Evaluation was performed pretreatment, 3 months after the first injection, 3 months after the second injection, and at a follow-up session 9 months after starting the treatment. The following outcomes were measured: pain by using a numeric rating scale, spasticity by the Modified Ashworth Scale for Grading Spasticity, acceptance and emotional reaction to the treatment by the Hospital Anxiety and Depression Scale, and functionality by ranges of motion. The patient improved in all outcomes after treatment, and results were maintained during the follow-up sessions. Conclusion The combined NM and BoNT-A treatment appeared to decrease pain and improve joint ranges of motion during treatment for this patient. The patient showed decreased anxiety and depression during and after the treatment. PMID:23449315

  19. Efficacy and safety of botulinum toxin type A for treatment of Frey's syndrome: evidence from 22 published articles.

    PubMed

    Xie, Shang; Wang, Kan; Xu, Tao; Guo, Xue-Sheng; Shan, Xiao-Feng; Cai, Zhi-Gang

    2015-11-01

    Frey's syndrome (FS) is an unavoidable sequela following the surgery of the parotid gland. Although several treatment methods are available, their efficacy is short term or accompanied by unacceptable complications. In the past two decades, botulinum toxin type A (BTXA) has been widely used to treat FS. Although several systematic reviews have been reported recently, they were conflicting and with obvious deficiencies. Thus, we performed an objectively systematic review to determine whether BTXA is an effective and safe treatment for FS. A literature retrieval covering PubMed, Web of Science, Ovid, Embase and Cochrane library was performed on 16 January, 2015. Proportion meta-analysis and corresponding 95% confidence interval (CI) were performed to evaluate the efficacy and safety of BXTA in treatment of FS. A total of 499 records were retrieved and 22 articles with 23 studies were included after scrutiny by two independent authors. Statistical analyses regarding the effective rate, incidence of complications were used to estimate the efficacy and safety of BTXA. Our results suggested that the effective rate of BTXA for treatment of FS is 98.5% (95% CI = 0.971-0.994) and the incidence of complication is 3.6% (95% CI = 0.017-0.061). In conclusion, our study supports that BTXA produces meaningful benefits on the treatment of patients with FS. However, owing to lack of strong evidence, future studies with well-designed inclusion criteria and multicenter randomized controlled trials are needed to give more credible evidence, if possible.

  20. Vibrio type III effector VPA1380 is related to the cysteine protease domain of large bacterial toxins.

    PubMed

    Calder, Thomas; Kinch, Lisa N; Fernandez, Jessie; Salomon, Dor; Grishin, Nick V; Orth, Kim

    2014-01-01

    Vibrio parahaemolyticus is a Gram-negative halophilic bacterium and one of the leading causes of food-borne gastroenteritis. Its genome harbors two Type III Secretion Systems (T3SS1 and T3SS2), but only T3SS2 is required for enterotoxicity seen in animal models. Effector proteins secreted from T3SS2 have been previously shown to promote colonization of the intestinal epithelium, invasion of host cells, and destruction of the epithelial monolayer. In this study, we identify VPA1380, a T3SS2 effector protein that is toxic when expressed in yeast. Bioinformatic analyses revealed that VPA1380 is highly similar to the inositol hexakisphosphate (IP6)-inducible cysteine protease domains of several large bacterial toxins. Mutations in conserved catalytic residues and residues in the putative IP6-binding pocket abolished toxicity in yeast. Furthermore, VPA1380 was not toxic in IP6 deficient yeast cells. Therefore, our findings suggest that VPA1380 is a cysteine protease that requires IP6 as an activator.

  1. Treatment of upper limb spasticity after stroke: one-year safety and efficacy of botulinum toxin type A NT201.

    PubMed

    Fiore, P; Santamato, A; Ranieri, M; Bellomo, R G; Saggini, R; Panza, F; Megna, G; Cristella, G; Megna, M

    2012-01-01

    A new preparation of botulinum toxin type A called NT 201, free from complexing proteins, potentially with low antigenicity has been used in the therapy of spasticity in stroke patients. This was an open-label study reported the safety and the efficacy of one-year treatment with NT 201 evaluating the therapeutic effect on functional disability and on quality of life in upper limb spasticity after stroke. Patients received a botulinum toxin therapy in the upper injected intramuscularly. After inoculation, patients were submitted to a motor rehabilitation program for upper limb injected three times/week. Re-treatment was permitted at 12 weeks after the prior treatment. Safety assessment included evaluation of adverse events and efficacy was measured by Modified Ashworth Scale for spasticity (MAS), Spasm Frequency Score (SFS) for the daily spasms, and Disability Assessment Scale (DAS) for disability. Of 35 consecutive patients (13 women and 12 men) screened for study eligibility, 20 (6 women and 14 men) patients (mean age 63,4±7,03) were included in this study and were submitted to NT 201 therapy for one year. At the baseline, botulinum toxin dose in the upper limb ranged from 160 to 450U, whereas total dose in the last treatment administrated was reduced respect the first injections ranging from 120 to 350U. All the enrolled patients completed the year-long study and reported an improvement of clinical picture. MAS, was statistically (px003C;0,001) reduced in all muscles at T1 (mean score ±SD: 2.65±0.67) and T2 (mean score±SD: 2.55±0.60) in comparison to the baseline T0 (mean score±SD: 3.9 ±0.78). Significant reduction (px003C;0,001) from baseline T0 (mean score ±SD: 3.25±0.78) was also noted in SFS at T1 (mean score ±SD: 1.55±0.51) and T2 (mean score±SD :1.30±0.47). The DAS score showed a reduction of the T1score (mean score ±SD: 1.70±0.47) and T2 score (mean score ±SD: 1,40 ±0,50) respect to baseline T0 score (mean score ±SD: 2,65 ±0

  2. Infant botulism due to C. butyricum type E toxin: a novel environmental association with pet terrapins.

    PubMed

    Shelley, E B; O'Rourke, D; Grant, K; McArdle, E; Capra, L; Clarke, A; McNamara, E; Cunney, R; McKeown, P; Amar, C F L; Cosgrove, C; Fitzgerald, M; Harrington, P; Garvey, P; Grainger, F; Griffin, J; Lynch, B J; McGrane, G; Murphy, J; Ni Shuibhne, N; Prosser, J

    2015-02-01

    We describe two cases of infant botulism due to Clostridium butyricum producing botulinum type E neurotoxin (BoNT/E) and a previously unreported environmental source. The infants presented at age 11 days with poor feeding and lethargy, hypotonia, dilated pupils and absent reflexes. Faecal samples were positive for C. butyricum BoNT/E. The infants recovered after treatment including botulism immune globulin intravenous (BIG-IV). C. butyricum BoNT/E was isolated from water from tanks housing pet 'yellow-bellied' terrapins (Trachemys scripta scripta): in case A the terrapins were in the infant's home; in case B a relative fed the terrapin prior to holding and feeding the infant when both visited another relative. C. butyricum isolates from the infants and the respective terrapin tank waters were indistinguishable by molecular typing. Review of a case of C. butyricum BoNT/E botulism in the UK found that there was a pet terrapin where the infant was living. It is concluded that the C. butyricum-producing BoNT type E in these cases of infant botulism most likely originated from pet terrapins. These findings reinforce public health advice that reptiles, including terrapins, are not suitable pets for children aged <5 years, and highlight the importance of hand washing after handling these pets.

  3. Integron characterization and typing of Shiga toxin-producing Escherichia coli isolates in Belgium.

    PubMed

    Van Meervenne, Eva; Boon, Nico; Verstraete, Karen; Devlieghere, Frank; De Reu, Koen; Herman, Lieve; Buvens, Glenn; Piérard, Denis; Van Coillie, Els

    2013-05-01

    The presence of integrons and the antibiotic susceptibility profiles of STEC strains isolated in Belgium were analysed. The collection contained 306 strains, of which 225 were human isolates and 81 originated from different food or animal sources. Integrons were detected by PCR in 7.5% of the tested isolates and all were class 1 integrons. The integron-positive strains all belonged to the human collection. By RFLP, five different types (A, B, C, D, E) were distinguished. The antibiotic-resistance gene cassettes were identified by sequencing representatives of the five different types. Two types of gene cassettes were found in different combinations, one encoding resistance to streptomycin/spectinomycin and the other encoding resistance to trimethoprim. One of the gene cassettes present was the rarely detected aadA23, which was now apparently for the first time reported in Western Europe. Susceptibility profiling of the strains for 11 antibiotics was done by standard disc diffusion assays. Among the 23 integron-positive strains, 17 different antibiotic susceptibility profiles were found. In the 283 integron-negative strains, 24 different antibiotic susceptibility profiles were observed. The majority of these strains were susceptible to all tested antibiotics (n=218, 77.0%). The integron-positive strains were significantly more resistant to eight of the eleven tested antibiotics compared to the integron-negative strains (P<0.05). PFGE profiles of integron-positive strains within selected serogroups did not cluster together.

  4. Ultrasound-Guided Botulinum Toxin Type A Salivary Gland Injection in Children for Refractory Sialorrhea: 10-Year Experience at a Large Tertiary Children's Hospital.

    PubMed

    Lungren, Matthew P; Halula, Sarah; Coyne, Sarah; Sidell, Douglas; Racadio, John M; Patel, Manish N

    2016-01-01

    Sialorrhea is problematic for neurologically impaired children, and botulinum toxin A salivary gland injection has been reported as effective in reducing sialorrhea. This article assesses the success and safety of ultrasound-guided weight-based botulinum toxin A injection for the management of sialorrhea in children. A total of 111 patients (63 males; 48 females; average age 7 years) with refractory sialorrhea were treated with ultrasound-guided botulinum toxin type A salivary gland injections (144 procedures) from July 1, 2004, to July 1, 2014, using a single weight-based protocol. Patient history, procedural records, and clinical follow-up documents were retrospectively reviewed. Clinical data were compared with reported effectiveness and complications using odds ratios. A total of 144 procedures were performed in 111 patients with refractory sialorrhea. Cerebral palsy was the most common underlying etiology for sialorrhea (29%), whereas others included encephalopathy (5%), anoxic brain injury (4%), and a variety of chromosomal anomalies (5%). There was a 100% technical success rate. Overall treatment effectiveness was 68%. Repeat injections were not associated with increased clinical success. No procedure-related deaths or major complications were identified; the minor complication rate was less than 2%. The protocol used for ultrasound-guided injection of botulinum toxin A proved to be safe and effective in children suffering from sialorrhea. Image guidance technique may lead to a reduction in rates of adverse events reported in other series. Subsequent procedures do not improve upon initial efficacy. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. A vapBC-type toxin-antitoxin module of Sinorhizobium meliloti influences symbiotic efficiency and nodule senescence of Medicago sativa.

    PubMed

    Lipuma, Justine; Cinege, Gyöngyi; Bodogai, Monica; Oláh, Boglárka; Kiers, Aurélie; Endre, Gabriella; Dupont, Laurence; Dusha, Ilona

    2014-12-01

    The symbiotic nitrogen-fixing soil bacterium Sinorhizobium meliloti carries a large number of toxin-antitoxin (TA) modules both on the chromosome and megaplasmids. One of them, the vapBC-5 module that belongs to the type II systems was characterized here. It encodes an active toxin vapC-5, and was shown to be controlled negatively by the complex of its own proteins. Different mutants of the vapBC-5 genes exhibited diverse effects on symbiotic efficiency during interaction with the host plant Medicago sativa. The absence of the entire vapBC-5 region had no influence on nodule formation and nitrogen fixation properties. The strain carrying an insertion in the antitoxin gene showed a reduced nitrogen fixation capacity resulting in a lower plant yield. In contrast, when the toxin gene was mutated, the strain developed more efficient symbiosis with the host plant. The nitrogen fixing root nodules had a delayed senescent phenotype and contained elevated level of plant-derived molecules characteristic of later steps of nodule development. The longer bacteroid viability and abundance of active nitrogen fixing zone resulted in increased production of plant material. These data indicate that modification of the toxin/antitoxin production may influence bacteroid metabolism and may have an impact on the adaptation to changing environmental conditions.

  6. Mass Spectrometry-Based Method of Detecting and Distinguishing Type 1 and Type 2 Shiga-Like Toxins in Human Serum

    PubMed Central

    Silva, Christopher J.; Erickson-Beltran, Melissa L.; Skinner, Craig B.; Patfield, Stephanie A.; He, Xiaohua

    2015-01-01

    Shiga-like toxins (verotoxins) are responsible for the virulence associated with a variety of foodborne bacterial pathogens. Direct detection of toxins requires a specific and sensitive technique. In this study, we describe a mass spectrometry-based method of analyzing the tryptic decapeptides derived from the non-toxic B subunits. A gene encoding a single protein that yields a set of relevant peptides upon digestion with trypsin was designed. The 15N-labeled protein was prepared by growing the expressing bacteria in minimal medium supplemented with 15NH4Cl. Trypsin digestion of the 15N-labeled protein yields a set of 15N-labeled peptides for use as internal standards to identify and quantify Shiga or Shiga-like toxins. We determined that this approach can be used to detect, quantify and distinguish among the known Shiga toxins (Stx) and Shiga-like toxins (Stx1 and Stx2) in the low attomole range (per injection) in complex media, including human serum. Furthermore, Stx1a could be detected and distinguished from the newly identified Stx1e in complex media. As new Shiga-like toxins are identified, this approach can be readily modified to detect them. Since intact toxins are digested with trypsin prior to analysis, the handling of intact Shiga toxins is minimized. The analysis can be accomplished within 5 h. PMID:26633510

  7. Liquid exfoliation of defect-free graphene.

    PubMed

    Coleman, Jonathan N

    2013-01-15

    Due to its unprecedented physical properties, graphene has generated huge interest over the last 7 years. Graphene is generally fabricated in one of two ways: as very high quality sheets produced in limited quantities by micromechanical cleavage or vapor growth or as a rather defective, graphene-like material, graphene oxide, produced in large quantities. However, a growing number of applications would profit from the availability of a method to produce high-quality graphene in large quantities. This Account describes recent work to develop such a processing route inspired by previous theoretical and experimental studies on the solvent dispersion of carbon nanotubes. That work had shown that nanotubes could be effectively dispersed in solvents whose surface energy matched that of the nanotubes. We describe the application of the same approach to the exfoliation of graphite to give graphene in a range of solvents. When graphite powder is exposed to ultrasonication in the presence of a suitable solvent, the powder fragments into nanosheets, which are stabilized against aggregation by the solvent. The enthalpy of mixing is minimized for solvents with surface energies close to that of graphene (∼68 mJ/m(2)). The exfoliated nanosheets are free of defects and oxides and can be produced in large quantities. Once solvent exfoliation is possible, the process can be optimized and the nanosheets can be separated by size. The use of surfactants can also stabilize exfoliated graphene in water, where the ζ potential of the surfactant-coated graphene nanosheets controls the dispersed concentration. Liquid exfoliated graphene can be used for a range of applications: graphene dispersions as optical limiters, films of graphene flakes as transparent conductors or sensors, and exfoliated graphene as a mechanical reinforcement for polymer-based composites. Finally, we have extended this process to exfoliate other layered compounds such as BN and MoS(2). Such materials will be

  8. Synthesis and characterization of exfoliated graphene oxide

    NASA Astrophysics Data System (ADS)

    Muhamad, Ku Sarah Syahidah Ku; Mohamed, Faizal; Radiman, Shahidan; Hamzah, Ainon; Sarmani, Sukiman; Siong, Khoo Kok; Yasir, Muhammad Samudi; Rahman, Irman Abdul; Rosli, Nur Ratasha Alia Md.

    2016-11-01

    Graphene oxide has many applications such as in electronic devices, as storage energy device, biosensor, biomedical application, water purification, coating technology, as a composite and paper like materials. Hummer's method is one of the most common methods used in synthesizing graphene oxide. Graphene is different in size and structure because of oxidized layered of graphene oxide hence, the expanded interlayer structure of graphene oxide can be easily exfoliated by ultrasonication. We report on the preparation of exfoliated graphene oxide by using sonication method. Ultraviolet-visible spectrometer (UV-Vis) and Fourier-Transform Infrared Spectra Analyzer (FTIR) were used to characterize the exfoliated graphite oxide. Exfoliation of graphite oxide is conducted using water bath sonication. In order to confirm the chemical conformation and structure of the produced graphene oxide, FTIR and UV-Vis spectroscopy were utilized. Both peak of C=O and C-C bond are detected using UV-Vis and the results were confirmed using FTIR. Therefore, from this study, it can be concluded based on FTIR and UV-Vis spectral acquisition that graphene oxide can be produced by exfoliation of graphite oxide using water bath sonication.

  9. Anterior chamber angle in the exfoliation syndrome.

    PubMed Central

    Wishart, P K; Spaeth, G L; Poryzees, E M

    1985-01-01

    The gonioscopic findings of 76 patients with the exfoliation syndrome were reviewed. A high frequency of narrowness of the anterior chamber (AC) angle was found (32%). 18% had angles considered occludable, and 14% had obvious angle-closure glaucoma as shown by the presence of peripheral anterior synechias (PAS). Increased pigmentation of the posterior trabecular meshwork (PTM) was noted in all cases. When this pigmentation was markedly asymmetrical, unilateral exfoliation with glaucoma was common in the more pigmented eye. In addition heavy angle pigmentation in the absence of exfoliation was noted in the fellow eye of patients with characteristic exfoliated material in the other eye. Increased pigmentation of the PTM may be the earliest detectable sign of the exfoliation syndrome (ES). The clinical significance of our estimating PTM pigmentation at the 12 o'clock position is discussed. In view of the accelerated optic nerve damage associated with the development of glaucoma secondary to ES, routine estimation of the pigmentation of the PTM at 12 o'clock is recommended in the hope of early detection of cases of otherwise inapparent ES. Images PMID:3966996

  10. Crystal structure of Cry51Aa1: A potential novel insecticidal aerolysin-type β-pore-forming toxin from Bacillus thuringiensis.

    PubMed

    Xu, Chengchen; Chinte, Unmesh; Chen, Lirong; Yao, Qingqing; Meng, Ying; Zhou, Dayong; Bi, Li-Jun; Rose, John; Adang, Michael J; Wang, Bi-Cheng; Yu, Ziniu; Sun, Ming

    2015-07-03

    The structures of several Bacillus thuringiensis (Bt) insecticidal crystal proteins have been determined by crystallographic methods and a close relationship has been explicated between specific toxicities and conserved three-dimensional architectures. In this study, as a representative of the coleopteran- and hemipteran-specific Cry51A group, the complete structure of Cry51Aa1 protoxin has been determined by X-ray crystallography at 1.65 Å resolution. This is the first report of a coleopteran-active Bt insecticidal toxin with high structural similarity to the aerolysin-type β-pore forming toxins (β-PFTs). Moreover, study of featured residues and structural elements reveal their possible roles in receptor binding and pore formation events. This study provides new insights into the action of aerolysin-type β-PFTs from a structural perspective, and could be useful for the control of coleopteran and hemipteran insect pests in agricultures.

  11. Treatment of vital and non-vital primary molar teeth by one-stage formocresol pulpotomy: clinical success and effect upon age at exfoliation.

    PubMed

    Roberts, J F

    1996-06-01

    The clinical success and effect upon the age at which teeth exfoliated was prospectively observed in 175 primary molars that had received formoceresol pulpotomies performed by one operator. The success rate among 142 vital teeth was 99.3% and among 33 non-vital teeth 84.8%. There was no significant effect upon age at exfoliation after either type of pulpal treatment.

  12. Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro.

    PubMed Central

    Riegler, M; Sedivy, R; Pothoulakis, C; Hamilton, G; Zacherl, J; Bischof, G; Cosentini, E; Feil, W; Schiessel, R; LaMont, J T

    1995-01-01

    Toxin A but not toxin B, appears to mediate intestinal damage in animal models of Clostridium difficile enteritis. The purpose of this study was to investigate the electrophysiologic and morphologic effects of purified C. difficile toxins A and B on human colonic mucosa in Ussing chambers. Luminal exposure of tissues to 16-65 nM of toxin A and 0.2-29 nM of toxin B for 5 h caused dose-dependent epithelial damage. Potential difference, short-circuit current and resistance decreased by 76, 58, and 46%, respectively, with 32 nM of toxin A and by 76, 55, and 47%, respectively, with 3 nM of toxin B, when compared with baseline (P < 0.05). 3 nM of toxin A did not cause electrophysiologic changes. Permeability to [3H]mannitol increased 16-fold after exposure to 32 nM of toxin A and to 3 nM of toxin B when compared with controls (P < 0.05). Light and scanning electron microscopy after exposure to either toxin revealed patchy damage and exfoliation of superficial epithelial cells, while crypt epithelium remained intact. Fluorescent microscopy of phalloidin-stained sections showed that both toxins caused disruption and condensation of cellular F-actin. Our results demonstrate that the human colon is approximately 10 times more sensitive to the damaging effects of toxin B than toxin A, suggesting that toxin B may be more important than toxin A in the pathogenesis of C. difficile colitis in man. Images PMID:7738167

  13. Safety of botulinum toxin type A among children with spasticity secondary to cerebral palsy: a systematic review of randomized clinical trials.

    PubMed

    Albavera-Hernández, Cidronio; Rodríguez, Jorge M; Idrovo, Alvaro J

    2009-05-01

    To assess the safety of botulinum toxin type A for children with cerebral palsy using data from randomized clinical trials. A search was conducted of MEDLINE database for the period January 1990 to February 2008. A complementary search was carried out of references from previous reviews. From 122 articles, 20 randomized clinical trials were selected for the review. Two authors independently assessed eligibility of the studies and methodological quality of those according to CONSORT guidelines. For each adverse effect, relative risk was calculated, and when more than one study reported adverse effects, a meta-analysis was carried out. Sensitivity analyses with different continuity corrections factors were performed when zero cases were reported in a study arm. There were 882 participants in the 20 studies included in the meta-analysis. Six studies reported zero adverse effects. Thirty-five different adverse events were reported. Botulinum toxin type A use was related to respiratory tract infection, bronchitis, pharyngytis, asthma, muscle weakness, urinary incontinence, falls, seizures, fever and unspecified pain. Two deaths were reported in the study with higher person-time follow-up (relative risk (RR) 4.69, 95% confidence interval (CI) 0.23-95). Botulinum toxin type A has a good safety profile during the first months of use. However the occurrence of adverse events is more frequent among children with cerebral palsy than individuals with other conditions. Severe adverse events are potentially related with the use of botulinum toxin type A, but data are sparse and additional study is required to clarify the causal relation.

  14. MANAGING OXIDE SCALE EXFOLIATION IN BOILERS WITH TP347H SUPERHEATER TUBES

    SciTech Connect

    Sabau, Adrian S; Wright, Ian G.; Shingledecker, John P.; Tortorelli, Peter F

    2014-01-01

    A model based on a concept of fraction of exfoliated area as a function of oxide scale strain energy was developed to predict the extent of exfoliation of steam-side scale from boiler tube superheater loops. As compared with the Armitt diagram, which can be used to predict when scale damage and exfoliation would be likely to occur, a fraction of exfoliated area approach provides an estimation of mass of scale released and the fraction of tube likely to be blocked by the exfoliation. This paper show results for the extent of blockage expected in a single bend of a superheater loop was predicted as a function of operating time, bend geometry, and outlet steam temperature under realistic service conditions that include outages. The deposits of exfoliated scale were assumed to be distributed horizontally the tubes bends. Three types of bends were considered: regular bends, short bends, and hairpin bends. The progressive increase in steam and tube temperatures along a single loop of superheater tubing and the ensuing variation of oxide scale thickness are considered. Numerical simulation results for a superheater loop made of TP347H austenitic steel indicated that tube blockage fractions larger than 50% are likely to occur within the first two years of boiler operation (with regularly scheduled outages) for outlet tube temperatures of 540-570oC, which is consistent with practical experience. Higher blockage fractions were predicted for tubes with short bends and hairpin bends than for tubes with regular bends, of length that are larger than five internal tube diameters. Finally, the blockage model presented can be used with some confidence to devise operating schedules for managing the consequences of oxide scale exfoliation based on projections of time to some critical blockage fraction for specific boiler operating conditions.

  15. Differential Response of the Human Renal Proximal Tubular Epithelial Cell Line HK-2 to Shiga Toxin Types 1 and 2 ▿

    PubMed Central

    Lentz, Erin K.; Leyva-Illades, Dinorah; Lee, Moo-Seung; Cherla, Rama P.; Tesh, Vernon L.

    2011-01-01

    Shiga toxins (Stxs) are expressed by the enteric pathogens Shigella dysenteriae serotype 1 and certain serotypes of Escherichia coli. Stx-producing bacteria cause bloody diarrhea with the potential to progress to acute renal failure. Stxs are potent protein synthesis inhibitors and are the primary virulence factors responsible for renal damage that may follow diarrheal disease. We explored the use of the immortalized human proximal tubule epithelial cell line HK-2 as an in vitro model of Stx-induced renal damage. We showed that these cells express abundant membrane Gb3 and are differentially susceptible to the cytotoxic action of Stxs, being more sensitive to Shiga toxin type 1 (Stx1) than to Stx2. At early time points (24 h), HK-2 cells were significantly more sensitive to Stxs than Vero cells; however, by 72 h, Vero cell monolayers were completely destroyed while some HK-2 cells survived toxin challenge, suggesting that a subpopulation of HK-2 cells are relatively toxin resistant. Fluorescently labeled Stx1 B subunits localized to both lysosomal and endoplasmic reticulum (ER) compartments in HK-2 cells, suggesting that differences in intracellular trafficking may play a role in susceptibility to Stx-mediated cytotoxicity. Although proinflammatory cytokines were not upregulated by toxin challenge, Stx2 selectively induced the expression of two chemokines, macrophage inflammatory protein-1α (MIP-1α) and MIP-1β. Stx1 and Stx2 differentially activated components of the ER stress response in HK-2 cells. Finally, we demonstrated significant poly(ADP-ribose) polymerase (PARP) cleavage after exposure to Stx1 or Stx2. However, procaspase 3 cleavage was undetectable, suggesting that HK-2 cells may undergo apoptosis in response to Stxs in a caspase 3-independent manner. PMID:21708996

  16. Pertussis toxin

    SciTech Connect

    Sekura, R.D.; Moss, J.; Vaughan, M.

    1985-01-01

    This book contains 13 selections. Some of the titles are: Genetic and Functional Studies of Pertussis Toxin Substrates; Effect of Pertussis Toxin on the Hormonal Responsiveness of Different Tissues; Extracellular Adenylate Cyclase of Bordetella pertussis; and GTP-Regulatory Proteins are Introcellular Messagers: A Model for Hormone Action.

  17. Effectiveness of switching therapy from complexing protein-containing botulinum toxin type A to a formulation with low immunogenicity in spasticity after stroke: a case report.

    PubMed

    Santamato, Andrea; Ranieri, Maurizio; Panza, Francesco; Frisardi, Vincenza; Micello, Maria Francesca; Filoni, Serena; Fiore, Pietro

    2012-09-01

    Some patients receiving botulinum toxin type A therapy develop immunological resistance due to the production of neutralizing antibodies against the neurotoxin, thus partially or completely reducing the therapeutic effect. We report here neurophysiological and clinical findings for a 58-year-old man treated with botulinum toxin type A for spasticity after ischaemic stroke, who became a secondary non-responder patient. Subsequent treatment with a different preparation of botulinum toxin type A had a great therapeutic effect on his spasticity. The muscles injected and the dosages were the same for each treatment, but evaluation with the Modified Ashworth Scale after treatment with the second preparation showed a reduction of approximately 2 points compared with the first examination. The clinical results were also supported by extensor digitorum brevis testing of the right muscle, which showed a reduction in compound muscle action potential, whereas it was unchanged in the non-injected muscle. No side-effects were reported, and after 1 year of treatment with this formulation clinical benefits were still evident. The neurophysiological and clinical results obtained in this patient suggest that switching therapy from a complexing protein-containing product to a product potentially free of complexing proteins, which has low immunogenicity, may be a viable therapeutic option in secondary non-responder patients.

  18. Effects of intra-articular botulinum toxin type A (Botox) in dogs with chronic osteoarthritis.

    PubMed

    Hadley, H S; Wheeler, J L; Petersen, S W

    2010-01-01

    To evaluate the effects of intra-articular botulinum neurotoxin type A (BoNT/A) in dogs with chronic osteo- arthritis. Client-owned dogs with lameness and discomfort attributed to unilateral elbow or hip osteoarthritis were eligible for inclusion (n = 5). All dogs had BoNT/A (25 units) administered to the affected joint (2 elbows, 3 hips). Dogs were evaluated by pressure platform gait analysis before and at two, four, eight, and 12 weeks post-injection, and by client perception of outcome. In experimental limbs, ground reaction forces (peak vertical force and vertical impulse) consistently improved for a variable period of time following intra-articular BoNT/A therapy. These changes were not, however, observed in the contralateral limbs, in which values remained relatively unchanged or decreased. Four out of five owners reported at least some improvement in their dog's condition following treatment. A multimodal approach with the intra-articular administration of BoNT/A may be an option for osteoarthritis patients that are unresponsive to medical management and unable to undergo surgery. However, the findings of this study are preliminary and must be verified by further investigation.

  19. Chemically exfoliated ReS2 nanosheets.

    PubMed

    Fujita, Takeshi; Ito, Yoshikazu; Tan, Yongwen; Yamaguchi, Hisato; Hojo, Daisuke; Hirata, Akihiko; Voiry, Damien; Chhowalla, Manish; Chen, Mingwei

    2014-11-07

    The production of two-dimensional rhenium disulfide (ReS2) nanosheets by exfoliation using lithium intercalation is demonstrated. The vibrational and photoluminescence properties of the exfoliated nanosheets are investigated, and the local atomic structure is studied by scanning and transmission electron microscopy. The catalytic activity of the nanosheets in a hydrogen evolution reaction (HER) is also investigated. The electrochemical properties of the exfoliated ReS2 nanosheets include low overpotentials of ∼100 mV and low Tafel slopes of 75 mV dec(-1) for HER and are attributed to the atomic structure of the superlattice 1T' phase. The presence of bandgap photoluminescence demonstrates that the nanosheets retain their semiconducting nature. ReS2 nanosheets produced by this method provide unique photocatalytic properties that are superior to those of other two-dimensional systems.

  20. Immunization with a chimera consisting of the B subunit of Shiga toxin type 2 and brucella lumazine synthase confers total protection against Shiga toxins in mice.

    PubMed

    Mejias, María P; Ghersi, Giselle; Craig, Patricio O; Panek, Cecilia A; Bentancor, Leticia V; Baschkier, Ariela; Goldbaum, Fernando A; Zylberman, Vanesa; Palermo, Marina S

    2013-09-01

    The striking feature of enterohemorrhagic Escherichia coli (EHEC) infections is the production of Shiga toxins (Stx) implicated in the development of the life-threatening hemolytic uremic syndrome. Despite the magnitude of the social impact of EHEC infections, no licensed vaccine or effective therapy is available for human use. One of the biggest challenges is to develop an effective and safe immunogen to ensure nontoxicity, as well as a strong input to the immune system to induce long-lasting, high-affinity Abs with anti-Stx-neutralizing capacity. The enzyme lumazine synthase from Brucella spp. (BLS) is a highly stable dimer of pentamers and a scaffold with enormous plasticity on which to display foreign Ags. Taking into account the advantages of BLS and the potential capacity of the B subunit of Stx2 to induce Abs that prevent Stx2 toxicity by blocking its entrance into the host cells, we engineered a new immunogen by inserting the B subunit of Stx2 at the amino termini of BLS. The resulting chimera demonstrated a strong capacity to induce a long-lasting humoral immune response in mice. The chimera induced Abs with high neutralizing capacity for Stx2 and its variants. Moreover, immunized mice were completely protected against i.v. Stx2 challenge, and weaned mice receiving an oral challenge with EHEC were completely protected by the transference of immune sera. We conclude that this novel immunogen represents a promising candidate for vaccine or Ab development with preventive or therapeutic ends, for use in hemolytic uremic syndrome-endemic areas or during future outbreaks caused by pathogenic strains of Stx-producing E. coli.

  1. Chemically exfoliated ReS2 nanosheets

    NASA Astrophysics Data System (ADS)

    Fujita, Takeshi; Ito, Yoshikazu; Tan, Yongwen; Yamaguchi, Hisato; Hojo, Daisuke; Hirata, Akihiko; Voiry, Damien; Chhowalla, Manish; Chen, Mingwei

    2014-10-01

    The production of two-dimensional rhenium disulfide (ReS2) nanosheets by exfoliation using lithium intercalation is demonstrated. The vibrational and photoluminescence properties of the exfoliated nanosheets are investigated, and the local atomic structure is studied by scanning and transmission electron microscopy. The catalytic activity of the nanosheets in a hydrogen evolution reaction (HER) is also investigated. The electrochemical properties of the exfoliated ReS2 nanosheets include low overpotentials of ~100 mV and low Tafel slopes of 75 mV dec-1 for HER and are attributed to the atomic structure of the superlattice 1T' phase. The presence of bandgap photoluminescence demonstrates that the nanosheets retain their semiconducting nature. ReS2 nanosheets produced by this method provide unique photocatalytic properties that are superior to those of other two-dimensional systems.The production of two-dimensional rhenium disulfide (ReS2) nanosheets by exfoliation using lithium intercalation is demonstrated. The vibrational and photoluminescence properties of the exfoliated nanosheets are investigated, and the local atomic structure is studied by scanning and transmission electron microscopy. The catalytic activity of the nanosheets in a hydrogen evolution reaction (HER) is also investigated. The electrochemical properties of the exfoliated ReS2 nanosheets include low overpotentials of ~100 mV and low Tafel slopes of 75 mV dec-1 for HER and are attributed to the atomic structure of the superlattice 1T' phase. The presence of bandgap photoluminescence demonstrates that the nanosheets retain their semiconducting nature. ReS2 nanosheets produced by this method provide unique photocatalytic properties that are superior to those of other two-dimensional systems. Electronic supplementary information (ESI) available: Experimental procedures for sample synthesis and characterization. See DOI: 10.1039/c4nr03740e

  2. Toward the identification of a type I toxin-antitoxin system in the plasmid DNA of dairy Lactobacillus rhamnosus.

    PubMed

    Folli, Claudia; Levante, Alessia; Percudani, Riccardo; Amidani, Davide; Bottazzi, Stefania; Ferrari, Alberto; Rivetti, Claudio; Neviani, Erasmo; Lazzi, Camilla

    2017-09-21

    Plasmids carry genes that give bacteria beneficial traits and allow them to survive in competitive environments. In many cases, they also harbor toxin-antitoxin (TA) systems necessary for plasmid maintenance. TA systems are generally characterized by a stable "toxin", a protein or peptide capable of killing the cell upon plasmid loss and by an unstable "antitoxin", a protein or a non-coding RNA that inhibits toxin activity. Here we report data toward the identification of a RNA-regulated TA system in the plasmid DNA of L. rhamnosus isolated from cheese. The proposed TA system comprises two convergently transcribed RNAs: a toxin RNA encoding a 29 amino acid peptide named Lpt and an antitoxin non-coding RNA. Both toxin and antitoxin RNAs resulted upregulated under conditions mimicking cheese ripening. The toxicity of the Lpt peptide was demonstrated in E. coli by cloning the Lpt ORF under the control of an inducible promoter. Bioinformatics screening of the bacterial nucleotide database, shows that regions homologous to the Lpt TA locus are widely distributed in the Lactobacillus genus, particularly within the L. casei group, suggesting a relevant role of TA systems in plasmid maintenance of cheese microbiota.

  3. Can botulinum toxin type A injection technique influence the clinical outcome of patients with post-stroke upper limb spasticity? A randomized controlled trial comparing manual needle placement and ultrasound-guided injection techniques.

    PubMed

    Santamato, Andrea; Micello, Maria Francesca; Panza, Francesco; Fortunato, Francesca; Baricich, Alessio; Cisari, Carlo; Pilotto, Alberto; Logroscino, Giancarlo; Fiore, Pietro; Ranieri, Maurizio

    2014-12-15

    Botulinum toxin type A is a first-line treatment for post-stroke focal spasticity, and the accuracy in delivering the toxin to the target muscles may influence the treatment outcome. Our aim was to compare the reduction of spasticity and the related finger position at rest improvement in post-stroke patients treated with botulinum toxin type A in upper limb muscles using ultrasound guidance and manual needle placement. In a randomized clinical trial, two groups of 15 stroke patients were treated with botulinum toxin type A injections in the wrist and finger flexor muscles of the affected upper limb using ultrasound guidance or manual needle placement. The Modified Ashworth Scale and the finger position at rest were measured at baseline and one month after toxin injections. After one month of follow-up from toxin injections, the Modified Ashworth Scale and finger position at rest significantly improved in both treatment groups, although these clinical outcomes were significantly better in patients treated under ultrasound guidance than in patients injected using manual needle placement. Ultrasound guidance for botulinum toxin type A injections could improve clinical outcome measures better than manual needle placement in post-stroke patients with spasticity. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Preparation and characterization of solar exfoliated graphene

    SciTech Connect

    M, Sreejesh S, Nagaraja H.; K, Udaya Bhat

    2014-10-15

    Hummer's method was used for the chemical synthesis of graphite oxide from graphite flakes. Simultaneous exfoliation and reduction of graphite oxide to Graphene was achieved through focused solar light irradiation using a convex lens. The morphological characteristics were studied using SEM and TEM. Layered morphology of Graphene was observed through TEM. Raman spectra and FTIR were used for the structural characterization of Graphene. EDAX analysis showed the drop in oxygen content during exfoliation. The method offered a faster, easier and environmental friendly method to produce Graphene for potential applications.

  5. Method for exfoliation of hexagonal boron nitride

    NASA Technical Reports Server (NTRS)

    Lin, Yi (Inventor); Connell, John W. (Inventor)

    2012-01-01

    A new method is disclosed for the exfoliation of hexagonal boron nitride into mono- and few-layered nanosheets (or nanoplatelets, nanomesh, nanoribbons). The method does not necessarily require high temperature or vacuum, but uses commercially available h-BN powders (or those derived from these materials, bulk crystals) and only requires wet chemical processing. The method is facile, cost efficient, and scalable. The resultant exfoliated h-BN is dispersible in an organic solvent or water thus amenable for solution processing for unique microelectronic or composite applications.

  6. The device application of electrochemical exfoliated graphene

    NASA Astrophysics Data System (ADS)

    Tan, Chee Kiat; Beh, Khi Poay; Suhaimi, Faris Hidayat Ahmad; Ng, Yu Zhang; Yam, Fong Kwong; Lim, Hwee San; Jafri, Mohd. Zubir Mat

    2017-08-01

    In this work, graphene was exfoliated by introducing the constant Galvano Static Current (GSC) at the range of 300-600mA with the incremental of 100mA. The Graphene produced through this exfoliation technique was also investigated through optical characterization using Field Emission Scanning Electron Microscope (FESEM), Ultraviolet Visible (UV-vis) spectroscopy and Raman spectroscopy to comprehend the quality behavior. To further understand the graphene characteristic, we applied the produced graphene on top of fabricated interdigitated electrode (IDE) device to investigate the electrical reaction.

  7. Hypothalamic-pituitary-adrenal axis suppression related to topical glucocorticoid therapy in a child with psoriatic exfoliative erythroderma.

    PubMed

    Campbell, Lauren S; Chevalier, Michelle; Levy, Richard A; Rhodes, Arthur

    2012-01-01

    Exfoliative erythroderma is a rare presentation of psoriasis in children and adults. We report a 9-year-old girl with exfoliative erythroderma secondary to plaque-type psoriasis who developed hypothalamic-pituitary-adrenal axis suppression resulting from topical treatment with a medium-potency glucocorticoid. This case emphasizes the need for awareness of this potentially life-threatening complication of topical glucocorticoid use, particularly in patients who have significant compromise of barrier function secondary to widespread skin disease.

  8. Activity of botulinum toxin type A in cranial dura: implications for treatment of migraine and other headaches

    PubMed Central

    Filipović, Boris; Matak, Ivica; Helyes, Zsuzsanna

    2016-01-01

    Background and Purpose Although botulinum toxin type A (BoNT/A) is approved for chronic migraine treatment, its mechanism of action is still unknown. Dural neurogenic inflammation (DNI) commonly used to investigate migraine pathophysiology can be evoked by trigeminal pain. Here, we investigated the reactivity of cranial dura to trigeminal pain and the mechanism of BoNT/A action on DNI. Experimental Approach Because temporomandibular disorders are highly comorbid with migraine, we employed a rat model of inflammation induced by complete Freund's adjuvant, followed by treatment with BoNT/A injections or sumatriptan p.o. DNI was assessed by Evans blue‐plasma protein extravasation, cell histology and RIA for CGRP. BoNT/A enzymatic activity in dura was assessed by immunohistochemistry for cleaved synaptosomal‐associated protein 25 (SNAP‐25). Key Results BoNT/A and sumatriptan reduced the mechanical allodynia and DNI, evoked by complete Freund's adjuvant. BoNT/A prevented inflammatory cell infiltration and inhibited the increase of CGRP levels in dura. After peripheral application, BoNT/A‐cleaved SNAP‐25 colocalized with CGRP in intracranial dural nerve endings. Injection of the axonal transport blocker colchicine into the trigeminal ganglion prevented the formation of cleaved SNAP‐25 in dura. Conclusions and Implications Pericranially injected BoNT/A was taken up by local sensory nerve endings, axonally transported to the trigeminal ganglion and transcytosed to dural afferents. Colocalization of cleaved SNAP‐25 and the migraine mediator CGRP in dura suggests that BoNT/A may prevent DNI by suppressing transmission by CGRP. This might explain the effects of BoNT/A in temporomandibular joint inflammation and in migraine and some other headaches. PMID:26493010

  9. Botulinum Toxin Type A for Cephalic Cutaneous Allodynia in Chronic Migraine: A Randomized, Double-Blinded, Placebo-Controlled Trial

    PubMed Central

    Hollanda, Luciano; Monteiro, Larissa; Melo, Ailton

    2014-01-01

    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. PMID:25568735

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

    PubMed

    Unlu, Ece; Cevikol, Alev; Bal, Burcu; Gonen, Emel; Celik, Ozlem; Kose, Gulşen

    2010-06-01

    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. 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. 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. We found that the Ashworth and Tardieu scores decreased significantly at the three-month evaluation (p<0.05) but not at the six-month evaluation (p>0.05). 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. We believe that a single multilevel BTX-A injection reduces spasticity and improves motor function in children with cerebral palsy.

  11. [Clinical effect of ultrasound combined with anatomy-measure guidance botulinum toxin type A injection in poststroke patients with spasticity].

    PubMed

    JIANG, Li; WEI, Xiao-mei; DOU, Zu-lin; LI, Xin; WANG, Qiao-yuan; LAN, Yue; HU, Xi-quan; ZHENG, Hai-qing

    2012-04-17

    To evaluate the clinical efficacies of botulinum toxin type A (BTX-A) injection under ultrasonic guidance and body surface positioning in poststroke patients with lower extremities spasticity. From January 2009 to January 2011, a total of 18 patients with stroke-related spasticity in lower extremities were recruited at Third Affiliated Hospital of Sun Yat-sen University. Under the guide of color Doppler ultrasound and body surface positioning, BTX-A was injected into multi-points of muscles. The outcome after BTX-A injection was assessed by modified Ashworth scale (MAS), passive range of movement (PROM), 10-meter walking test (10 MWT) and Berg balance scale (BBS). Assessments were performed at baseline, Day 3, Weeks 1, 2, 4 and 12 post-injection respectively. Compared the scores of MAS (MAS(pre-treatment) 2.6 ± 0.5, MAS(post-treatment) 1.9 ± 0.2 - 1.1 ± 0.3 score), PROM (PROM(pre-treatment) 7.2 ± 2.4°, PROM(post-treatment) 12.3 ± 2.0 - 18.6 ± 2.2°) between baseline and follow-up at Weeks 1, 2, 4 and 12 post-treatment, there were significant statistical differences (P < 0.05).10 MW (10 MWT(pre-treatment) 55.1 ± 5.2 s, 10 MWT(post-treatment) 48.6 ± 4.2 - 42.9 ± 3.8 s) and BBS (BBS(pre-treatment) 34.7 ± 5.1, BBS(post-treatment) 39.9 ± 4.9 - 45.8 ± 2.1 score) improved greatly at Weeks 2, 4 and 12 post-treatment. Ultrasonic guidance and body surface positioning is an accurate positioning modality of using BTX-A for treating the spasticity of lower extremities.

  12. Multilevel Botulinum Toxin Type A as a Treatment for Spasticity in Children with Cerebral Palsy: A Retrospective Study

    PubMed Central

    Unlu, Ece; Cevikol, Alev; Bal, Burcu; Gonen, Emel; Celik, Ozlem; Kose, Gulşen

    2010-01-01

    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 (p<0.05) but not at the six-month evaluation (p>0.05). 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. PMID:20613938

  13. Botulinum toxin type A injections can be an effective treatment for pain in children with hip spasms and cerebral palsy.

    PubMed

    Lundy, Claire T; Doherty, Gary M; Fairhurst, Charlie B

    2009-09-01

    Botulinum toxin type A (BoNT-A) injections were used in the treatment of lower-limb spasticity in children with cerebral palsy (CP). Anecdotal evidence suggests a reduction in pain after this treatment in children who had pain localized to a displaced hip joint. We report on our current clinical practice. Twenty-six children with non-ambulant quadriplegic CP (Gross Motor Function Classification System level V) were assessed as having significant spasticity and pain at the hip level. Twelve were males and 14 females, with an age range of 2 to 19 years (mean age 11y 6mo, SD 4y 9mo). Ten had functional difficulties secondary to predominant spasticity and 16 had a mix of a high-background peripheral tone with superimposed dystonia. Of the 26 children assessed, 10 had at least one hip which was dislocated and three had at least one hip which was subluxed. As part of their spasticity management programme they received targeted BoNT-A injections to the adductor magnus, medial hamstrings and iliopsoas muscle groups. The Paediatric Pain Profile was used as the primary outcome measure. All had highly significant improvement in their recorded pain profile scores measured at 3 months after treatment (p<0.001). There was equal efficacy in response to treatment in the children with subluxed or dislocated hips. In addition, families commented on improved quality of life for the children across several areas, including sleep, postural management, and activities of daily living. This report demonstrates that targeted BoNT-A injections reduced pain in children with significant spasticity and pain at the hip level. They may also improve quality of life of non-ambulant children with CP and a hip problem.

  14. Cortical somatosensory evoked potentials and spasticity assessment after botulinum toxin type A injection in children with cerebral palsy.

    PubMed

    Boćkowski, L; Okurowska-Zawada, B; Sobaniec, W; Kułak, W; Sendrowski, K

    2007-01-01

    The mechanism of Botulinum Toxin Type A (BTX-A) action at the neuromuscular junction is well known. But from the introduction of BTX-A, some authors have suggested a central action of BTX-A and possible side effects far from the site of injection. Some studies demonstrate an improvement of cortical SEPs associated with reduction of spasticity after BTX-A injection. The aim of the present study was to determine the effect of BTX-A treatment on cortical somatosensory potentials (SEP). A group of twenty nine children ranging from 2 to 17 years old with cerebral palsy were studied. Each patients spasticity level was evaluated before, 2 weeks and 6 weeks after BTX-A injection by the Modified Ashworth Scale and modified Gait Physician's Rating Scale. The SEPs from lower and upper extremities were performed before and between 2 and 6 weeks (19.34 +/- 8.82 days) after BTX-A administration. The mean spasitity level was significantly lower 2 and 6 weeks after BTX-A injection. The gait analysis by modified Physician's Rating Scale (PRS) showed significant improvement two weeks and six weeks after BTX-A injection. SEPs results were abnormal before BTX-A injection in 25 children with cerebral palsy. However we didn't find any significant changes of SEPs latencies after BTX-A injection. The results of SEP after BTX-A administration in children with cerebral palsy do not confirm the central action of BTX-A on somatosensory pathways. We did not find any significant changes of SEP latencies associated with clinical reduction of spasticity. It seems that SEP results could support the opinion, that BTX-A does not have any direct central effect on sensory pathways. Remote side effects may be explained by an indirect mechanism due to modification of the central loops of reflexes or to hematogenous spread of BTX-A.

  15. The Ess/Type VII secretion system of Staphylococcus aureus secretes a nuclease toxin that targets competitor bacteria

    PubMed Central

    Cao, Zhenping; Casabona, M. Guillermina; Kneuper, Holger; Chalmers, James D.; Palmer, Tracy

    2017-01-01

    Summary The type VII protein secretion system (T7SS) plays a critical role in the virulence of human pathogens including Mycobacterium tuberculosis and Staphylococcus aureus. Here we report that the S. aureus T7SS secretes a large nuclease toxin, EsaD. The toxic activity of EsaD is neutralised during its biosynthesis through complex formation with an antitoxin, EsaG, which binds to its C-terminal nuclease domain. The secretion of EsaD is dependent upon a further accessory protein, EsaE, that does not interact with the nuclease domain, but instead binds to the EsaD N-terminal region. EsaE has a dual cytoplasmic/membrane localization and membrane-bound EsaE interacts with the T7SS secretion ATPase, EssC, implicating EsaE in targeting the EsaDG complex to the secretion apparatus. EsaD and EsaE are co-secreted whereas EsaG is found only in the cytoplasm and may be stripped off during the secretion process. Strain variants of S. aureus that lack esaD encode at least two copies of EsaG-like proteins most likely to protect themselves from the toxic activity of EsaD secreted by esaD+ strains. In support of this, a strain overproducing EsaD elicits significant growth inhibition against a sensitive strain. We conclude that T7SSs may play unexpected and key roles in bacterial competitiveness. PMID:27723728

  16. The Effect of Botulinum Toxin Type A on Expression Profiling of Long Noncoding RNAs in Human Dermal Fibroblasts

    PubMed Central

    Miao, Ying-ying; Liu, Juan; Zhu, Jie; Tao, Yan-ling; Zhang, Jia-an

    2017-01-01

    Objective. This study was aimed at analyzing the expressions of long noncoding RNAs (lncRNAs) in Botulinum Toxin Type A (BoNTA) treated human dermal fibroblasts (HDFs) in vitro. Methods. We used RNA sequencing to characterize the lncRNAs and mRNAs transcriptome in the control and BoNTA treated group, in conjunction with application of GO (gene ontology) analysis and KEGG (kyoto encyclopedia of genes and genomes) analysis to delineate the alterations in gene expression. We also obtained quantitative real time polymerase chain reaction (qRT-PCR) to confirm some differentially expressed genes. Results. Numerous differentially expressed genes were observed by microarrays between the two groups. qRT-PCR confirmed the changes of six lncRNAs (RP11-517C16.2-001, FR271872, LOC283352, RP11-401E9.3, FGFR3P, and XXbac-BPG16N22.5) and nine mRNAs (NOS2, C13orf15, FOS, FCN2, SPINT1, PLAC8, BIRC5, NOS2, and COL19A1). Farther studies indicated that the downregulating effect of BoNTA on the expression of FGFR3P was time-related and the dosage of BoNTA at a range from 2.5 U/106 cells to 7.5 U/106 cells increased the expression of FGFR3P and COL19A1 in HDFs as well. Conclusion. The expression profiling of lncRNAs was visibly changed in BoNTA treated HDFs. Further studies should focus on several lncRNAs to investigate their functions in BoNTA treated HDFs and the underlying mechanisms. PMID:28265570

  17. International consensus recommendations on the aesthetic usage of botulinum toxin type A (Speywood Unit)--Part I: Upper facial wrinkles.

    PubMed

    Ascher, B; Talarico, S; Cassuto, D; Escobar, S; Hexsel, D; Jaén, P; Monheit, G D; Rzany, B; Viel, M

    2010-11-01

    Azzalure (Galderma SA) is a newly approved European botulinum neurotoxin type A (BoNT-A). It is derived from Dysport (Ipsen Pharma), which has a long history of usages in various applications. Azzalure and Dysport are collectively referred to as BoNT-A (Speywood Unit) and are different from other BoNT-A preparations. To provide consensus recommendations on the treatment of upper face wrinkles with BoNT-A (Speywood Unit). The members of the International Board on Botulinum toxin Azzalure (IBBA) convened to develop consensus on the treatment of upper facial wrinkles based on their own extensive experience. The consensus recommendations address the general issues regarding treatment and provide specific guidelines on the anatomy, injection points, dose, injection technique and safety precautions concerning each common upper face indication. The recommended final concentration of BoNT-A (Speywood Unit) is 200 s.U/mL (10 s.U/0.05 mL) after reconstitution. For glabellar lines, the members recommend a total of five injection points with 10 s.U/point. For forehead wrinkles, the members recommend four to six injections into the frontalis with 5-10 s.U/point. For crow's feet, the members recommend three injections per side with 5-10 s.U/point at the lateral part of the orbicularis oculi. For lateral eyebrow lift, the members recommend one point at each eyebrow tail and an additional one in each side of the frontalis with 5-10 s.U/point. This guideline provides a framework for physicians who wish to perform safe and efficacious injection of BoNT-A (Speywood Unit). © 2010 The Authors. Journal of the European Academy of Dermatology and Venereology © 2010 European Academy of Dermatology and Venereology.

  18. Relevance of neutrophils in the murine model of haemolytic uraemic syndrome: mechanisms involved in Shiga toxin type 2-induced neutrophilia.

    PubMed

    Fernandez, G C; Lopez, M F; Gomez, S A; Ramos, M V; Bentancor, L V; Fernandez-Brando, R J; Landoni, V I; Dran, G I; Meiss, R; Isturiz, M A; Palermo, M S

    2006-10-01

    It has been demonstrated that infections due to Shiga toxins (Stx) producing Escherichia coli are the main cause of the haemolytic uraemic syndrome (HUS). However, the contribution of the inflammatory response in the pathogenesis of the disease has also been well established. Neutrophils (PMN) represent a central component of inflammation during infections, and patients with high peripheral PMN counts at presentation have a poor prognosis. The mouse model of HUS, by intravenous injection of pure Stx type 2 (Stx2), reproduces human neutrophilia and allows the study of early events in the course of Stx2-induced pathophysiological mechanisms. The aim of this study was to address the contribution of PMN on Stx2 toxicity in a murine model of HUS, by evaluating the survival and renal damage in mice in which the granulocytic population was depleted. We found that the absence of PMN reduced Stx2-induced lethal effects and renal damage. We also investigated the mechanisms underlying Stx2-induced neutrophilia, studying the influence of Stx2 on myelopoyesis, on the emergence of cells from the bone marrow and on the in vivo migration into tissues. Stx2 administration led to an accelerated release of bone marrow cells, which egress at an earlier stage of maturation, together with an increase in the proliferation of myeloid progenitors. Moreover, Stx2-treated mice exhibited a lower migratory capacity to a local inflammatory site. In conclusion, PMN are essential in the pathogenesis of HUS and neutrophilia is not merely an epiphenomenon, but contributes to Stx2-damaging mechanism by potentiating Stx2 toxicity.

  19. The Effect of Botulinum Toxin Type A on Expression Profiling of Long Noncoding RNAs in Human Dermal Fibroblasts.

    PubMed

    Miao, Ying-Ying; Liu, Juan; Zhu, Jie; Tao, Yan-Ling; Zhang, Jia-An; Luo, Dan; Zhou, Bing-Rong

    2017-01-01

    Objective. This study was aimed at analyzing the expressions of long noncoding RNAs (lncRNAs) in Botulinum Toxin Type A (BoNTA) treated human dermal fibroblasts (HDFs) in vitro. Methods. We used RNA sequencing to characterize the lncRNAs and mRNAs transcriptome in the control and BoNTA treated group, in conjunction with application of GO (gene ontology) analysis and KEGG (kyoto encyclopedia of genes and genomes) analysis to delineate the alterations in gene expression. We also obtained quantitative real time polymerase chain reaction (qRT-PCR) to confirm some differentially expressed genes. Results. Numerous differentially expressed genes were observed by microarrays between the two groups. qRT-PCR confirmed the changes of six lncRNAs (RP11-517C16.2-001, FR271872, LOC283352, RP11-401E9.3, FGFR3P, and XXbac-BPG16N22.5) and nine mRNAs (NOS2, C13orf15, FOS, FCN2, SPINT1, PLAC8, BIRC5, NOS2, and COL19A1). Farther studies indicated that the downregulating effect of BoNTA on the expression of FGFR3P was time-related and the dosage of BoNTA at a range from 2.5 U/10(6) cells to 7.5 U/10(6) cells increased the expression of FGFR3P and COL19A1 in HDFs as well. Conclusion. The expression profiling of lncRNAs was visibly changed in BoNTA treated HDFs. Further studies should focus on several lncRNAs to investigate their functions in BoNTA treated HDFs and the underlying mechanisms.

  20. An Evaluation of Use of Botulinum Toxin Type A in the Management of Dynamic Forehead Wrinkles - A Clinical Study

    PubMed Central

    Susmita, Avvaru; Kolli, Naga Neelima Devi; Meka, Sridhar; Chakravarthi, Srinivas Pandi; Lingamaneni, Krishna Prasad; Shaik, Latheef Saheb

    2016-01-01

    Introduction The pursuit of youth and beauty has undergone a resurgence of interest which is evidenced by increasing cosmetic procedures. Botulinum Toxin Type A (Botox) is one among the many procedures invented for facial rejuvenation which denervates certain muscles of facial expression responsible for facial wrinkles. It has been applied in the forehead, glabella, lateral canthal area and neck. In maxillofacial area hyperactive forehead wrinkles show sagging. Aim This study was aimed to clinically evaluate the efficacy of Botox injection in the elimination of hyperdynamic forehead wrinkles and the objectives were to compare pre-operative and post-operative improvement in the number of wrinkles, photographic grading and patient satisfaction responses after 1st week, 4th week and 16th week. Materials and Methods A total of 10 patients were randomly included in the present study who were cooperative, motivated and aesthetically conscious with moderate to severe forehead wrinkles. Assessment was performed clinically, photographically (using standardized photographs) and patient satisfaction responses were recorded at 1st week, 4th week and 16th week. Results The study showed a significant difference in the elimination of wrinkles at rest and in action when assessed at 1st week and 4th week and it was consistent at 16th week. The patient showed positive satisfaction response without ptosis of the upper eyelid. Conclusion Treatment with Botox is simple, safe and an effective modality for reduction of forehead wrinkles. It offers an alternative management in a cost-effective way when compared to surgical procedures. PMID:27891474

  1. Botulinum toxin type A (Botox) for the neuromuscular correction of excessive gingival display on smiling (gummy smile).

    PubMed

    Polo, Mario

    2008-02-01

    Previously, botulinum toxin type A (BTX-A) (Botox; Allergan, Irvine, Calif) was shown to be effective in reducing excessive gingival display in 5 patients with gummy smiles. This study was conducted to determine whether the doses and the primary injection sites used in the pilot study for the correction of gummy smiles provide consistent, statistically significant, and esthetically pleasing results. Thirty patients received BTX-A injections to reduce excessive gingival display. Gingival display was defined as the difference between the lower margin of the upper lip and the superior margin of the right incisor. Patients were followed at 2, 4, 8, 12, 16, 20, and 24 weeks postinjection, with changes documented by photographs and videos. At week 2, the patients rated the effects of BTX-A. A group of specialty clinicians also evaluated the effects of BTX-A. Preinjection gingival display averaged 5.2 +/- 1.4 mm in the 30 patients. At 2 weeks postinjection, mean gingival display had declined to 0.09 mm (+/- 1.06 mm) in 30 patients (t = 26.01, P <.00001). The average lip-drop at 2 weeks was 5.1 mm for 30 patients. Gingival display gradually increased from 2 weeks postinjection through 24 weeks, but, at 24 weeks, average gingival display had not returned to baseline values. Based on predictions from a third-order polynomial equation, the baseline average of 5.2 mm would not be reached until 30 to 32 weeks postinjection. Patients and specialty evaluators rated the effects of BTX-A as highly favorable. BTX-A injections for the neuromuscular correction of gummy smiles caused by hyperfunctional upper lip elevator muscles was effective and statistically superior to baseline smiles, although the effect is transitory.

  2. Botulinum Toxin Type A Inhibits α-Smooth Muscle Actin and Myosin II Expression in Fibroblasts Derived From Scar Contracture.

    PubMed

    Chen, Minliang; Yan, Tongtong; Ma, Kui; Lai, Linying; Liu, Chang; Liang, Liming; Fu, Xiaobing

    2016-09-01

    Scar contracture (SC) is one of the most common complications resulting from major burn injuries. Numerous treatments are currently available but they do not always yield excellent therapeutic results. Recent reports suggest that botulinum toxin type A (BTXA) is effective at reducing SC clinically, but the molecular mechanism for this action is unknown. α-Smooth muscle actin (α-SMA) and myosin II are the main components of stress fibers, which are the contractile structures of fibroblasts. The effects of BTXA on α-SMA and myosin II in SC are still unknown. This study aimed to explore the effect of BTXA on α-SMA and myosin II expression in fibroblasts derived from SC and to elucidate its actual mechanism further. Fibroblasts were isolated from tissue specimens of SC. Fibroblasts were cultured in Dulbecco modified Eagle medium with different concentrations of BTXA and their proliferation was analyzed through the tetrazolium-based colorimetric method at 1, 4, and 7 days. Proteins of α-SMA and myosin II were checked using Western blot in fibroblasts treated with different concentrations of BTXA at 1, 4, and 7 days. Fibroblasts without BTXA treatment had a higher proliferation than that in other groups, which indicated that the proliferation of fibroblasts was significantly inhibited by BTXA (P < 0.05). Proteins of α-SMA and myosin II between fibroblasts with BTXA and fibroblasts without BTXA are statistically significant (P < 0.05). These results suggest that BTXA effectively inhibited the growth of fibroblasts derived from SC and reduced the expression of α-SMA and myosin II, which provided theoretical support for the application of BTXA to control SC.

  3. Efficacy and safety of botulinum toxin type A for treatment of Frey’s syndrome: evidence from 22 published articles

    PubMed Central

    Xie, Shang; Wang, Kan; Xu, Tao; Guo, Xue-Sheng; Shan, Xiao-Feng; Cai, Zhi-Gang

    2015-01-01

    Frey’s syndrome (FS) is an unavoidable sequela following the surgery of the parotid gland. Although several treatment methods are available, their efficacy is short term or accompanied by unacceptable complications. In the past two decades, botulinum toxin type A (BTXA) has been widely used to treat FS. Although several systematic reviews have been reported recently, they were conflicting and with obvious deficiencies. Thus, we performed an objectively systematic review to determine whether BTXA is an effective and safe treatment for FS. A literature retrieval covering PubMed, Web of Science, Ovid, Embase and Cochrane library was performed on 16 January, 2015. Proportion meta-analysis and corresponding 95% confidence interval (CI) were performed to evaluate the efficacy and safety of BXTA in treatment of FS. A total of 499 records were retrieved and 22 articles with 23 studies were included after scrutiny by two independent authors. Statistical analyses regarding the effective rate, incidence of complications were used to estimate the efficacy and safety of BTXA. Our results suggested that the effective rate of BTXA for treatment of FS is 98.5% (95% CI = 0.971–0.994) and the incidence of complication is 3.6% (95% CI = 0.017–0.061). In conclusion, our study supports that BTXA produces meaningful benefits on the treatment of patients with FS. However, owing to lack of strong evidence, future studies with well-designed inclusion criteria and multicenter randomized controlled trials are needed to give more credible evidence, if possible. PMID:26310612

  4. A PCR-RFLP assay to detect and type cytolethal distending toxin (cdt) genes in Campylobacter hyointestinalis

    PubMed Central

    HATANAKA, Noritoshi; KAMEI, Kazumasa; SOMROOP, Srinuan; AWASTHI, Sharda Prasad; ASAKURA, Masahiro; MISAWA, Naoaki; HINENOYA, Atsushi; YAMASAKI, Shinji

    2016-01-01

    Campylobacter hyointestinalis is considered as an emerging zoonotic pathogen. We have recently identified two types of cytolethal distending toxin (cdt) gene in C. hyointestinalis and designated them as Chcdt-I and Chcdt-II. In this study, we developed a PCR-restriction fragment length polymorphism (RFLP) assay that can differentiate Chcdt-I from Chcdt-II. When the PCR-RFLP assay was applied to 17 other Campylobacter strains and 25 non-Campylobacter strains, PCR products were not obtained irrespective of their cdt gene-possession, indicating that the specificity of the PCR-RFLP assay was 100%. In contrast, when the PCR-RFLP assay was applied to 35 C. hyointestinalis strains including 23 analyzed in the previous study and 12 newly isolated from pigs and bovines, all of them showed the presence of cdt genes. Furthermore, a restriction digest by EcoT14-I revealed that 29 strains contained both Chcdt-I and Chcdt-II and 6 strains contained only Chcdt-II, showing 100% sensitivity. Unexpectedly, however, PCR products obtained from 7 C. hyointestinalis strains were not completely digested by EcoT14-I. Nucleotide sequence analysis revealed that the undigested PCR product was homologous to cdtB but not to Chcdt-IB or Chcdt-IIB, indicating the presence of another cdt gene-variant. Then, we further digested the PCR products with DdeI in addition to EcoT14-I, showing that all three cdt genes, including a possible new Chcdt variant, could be clearly differentiated. Thus, the PCR-RFLP assay developed in this study is a valuable tool for evaluating the Chcdt gene-profile of bacteria. PMID:27916784

  5. Diversity of Fusarium head blight populations and trichothecene toxin types reveals regional differences in pathogen composition and temporal dynamics.

    PubMed

    Kelly, Amy C; Clear, Randall M; O'Donnell, Kerry; McCormick, Susan; Turkington, T Kelly; Tekauz, Andy; Gilbert, Jeannie; Kistler, H Corby; Busman, Mark; Ward, Todd J

    2015-09-01

    Analyses of genetic diversity, trichothecene genotype composition, and population structure were conducted using 4086 Fusarium graminearum isolates collected from wheat in eight Canadian provinces over a three year period between 2005 and 2007. The results revealed substantial regional differences in Fusarium head blight pathogen composition and temporal population dynamics. The 3ADON trichothecene type consistently predominated in Maritime provinces (91%) over the sampled years, and increased significantly (P<0.05) between 2005 and 2007 in western Canada, accounting for 66% of the isolates in Manitoba by the end of the sampling period. In contrast, 3ADON frequency was lower (22%, P<0.001) in the eastern Canadian provinces of Ontario and Québec and did not change significantly between 2005 and 2007, resulting in two distinct longitudinal clines in 3ADON frequency across Canada. Overall, genetic structure was correlated with toxin type, as the endemic population (NA1) was dominated by 15ADON isolates (86%), whereas a second population (NA2) consisted largely of 3ADON isolates (88%). However, the percentage of isolates with trichothecene genotypes that were not predictive of their genetic population assignment (recombinant genotypes) increased from 10% in 2005 to 17% in 2007, indicating that trichothecene type became an increasingly unreliable marker of population identity over time. In addition, there were substantial regional differences in the composition of recombinant genotypes. In western and maritime provinces, NA2 isolates with 15ADON genotypes were significantly more common than NA1 isolates with 3ADON genotypes (P<0.001), and the reverse was true in the eastern provinces of Québec and Ontario. Temporal trends in recombinant genotype composition also varied regionally, as the percentage of 15ADON isolates with NA2 genetic backgrounds increased approximately three fold in western and Maritime provinces, while the opposite trends were observed in Québec and

  6. A study on the cause of death produced by angusticeps-type toxin F7 isolated from eastern green mamba venom.

    PubMed

    Lee, C Y; Lee, S Y; Chen, Y M

    1986-01-01

    The cause of death due to toxin F7, an angusticeps-type toxin, isolated from the venom of Dendroaspis angusticeps was studied in anesthetized mice. The carotid arterial blood pressure, the ECG and the respiratory movements were recorded. Within a few minutes after i.v. injection of F7 (1 mg/kg), both the rate and amplitude of the respiratory movements decreased and respiratory arrest took place within 15 min in most cases. Before respiratory arrest, marked bradycardia with various types of arrhythmia and oscillation of blood pressure were observed. Artificial ventilation could abolish these cardiovascular changes and maintain the blood pressure for a long period. Toxin F7 caused a transient and slight increase of arterial blood pressure which could be prevented by hexamethonium. Intracisternal application of F7 (1 mg/kg) caused a long-lasting hypertension and bradycardia and the respiratory arrest time was significantly longer than after i.v. injection. A large dose (50 mg/kg i.p.) of atropine, but not smaller doses (5-10 mg/kg), protected mice against respiratory failure induced by F7. In rats, the phrenic nerve discharge was prolonged during respiratory depression. Since F7 has a potent anticholinesterase activity, it is concluded that the respiratory failure induced by F7 is peripheral in origin, chiefly, if not entirely, due to its anticholinesterase activity.

  7. Variability of antibiotic susceptibility and toxin production of Staphylococcus aureus strains isolated from skin, soft tissue, and bone related infections

    PubMed Central

    2013-01-01

    Background Staphylococcus aureus is an opportunistic commensal bacterium that mostly colonizes the skin and soft tissues. The pathogenicity of S. aureus is due to both its ability to resist antibiotics, and the production of toxins. Here, we characterize a group of genes responsible for toxin production and antibiotic resistance of S. aureus strains isolated from skin, soft tissue, and bone related infections. Results A total of 136 S. aureus strains were collected from five different types of infection: furuncles, pyomyositis, abscesses, Buruli ulcers, and osteomyelitis, from hospital admissions and out-patients in Benin. All strains were resistant to benzyl penicillin, while 25% were resistant to methicillin, and all showed sensitivity to vancomycin. Panton-Valentine leukocidin (PVL) was the most commonly produced virulence factor (70%), followed by staphylococcal enterotoxin B (44%). Exfoliative toxin B was produced by 1.3% of the strains, and was only found in isolates from Buruli ulcers. The tsst-1, sec, and seh genes were rarely detected (≤1%). Conclusions This study provides new insight into the prevalence of toxin and antibiotic resistance genes in S. aureus strains responsible for skin, soft tissue, and bone infections. Our results showed that PVL was strongly associated with pyomyositis and osteomyelitis, and that there is a high prevalence of PVL-MRSA skin infections in Benin. PMID:23924370

  8. Immunization with recombinant bivalent chimera r-Cpae confers protection against alpha toxin and enterotoxin of Clostridium perfringens type A in murine model.

    PubMed

    Shreya, Das; Uppalapati, Siva R; Kingston, Joseph J; Sripathy, Murali H; Batra, Harsh V

    2015-05-01

    Clostridium perfringens type A, an anaerobic pathogen is the most potent cause of soft tissue infections like gas gangrene and enteric diseases like food poisoning and enteritis. The disease manifestations are mediated via two important exotoxins, viz. myonecrotic alpha toxin (αC) and enterotoxin (CPE). In the present study, we synthesized a bivalent chimeric protein r-Cpae comprising C-terminal binding regions of αC and CPE using structural vaccinology rationale and assessed its protective efficacy against both alpha toxin (αC) and enterotoxin (CPE) respectively, in murine model. Active immunization of mice with r-Cpae generated high circulating serum IgG (systemic), significantly increased intestinal mucosal s-IgA antibody titres and resulted in substantial protection to the immunized animals (100% and 75% survival) with reduced tissue morbidity when administered with 5×LD(100) doses of αC (intramuscular) and CPE (intra-gastric gavage) respectively. Mouse RBCs and Caco-2 cells incubated with a mixture of anti-r-Cpae antibodies and αC and CPE respectively, illustrated significantly higher protection against the respective toxins. Passive immunization of mice with a similar mixture resulted in 91-100% survival at the end of the 15 days observation period while mice immunized with a concoction of sham sera and respective toxins died within 2-3 days. This work demonstrates the efficacy of the rationally designed r-Cpae chimeric protein as a potential sub unit vaccine candidate against αC and CPE of C. perfringens type A toxemia. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Botulinum toxin type A targets RhoB to inhibit lysophosphatidic acid-stimulated actin reorganization and acetylcholine release in nerve growth factor-treated PC12 cells.

    PubMed

    Ishida, Hiroshi; Zhang, Xieping; Erickson, Kelly; Ray, Prabhati

    2004-09-01

    Botulinum toxin type A (BoNT/A) produced by Clostridium botulinum inhibits Ca2+-dependent acetylcholine (ACh) release (neuroexocytosis) at peripheral neuromuscular junctions, sometimes causing neuromuscular paralysis. This inhibitory effect is attributed to its metalloprotease activity to cleave the 25-kDa synaptosomal-associated protein, which is essential for the exocytotic machinery. However, deletion of this protein does not result in a complete block of neuroexocytosis, suggesting that botulinum-mediated inhibition may occur via another mechanism. Rho GTPases, a class of small GTP-binding proteins (G proteins), control actin cytoskeletal organization, thereby regulating a variety of cellular functions in various cells, including neuronal cells. We have shown that the G protein activator lysophosphatidic acid (LPA) triggered actin reorganization followed by Ca2+-dependent ACh release in nerve growth factor-treated PC12 cells and that BoNT/A blocked both events through degradation of RhoB by the proteasome. Overexpression of wild-type RhoB caused actin reorganization and enhanced the release of ACh by LPA to overcome toxin's inhibitory effect on actin reorganization/exocytosis stimulated by LPA, whereas overexpression of a dominant negative RhoB inhibited ACh release, regardless of LPA and/or toxin treatment. Finally, a knockdown of the RhoB gene via sequence-specific, post-transcriptional gene silencing reduced RhoB expression in PC12 cells, resulting in total inhibition of both actin reorganization and ACh release induced by LPA. We conclude that the RhoB signaling pathway regulates ACh release via actin cytoskeletal reorganization and that botulinum toxin inhibits neuroexocytosis by targeting RhoB pathway.

  10. Exfoliation of natural van der Waals heterostructures to a single unit cell thickness

    NASA Astrophysics Data System (ADS)

    Velický, Matěj; Toth, Peter S.; Rakowski, Alexander M.; Rooney, Aidan P.; Kozikov, Aleksey; Woods, Colin R.; Mishchenko, Artem; Fumagalli, Laura; Yin, Jun; Zólyomi, Viktor; Georgiou, Thanasis; Haigh, Sarah J.; Novoselov, Kostya S.; Dryfe, Robert A. W.

    2017-02-01

    Weak interlayer interactions in van der Waals crystals facilitate their mechanical exfoliation to monolayer and few-layer two-dimensional materials, which often exhibit striking physical phenomena absent in their bulk form. Here we utilize mechanical exfoliation to produce a two-dimensional form of a mineral franckeite and show that the phase segregation of chemical species into discrete layers at the sub-nanometre scale facilitates franckeite's layered structure and basal cleavage down to a single unit cell thickness. This behaviour is likely to be common in a wider family of complex minerals and could be exploited for a single-step synthesis of van der Waals heterostructures, as an alternative to artificial stacking of individual two-dimensional crystals. We demonstrate p-type electrical conductivity and remarkable electrochemical properties of the exfoliated crystals, showing promise for a range of applications, and use the density functional theory calculations of franckeite's electronic band structure to rationalize the experimental results.

  11. Exfoliation of natural van der Waals heterostructures to a single unit cell thickness

    PubMed Central

    Velický, Matěj; Toth, Peter S.; Rakowski, Alexander M.; Rooney, Aidan P.; Kozikov, Aleksey; Woods, Colin R.; Mishchenko, Artem; Fumagalli, Laura; Yin, Jun; Zólyomi, Viktor; Georgiou, Thanasis; Haigh, Sarah J.; Novoselov, Kostya S.; Dryfe, Robert A. W.

    2017-01-01

    Weak interlayer interactions in van der Waals crystals facilitate their mechanical exfoliation to monolayer and few-layer two-dimensional materials, which often exhibit striking physical phenomena absent in their bulk form. Here we utilize mechanical exfoliation to produce a two-dimensional form of a mineral franckeite and show that the phase segregation of chemical species into discrete layers at the sub-nanometre scale facilitates franckeite's layered structure and basal cleavage down to a single unit cell thickness. This behaviour is likely to be common in a wider family of complex minerals and could be exploited for a single-step synthesis of van der Waals heterostructures, as an alternative to artificial stacking of individual two-dimensional crystals. We demonstrate p-type electrical conductivity and remarkable electrochemical properties of the exfoliated crystals, showing promise for a range of applications, and use the density functional theory calculations of franckeite's electronic band structure to rationalize the experimental results. PMID:28194026

  12. Exfoliation of natural van der Waals heterostructures to a single unit cell thickness.

    PubMed

    Velický, Matěj; Toth, Peter S; Rakowski, Alexander M; Rooney, Aidan P; Kozikov, Aleksey; Woods, Colin R; Mishchenko, Artem; Fumagalli, Laura; Yin, Jun; Zólyomi, Viktor; Georgiou, Thanasis; Haigh, Sarah J; Novoselov, Kostya S; Dryfe, Robert A W

    2017-02-13

    Weak interlayer interactions in van der Waals crystals facilitate their mechanical exfoliation to monolayer and few-layer two-dimensional materials, which often exhibit striking physical phenomena absent in their bulk form. Here we utilize mechanical exfoliation to produce a two-dimensional form of a mineral franckeite and show that the phase segregation of chemical species into discrete layers at the sub-nanometre scale facilitates franckeite's layered structure and basal cleavage down to a single unit cell thickness. This behaviour is likely to be common in a wider family of complex minerals and could be exploited for a single-step synthesis of van der Waals heterostructures, as an alternative to artificial stacking of individual two-dimensional crystals. We demonstrate p-type electrical conductivity and remarkable electrochemical properties of the exfoliated crystals, showing promise for a range of applications, and use the density functional theory calculations of franckeite's electronic band structure to rationalize the experimental results.

  13. Percolation scaling in composites of exfoliated MoS2 filled with nanotubes and graphene.

    PubMed

    Cunningham, Graeme; Lotya, Mustafa; McEvoy, Niall; Duesberg, Georg S; van der Schoot, Paul; Coleman, Jonathan N

    2012-10-21

    Applications of films of exfoliated layered compounds in many areas will be limited by their relatively low electrical conductivity. To address this, we have prepared and characterised composites of a nano-conductor (nanotubes or graphene) embedded in a matrix of exfoliated MoS(2) nanosheets. Solvent exfoliation of MoS(2) nanosheets, followed by blending with dispersions of graphene or nanotubes allowed the formation of such composite films by vacuum filtration. This gave spatially uniform mixtures with fully tuneable nano-conductor content. By addition of the nano-conducting phase, it was possible to vary the electrical conductivity of the composite over nine orders of magnitude. For both filler types the conductivity followed percolation scaling laws both above and below the percolation threshold. In the case of SWNT-filled composites, conductivities as high as ~40 S m(-1) were achieved at volume fractions as low as ~4%.

  14. Oxcarbazepine-induced drug rash with eosinophilia and systemic symptoms syndrome presenting as exfoliative dermatitis.

    PubMed

    Saha, Mahimanjan; Gorai, Surajit; Madhab, Vaswatee

    2016-01-01

    Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome is a type of severe adverse cutaneous drug reaction characterized by fever, skin eruption, hematological abnormalities, and internal organ involvement. Although anticonvulsant drugs are mainly implicated in DRESS, newer anticonvulsants such as oxcarbazepine-induced definite cases of DRESS syndrome are rare and oxcarbazepine-induced DRESS syndrome presenting as exfoliative dermatitis is even rarer. We report a case of a 35-year-old male who developed DRESS syndrome presenting as exfoliative dermatitis after taking oxcarbazepine for 3 weeks.

  15. Tire containing thermally exfoliated graphite oxide

    NASA Technical Reports Server (NTRS)

    Prud'homme, Robert K. (Inventor); Aksay, Ilhan A. (Inventor)

    2011-01-01

    A tire, tire lining or inner tube, containing a polymer composite, made of at least one rubber and/or at least one elastomer and a modified graphite oxide material, which is a thermally exfoliated graphite oxide with a surface area of from about 300 sq m/g to 2600 sq m/g.

  16. Psoriasiform exfoliative erythroderma induced by golimumab.

    PubMed

    Mateo, S; García-Martínez, F J; Sánchez-Aguilar, D; Amarelo, J; Toribio, J

    2014-10-01

    Golimumab is a fully human anti-tumour necrosis factor (TNF)-α monoclonal antibody approved for use in the treatment of active rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis. Psoriasis induced by treatment with anti-TNF drugs is well documented, but to our knowledge, the development of clinical features of psoriasiform exfoliative erythroderma during treatment with golimumab has not been previously described.

  17. Production of anti-neurotoxin antibody is enhanced by two subcomponents, HA1 and HA3b, of Clostridium botulinum type B 16S toxin-haemagglutinin.

    PubMed

    Lee, Jae-Chul; Yokota, Kenji; Arimitsu, Hideyuki; Hwang, Hyun-Jung; Sakaguchi, Yoshihiko; Cui, Jinhua; Takeshi, Kouichi; Watanabe, Toshihiro; Ohyama, Tohru; Oguma, Keiji

    2005-11-01

    Clostridium botulinum type B strain produces two forms of progenitor toxin, 16S and 12S. The 12S toxin is formed by association of a neurotoxin (NTX) and a non-toxic non-haemagglutinin (NTNH), and the 16S toxin is formed by conjugation of the 12S toxin with a haemagglutinin (HA). HA consists of four subcomponents designated HA1, HA2, HA3a and HA3b. When mice were immunized with formalin-detoxified NTX, 12S or 16S, a significantly greater amount of anti-NTX antibody (Ab) was produced in the mice injected with 16S than in NTX- or 12S-injected mice. Immunization with NTX mixed with HA1 and/or HA3b also increased the anti-NTX Ab production, whereas NTX mixed with HA2 did not, indicating that HA1 and HA3b have adjuvant activity. This was further confirmed by immunizing mice with human albumin (Alb) alone or Alb mixed with either HA1 or HA3b. When mouse-spleen cells were stimulated with NTX, 16S or different HA subcomponents, 16S, HA1, HA3b and the mixture of HA1 and HA3 significantly increased interleukin 6 (IL6) production compared with NTX alone. Transcription of IL6 mRNA was low after stimulation with NTX alone, but increased to 16S-stimulation levels when NTX was mixed with HA1 or HA3b. In flow cytometry using labelled Abs against CD3 and CD19, the percentage of CD19 cells was higher following stimulation with 16S or NTX mixed with HA1 or HA3b compared with stimulation with NTX. The percentage of CD3 cells remained unchanged. These results suggest strongly that HA1 and HA3b demonstrate adjuvant activity via increasing IL6 production.

  18. The Hcp proteins fused with diverse extended-toxin domains represent a novel pattern of antibacterial effectors in type VI secretion systems.

    PubMed

    Ma, Jiale; Pan, Zihao; Huang, Jinhu; Sun, Min; Lu, Chengping; Yao, Huochun

    2017-01-06

    The type VI secretion system (T6SS) is a widespread molecular weapon deployed by many bacterial species to target eukaryotic host cells or rival bacteria. Using a dynamic injection mechanism, diverse effectors can be delivered by T6SS directly into recipient cells. Here, we report a new family of T6SS effectors encoded by extended Hcps carrying diverse toxin domains. Bioinformatic analyses revealed that these Hcps with C-terminal extension toxins, designated as Hcp-ET, exist widely in the Enterobacteriaceae. To verify our findings, Hcp-ET1 was tested for its antibacterial effect, and showed effective inhibition of target cell growth via the predicted HNH-DNase activity by T6SS-dependent delivery. Further studies showed that Hcp-ET2 mediated interbacterial antagonism via a Tle1 phospholipase (encoded by DUF2235 domain) activity. Notably, comprehensive analyses of protein homology and genomic neighborhoods revealed that Hcp-ET3-4 is fused with 2 toxin domains (Pyocin S3 and Colicin-DNase) C-terminally, and its encoding gene is followed 3 duplications of the cognate immunity genes. However, some bacteria encode a separated hcp-et3 and an orphan et4 (et4O1) genes caused by a termination-codon mutation in the fusion region between Pyocin S3 and Colicin-DNase encoding fragments. Our results demonstrated that both of these toxins had antibacterial effects. Further, all duplications of the cognate immunity protein contributed to neutralize the DNase toxicity of Pyocin S3 and Colicin, which has not been reported previously. In conclusion, we propose that Hcp-ET proteins are polymorphic T6SS effectors, and thus present a novel encoding pattern of T6SS effectors.

  19. Long-Term Effects of Botulinum Toxin Complex Type A Injection on Mechano- and Metabo-Sensitive Afferent Fibers Originating from Gastrocnemius Muscle

    PubMed Central

    Caron, Guillaume; Marqueste, Tanguy; Decherchi, Patrick

    2015-01-01

    The aim of the present study was to investigate long term effects of motor denervation by botulinum toxin complex type A (BoNT/A) from Clostridium Botulinum, on the afferent fibers originating from the gastrocnemius muscle of rats. Animals were divided in 2 experimental groups: 1) untreated animals acting as control and 2) treated animals in which the toxin was injected in the left muscle, the latter being itself divided into 3 subgroups according to their locomotor recovery with the help of a test based on footprint measurements of walking rats: i) no recovery (B0), ii) 50% recovery (B50) and iii) full recovery (B100). Then, muscle properties, metabosensitive afferent fiber responses to potassium chloride (KCl) and lactic acid injections and Electrically-Induced Fatigue (EIF), and mechanosensitive responses to tendon vibrations were measured. At the end of the experiment, rats were killed and the toxin injected muscles were weighted. After toxin injection, we observed a complete paralysis associated to a loss of force to muscle stimulation and a significant muscle atrophy, and a return to baseline when the animals recover. The response to fatigue was only decreased in the B0 group. The responses to KCl injections were only altered in the B100 groups while responses to lactic acid were altered in the 3 injected groups. Finally, our results indicated that neurotoxin altered the biphasic pattern of response of the mechanosensitive fiber to tendon vibrations in the B0 and B50 groups. These results indicated that neurotoxin injection induces muscle afferent activity alterations that persist and even worsen when the muscle has recovered his motor activity. PMID:26485650

  20. Long-Term Effects of Botulinum Toxin Complex Type A Injection on Mechano- and Metabo-Sensitive Afferent Fibers Originating from Gastrocnemius Muscle.

    PubMed

    Caron, Guillaume; Marqueste, Tanguy; Decherchi, Patrick

    2015-01-01

    The aim of the present study was to investigate long term effects of motor denervation by botulinum toxin complex type A (BoNT/A) from Clostridium Botulinum, on the afferent fibers originating from the gastrocnemius muscle of rats. Animals were divided in 2 experimental groups: 1) untreated animals acting as control and 2) treated animals in which the toxin was injected in the left muscle, the latter being itself divided into 3 subgroups according to their locomotor recovery with the help of a test based on footprint measurements of walking rats: i) no recovery (B0), ii) 50% recovery (B50) and iii) full recovery (B100). Then, muscle properties, metabosensitive afferent fiber responses to potassium chloride (KCl) and lactic acid injections and Electrically-Induced Fatigue (EIF), and mechanosensitive responses to tendon vibrations were measured. At the end of the experiment, rats were killed and the toxin injected muscles were weighted. After toxin injection, we observed a complete paralysis associated to a loss of force to muscle stimulation and a significant muscle atrophy, and a return to baseline when the animals recover. The response to fatigue was only decreased in the B0 group. The responses to KCl injections were only altered in the B100 groups while responses to lactic acid were altered in the 3 injected groups. Finally, our results indicated that neurotoxin altered the biphasic pattern of response of the mechanosensitive fiber to tendon vibrations in the B0 and B50 groups. These results indicated that neurotoxin injection induces muscle afferent activity alterations that persist and even worsen when the muscle has recovered his motor activity.

  1. Susceptibility of Anthonomus grandis (cotton boll weevil) and Spodoptera frugiperda (fall armyworm) to a cry1ia-type toxin from a Brazilian Bacillus thuringiensis strain.

    PubMed

    Grossi-de-Sa, Maria Fatima; Quezado de Magalhaes, Mariana; Silva, Marilia Santos; Silva, Shirley Margareth Buffon; Dias, Simoni Campos; Nakasu, Erich Yukio Tempel; Brunetta, Patricia Sanglard Felipe; Oliveira, Gustavo Ramos; Neto, Osmundo Brilhante de Oliveira; Sampaio de Oliveira, Raquel; Soares, Luis Henrique Barros; Ayub, Marco Antonio Zachia; Siqueira, Herbert Alvaro Abreu; Figueira, Edson L Z

    2007-09-30

    Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 microg/mL and 5 microg/mL, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

  2. Bacteria associated with processed crawfish and potential toxin production by Clostridium botulinum type E in vacuum-packaged and aerobically packaged crawfish tails.

    PubMed

    Lyon, W J; Reddmann, C S

    2000-12-01

    Refrigerated vacuum-packaged storage has been shown to increase significantly the shelf life of fresh fish and seafood products, but the effect, if any, on the outgrowth and toxin production of Clostridium botulinum type E on cooked crawfish is unknown. Microflora associated with live crawfish reflect the microbial populations of the harvest water and sediments in which they are living. The presence or absence of specific pathogens in either vacuum-packaged or air-permeable bags of cooked crawfish have not been thoroughly evaluated. This study evaluates the potential survival and outgrowth of biological hazards in both vacuum-packaged and air-permeable-packaged cooked crawfish held at 4 and 10 degrees C for 30 days. During shelf-life studies of vacuum-packaged and air-permeable-bagged cooked crawfish, a total of 31 bacterial species were isolated and identified from crawfish samples using both selective and nonselective media. The only pathogens isolated from both vacuum-packed and air-permeable bags of processed crawfish samples during shelf-life studies were strains of Aeromonas hydrophila and Staphylococcus aureus. C. botulinum type E and Clostridium perfringens species were not isolated from any of the uninoculated crawfish samples. Cooked crawfish were inoculated with 10(3) C. botulinum type E spores per g of crawfish tail meat to determine whether cooked crawfish tails would support the growth of C. botulinum type E strains and produce toxin at refrigerated temperatures. Spore-inoculated crawfish tails were vacuum packaged in both a high barrier film and an air-permeable bag and stored at 4 degrees C and 10 degrees C for 30 days. C. botulinum toxin E was not detected in any of the spore-inoculated packages throughout the shelf-life study until day 30. Microbiological data from this study should be useful in the development and implementation of the hazard analysis and critical control point plans for processed crawfish tails.

  3. Subcutaneous Botulinum toxin type A reduces capsaicin-induced trigeminal pain and vasomotor reactions in human skin.

    PubMed

    Gazerani, Parisa; Pedersen, Natalia Spicina; Staahl, Camilla; Drewes, Asbjørn Mohr; Arendt-Nielsen, Lars

    2009-01-01

    The present human study aimed at investigating the effect of subcutaneous administration of Botulinum toxin type A (BoNT/A) on capsaicin-induced trigeminal pain, neurogenic inflammation and experimentally induced cutaneous pain modalities. Fourteen healthy males (26.3+/-2.6 years) were included in this double-blind and placebo-controlled trial. The subjects received subcutaneous BoNT/A (22.5U) and isotonic saline in the mirror sides of their forehead. Pain and neurogenic inflammation was induced by four intradermal injections of capsaicin (100mug/muL) (before, and days 1, 3 and 7 after treatments). The capsaicin-induced pain intensity, pain area, the area of secondary hyperalgesia, the area of visible flare and vasomotor reactions were recorded together with cutaneous heat, electrical and pressure pain thresholds. BoNT/A reduced the capsaicin-induced trigeminal pain intensity compared to saline (F=37.9, P<0.001). The perceived pain area was smaller for the BoNT/A-treated side compared to saline (F=7.8, P<0.05). BoNT/A reduced the capsaicin-induced secondary hyperalgesia (F=5.3, P<0.05) and flare area (F=10.3, P<0.01) compared to saline. BoNT/A reduced blood flow (F(1,26)=109.5, P<0.001) and skin temperature (F(1,26)=63.1, P<0.001) at the capsaicin injection sites compared to saline and its suppressive effect was maximal at days 3 and 7 (P<0.05, post hoc test). BoNT/A elevated cutaneous heat pain thresholds (F=17.1, P<0.001) compared to saline; however, no alteration was recorded for electrical or pressure pain thresholds (P>0.05). Findings from the present study suggest that BoNT/A appears to preferentially target Cfibers and probably TRPV1-receptors, block neurotransmitter release and subsequently reduce pain, neurogenic inflammation and cutaneous heat pain threshold.

  4. Muscle volume alterations in spastic muscles immediately following botulinum toxin type-A treatment in children with cerebral palsy.

    PubMed

    Williams, Sîan A; Reid, Siobhan; Elliott, Catherine; Shipman, Peter; Valentine, Jane

    2013-09-01

    With evidence for an atrophic effect of botulinum toxin type A (BoNT-A) documented in typically developing muscles, this study investigated the immediate morphological alterations of muscles in children with cerebral palsy (CP) after BoNT-A treatment. Fifteen children (10 males, five females; age range 5-11y, mean age 8y 5mo, SD 1y 10mo) with spastic diplegic CP [Gross Motor Function Classification System Levels I (n=9) and II (n=6)] receiving BoNT-A injections for spasticity management were included. None of the children was a first-time receiver of BoNT-A. Magnetic resonance imaging and Mimics software assessed muscle volume, timed 2 weeks before and 5 weeks after injection. All participants received BoNT-A bilaterally to the gastrocnemius muscle, and five participants also received BoNT-A bilaterally to the medial hamstring muscles. Functional assessment measures used were the 6-Minute Walk Test (6-MWT), the Timed Up and Go (TUG) test, and hand-held dynamometry. Whilst total muscle group volume of the injected muscle group remained unchanged, a 4.47% decrease in the injected gastrocnemius muscle volume (p=0.01) and a 3.96% increase in soleus muscle volume (p=0.02) was evident following BoNT-A. There were no statistically significant changes in function after BoNT-A as assessed by the TUG. There was also no statistically significant change in distance covered in the 6-MWT. Muscle strength, as assessed using hand-held dynamometry was also not statistically different after BoNT-A treatment. Muscle volume decreases were observed in the injected muscle (gastrocnemius), with synergistic muscle hypertrophy that appeared to compensate for this decrement. The 4% to 5% decrease in the volume of BoNT-A injected muscles are not dramatic in comparison to reports in recent animal studies, and are a positive indication for BoNT-A, particularly as it also did not negatively alter function. © 2013 Mac Keith Press.

  5. Receptors of anthrax toxin and cell entry.

    PubMed

    van der Goot, Gisou; Young, John A T

    2009-12-01

    Anthrax toxin-receptor interactions are critical for toxin delivery to the host cell cytoplasm. This review summarizes what is known about the molecular details of the protective antigen (PA) toxin subunit interaction with either the ANTXR1 and ANTXR2 cellular receptors, and how receptor-type can dictate the low pH threshold of PA pore formation. The roles played by cellular factors in regulating the endocytosis of toxin-receptor complexes is also discussed.

  6. Solvent exfoliation of transition metal dichalcogenides: dispersibility of exfoliated nanosheets varies only weakly between compounds.

    PubMed

    Cunningham, Graeme; Lotya, Mustafa; Cucinotta, Clotilde S; Sanvito, Stefano; Bergin, Shane D; Menzel, Robert; Shaffer, Milo S P; Coleman, Jonathan N

    2012-04-24

    We have studied the dispersion and exfoliation of four inorganic layered compounds, WS(2), MoS(2), MoSe(2), and MoTe(2), in a range of organic solvents. The aim was to explore the relationship between the chemical structure of the exfoliated nanosheets and their dispersibility. Sonication of the layered compounds in solvents generally gave few-layer nanosheets with lateral dimensions of a few hundred nanometers. However, the dispersed concentration varied greatly from solvent to solvent. For all four materials, the concentration peaked for solvents with surface energy close to 70 mJ/m(2), implying that all four have surface energy close to this value. Inverse gas chromatography measurements showed MoS(2) and MoSe(2) to have surface energies of ∼75 mJ/m(2), in good agreement with dispersibility measurements. However, this method suggested MoTe(2) to have a considerably larger surface energy (∼120 mJ/m(2)). While surface-energy-based solubility parameters are perhaps more intuitive for two-dimensional materials, Hansen solubility parameters are probably more useful. Our analysis shows the dispersed concentration of all four layered materials to show well-defined peaks when plotted as a function of Hansen's dispersive, polar, and H-bonding solubility parameters. This suggests that we can associate Hansen solubility parameters of δ(D) ∼ 18 MPa(1/2), δ(P) ∼ 8.5 MPa(1/2), and δ(H) ∼ 7 MPa(1/2) with all four types of layered material. Knowledge of these properties allows the estimation of the Flory-Huggins parameter, χ, for each combination of nanosheet and solvent. We found that the dispersed concentration of each material falls exponentially with χ as predicted by solution thermodynamics. This work shows that solution thermodynamics and specifically solubility parameter analysis can be used as a framework to understand the dispersion of two-dimensional materials. Finally, we note that in good solvents, such as cyclohexylpyrrolidone, the dispersions are

  7. Snake venomics of Crotalus tigris: the minimalist toxin arsenal of the deadliest Nearctic rattlesnake venom. Evolutionary Clues for generating a pan-specific antivenom against crotalid type II venoms [corrected].

    PubMed

    Calvete, Juan J; Pérez, Alicia; Lomonte, Bruno; Sánchez, Elda E; Sanz, Libia

    2012-02-03

    We report the proteomic and antivenomic characterization of Crotalus tigris venom. This venom exhibits the highest lethality for mice among rattlesnakes and the simplest toxin proteome reported to date. The venom proteome of C. tigris comprises 7-8 gene products from 6 toxin families; the presynaptic β-neurotoxic heterodimeric PLA(2), Mojave toxin, and two serine proteinases comprise, respectively, 66 and 27% of the C. tigris toxin arsenal, whereas a VEGF-like protein, a CRISP molecule, a medium-sized disintegrin, and 1-2 PIII-SVMPs each represent 0.1-5% of the total venom proteome. This toxin profile really explains the systemic neuro- and myotoxic effects observed in envenomated animals. In addition, we found that venom lethality of C. tigris and other North American rattlesnake type II venoms correlates with the concentration of Mojave toxin A-subunit, supporting the view that the neurotoxic venom phenotype of crotalid type II venoms may be described as a single-allele adaptation. Our data suggest that the evolutionary trend toward neurotoxicity, which has been also reported for the South American rattlesnakes, may have resulted by pedomorphism. The ability of an experimental antivenom to effectively immunodeplete proteins from the type II venoms of C. tigris, Crotalus horridus , Crotalus oreganus helleri, Crotalus scutulatus scutulatus, and Sistrurus catenatus catenatus indicated the feasibility of generating a pan-American anti-Crotalus type II antivenom, suggested by the identification of shared evolutionary trends among South and North American Crotalus species.

  8. Discovery of inhibitors of Shiga toxin type 2 by on-plate generation and screening of a focused compound library.

    PubMed

    Dasgupta, Somnath; Kitov, Pavel I; Sadowska, Joanna M; Bundle, David R

    2014-02-03

    A new microtiter-plate-based method for the rapid generation and evaluation of focused compound libraries was developed and applied to screening ligand analogues for the E. coli Shiga-like toxin Stx2a. The method is general, it mitigates the masking of intrinsic affinity gains by multivalency and enables the discovery of potential hits when starting from ligands that exhibit extremely low affinity with proteins that depend on multivalency for their function.

  9. Endothelial binding of beta toxin to small intestinal mucosal endothelial cells in early stages of experimentally induced Clostridium perfringens type C enteritis in pigs.

    PubMed

    Schumacher, V L; Martel, A; Pasmans, F; Van Immerseel, F; Posthaus, H

    2013-07-01

    Beta toxin (CPB) is known to be an essential virulence factor in the development of lesions of Clostridium perfringens type C enteritis in different animal species. Its target cells and exact mechanism of toxicity have not yet been clearly defined. Here, we evaluate the suitability of a neonatal piglet jejunal loop model to investigate early lesions of C. perfringens type C enteritis. Immunohistochemically, CPB was detected at microvascular endothelial cells in intestinal villi during early and advanced stages of lesions induced by C. perfringens type C. This was first associated with capillary dilatation and subsequently with widespread hemorrhage in affected intestinal segments. CPB was, however, not demonstrated on intestinal epithelial cells. This indicates a tropism of CPB toward endothelial cells and suggests that CPB-induced endothelial damage plays an important role in the early stages of C. perfringens type C enteritis in pigs.

  10. A skincare containing retinol adenosine and hyaluronic acid optimises the benefits from a type A botulinum toxin injection.

    PubMed

    Ascher, Benjamin; Fanchon, Chantal; Kanoun-Copy, Leila; Bouloc, Anne; Benech, Florence

    2012-10-01

    A monocentre double-blind two parallel group clinical study was conducted to assess whether a new skincare regimen containing retinol, adenosine and hyaluronic acid, applied after the injection of botulinum toxin A to the glabellar area, provided a beneficial effect. Standardised photographs acquired using LifeViz cameras and zoomed pictures of the glabella and of the crow's feet areas were analysed with automatic well-defined procedures. Perceived efficacy and tolerance were also analysed by comparison between the two groups. A beneficial effect versus placebo-treated group was proven in the group having topically applied the new skincare regimen for 2 months following botulinum toxin A injection with no touch up after 1 month. 3D image analysis showed more rapid results on D10 and enhanced efficacy on M2. Moreover, a beneficial effect independent of injection was measured in the crow's feet area, and analysis of the self-evaluation questionnaire showed enhanced efficacy perceived by the volunteers. A specially developed skincare regimen applied immediately after botulinum toxin A injection completes the beneficial effect of the injection on the glabellar area and offers clinical benefits in fine lines, wrinkles and smoothness on the whole face.

  11. Botulinum toxin type A injections for the management of muscle tightness following total hip arthroplasty: a case series

    PubMed Central

    Bhave, Anil; Zywiel, Michael G; Ulrich, Slif D; McGrath, Mike S; Seyler, Thorsten M; Marker, David R; Delanois, Ronald E; Mont, Michael A

    2009-01-01

    Background Development of hip adductor, tensor fascia lata, and rectus femoris muscle contractures following total hip arthroplasties are quite common, with some patients failing to improve despite treatment with a variety of non-operative modalities. The purpose of the present study was to describe the use of and patient outcomes of botulinum toxin injections as an adjunctive treatment for muscle tightness following total hip arthroplasty. Methods Ten patients (14 hips) who had hip adductor, abductor, and/or flexor muscle contractures following total arthroplasty and had been refractory to physical therapeutic efforts were treated with injection of botulinum toxin A. Eight limbs received injections into the adductor muscle, 8 limbs received injections into the tensor fascia lata muscle, and 2 limbs received injection into the rectus femoris muscle, followed by intensive physical therapy for 6 weeks. Results At a mean final follow-up of 20 months, all 14 hips had increased range in the affected arc of motion, with a mean improvement of 23 degrees (range, 10 to 45 degrees). Additionally all hips had an improvement in hip scores, with a significant increase in mean score from 74 points (range, 57 to 91 points) prior to injection to a mean of 96 points (range, 93 to 98) at final follow-up. There were no serious treatment-related adverse events. Conclusion Botulinum toxin A injections combined with intensive physical therapy may be considered as a potential treatment modality, especially in difficult cases of muscle tightness that are refractory to standard therapy. PMID:19709429

  12. Botulinum toxin type a injections to salivary glands: combination with single event multilevel chemoneurolysis in 2 children with severe spastic quadriplegic cerebral palsy.

    PubMed

    Kim, Heakyung; Lee, Yung; Weiner, Daniel; Kaye, Robin; Cahill, Anne Marie; Yudkoff, Marc

    2006-01-01

    We describe 2 children with severe spastic quadriplegic cerebral palsy (CP) who have significant drooling and frequent aspiration pneumonia. They underwent simultaneous botulinum toxin type A (BTX-A) injections to salivary glands for drooling and prevention of aspiration pneumonia along with single-event multilevel chemoneurolysis (SEMLC) with BTX-A and 5% phenol for severe diffuse spasticity. There was significant improvement in drooling, frequency of aspiration pneumonia, and spasticity without adverse effect. BTX-A injections into the salivary glands, in addition to SEMLC, for these 2 children with medically complicated severe spastic quadriplegic CP, were safe and highly successful procedures, which improved their health-related quality of life.

  13. The effects of botulinum toxin type A on improvement and dynamic spastic equinus correction in children with cerebral palsy – preliminary results

    PubMed Central

    Dimitrijevic, Lidija; Stankovic, Ivona; Nikolic, Dejan; Radovic-Janosevic, Dragana; Zivanovic, Dragoljub

    2014-01-01

    Introduction We evaluated the effects of botulinum toxin type A (BTA) with physical therapy on dynamic foot equinus correction and higher motor functional outcome in children with spastic type of cerebral palsy (CP). Material and methods Ankle joint active and passive movement, gastrocnemial muscle spasticity levels (Modified Ashworth Scale (MAS)), and higher motor functional status (Gross Motor Function Classification System (GMFCS) and Gross Motor Function Measure (GMFM) (GMFM-D – standing and GMFM-E – walking) were assessed before treatment and 3, 8, 16 weeks and 6 months after BTA administration in 12 children. Results There was a significant improvement of active (initial – (–)13.07 ±5.78; 6 months – (–)10.64 ±4.77; p < 0.001) and passive (initial – 4.21 ±2.29; 6 months – 4.71 ±2.16; p < 0.05) ankle joint foot dorsiflexion. GMFM-D and GMFM-E were significantly higher after 3, 8, 16 weeks (p < 0.001) and GMFM-D after 6 months (p < 0.001). Conclusions Botulinum toxin type A administration and physical therapy in patients with spastic CP improves the motion range of dynamic foot equinus after 3 weeks and higher motor functional outcome (standing and walking). PMID:25395950

  14. Liquid-phase exfoliation of flaky graphite

    NASA Astrophysics Data System (ADS)

    Pavlova, Alexandra S.; Obraztsova, Ekaterina A.; Belkin, Alexey V.; Monat, Christelle; Rojo-Romeo, Pedro; Obraztsova, Elena D.

    2016-01-01

    The majority of currently available methods of graphene production have certain drawbacks limiting its scaling. Unlike the others, liquid-phase exfoliation of graphite is a promising technique for high-yield graphene production. In this work, we present our results on one- to four-layer graphene production using various solvents and surfactants from flaky graphite. We suppose that the initial graphite in the form of millimeter-size flakes can be more advantageous for extended graphene flake acquisition than graphite powder consisting of tiny particles used in previous works. Half-centimeter-size graphene films were obtained by depositing exfoliated flakes on an arbitrary substrate. Such films can be useful for electronic and photonic applications.

  15. Characterization of exfoliated/delamination kaolinite

    SciTech Connect

    Sun, Dewen; Li, Bin; Li, Yanfeng; Yu, Cui; Zhang, Bo; Fei, Huafeng

    2011-01-15

    A novel and facile approach for the preparation of exfoliated/delamination kaolinite was reported in this study. Kaolinite was mechanochemically activated by grinding with dimethylsulfoxide in a globe mill for different periods of time, and then the activated samples were treated for several hours at 120 {sup o}C to obtain the precursors of kaolinite. The resulting materials were characterized by X-ray powder diffraction, scanning electron microscopy, transmission electron microscopy, and X-ray photoelectron spectroscopy. The experimental data indicated that the clay layers were well exfoliated/delamination under mechanochemical effect in a significantly short intercalation time. The expansion of the basal spacing (d{sub 001}) of raw kaolinite by 0.40 nm pointed out that the hydrogen bonds between adjacent kaolinite layers were partially broken as a result of the intercalation with dimethylsulfoxide.

  16. AvrRxo1 Is a Bifunctional Type III Secreted Effector and Toxin-Antitoxin System Component with Homologs in Diverse Environmental Contexts

    PubMed Central

    Triplett, Lindsay R.; Shidore, Teja; Long, John; Miao, Jiamin; Wu, Shuchi; Han, Qian; Zhou, Changhe; Ishihara, Hiromichi; Li, Jianyong; Zhao, Bingyu; Leach, Jan E.

    2016-01-01

    Toxin-antitoxin (TA) systems are ubiquitous bacterial systems that may function in genome maintenance and metabolic stress management, but are also thought to play a role in virulence by helping pathogens survive stress. We previously demonstrated that the Xanthomonas oryzae pv. oryzicola protein AvrRxo1 is a type III-secreted virulence factor that has structural similarities to the zeta family of TA toxins, and is toxic to plants and bacteria in the absence of its predicted chaperone Arc1. In this work, we confirm that AvrRxo1 and its binding partner Arc1 function as a TA system when expressed in Escherichia coli. Sequences of avrRxo1 homologs were culled from published and newly generated phytopathogen genomes, revealing that avrRxo1:arc1 modules are rare or frequently inactivated in some species and highly conserved in others. Cloning and functional analysis of avrRxo1 from Acidovorax avenae, A. citrulli, Burkholderia andropogonis, Xanthomonas translucens, and Xanthomonas euvesicatoria showed that some AvrRxo1 homologs share the bacteriostatic and Rxo1-mediated cell death triggering activities of AvrRxo1 from X. oryzae. Additional distant putative homologs of avrRxo1 and arc1 were identified in genomic or metagenomic sequence of environmental bacteria with no known pathogenic role. One of these distant homologs was cloned from the filamentous soil bacterium Cystobacter fuscus. avrRxo1 from C. fuscus caused watersoaking and triggered Rxo1-dependent cell collapse in Nicotiana benthamiana, but no growth suppression in E. coli was observed. This work confirms that a type III effector can function as a TA system toxin, and illustrates the potential of microbiome data to reveal new environmental origins or reservoirs of pathogen virulence factors. PMID:27391081

  17. Raman spectroscopy and oral exfoliative cytology

    NASA Astrophysics Data System (ADS)

    Sahu, Aditi; Shah, Nupur; Mahimkar, Manoj; Garud, Mandavi; Pagare, Sandeep; Nair, Sudhir; Krishna, C. Murali

    2014-03-01

    Early detection of oral cancers can substantially improve disease-free survival rates. Ex vivo and in vivo Raman spectroscopic (RS) studies on oral cancer have demonstrated the applicability of RS in identifying not only malignant and premalignant conditions but also cancer-field-effects: the earliest events in oral carcinogenesis. RS has also been explored for cervical exfoliated cells analysis. Exfoliated cells are associated with several advantages like non-invasive sampling, higher patient compliance, transportation and analysis at a central facility: obviating need for on-site instrumentation. Thus, oral exfoliative cytology coupled with RS may serve as a useful adjunct for oral cancer screening. In this study, exfoliated cells from healthy controls with and without tobacco habits, premalignant lesions (leukoplakia and tobacco-pouch-keratosis) and their contralateral mucosa were collected using a Cytobrush. Cells were harvested by vortexing and centrifugation at 6000 rpm. The cellular yield was ascertained using Neubauer's chamber. Cell pellets were placed on a CaF2 window and Raman spectra were acquired using a Raman microprobe (40X objective) coupled HE-785 Raman spectrometer. Approximately 7 spectra were recorded from each pellet, following which pellet was smeared onto a glass slide, fixed in 95% ethanol and subjected to Pap staining for cytological diagnosis (gold standard). Preliminary PC-LDA followed by leave-one-out cross validation indicate delineation of cells from healthy and all pathological conditions. A tendency of classification was also seen between cells from contralateral, healthy tobacco and site of premalignant lesions. These results will be validated by cytological findings, which will serve as the basis for building standard models of each condition.

  18. Anisotropic Thermal Conductivity of Exfoliated Black Phosphorus.

    PubMed

    Jang, Hyejin; Wood, Joshua D; Ryder, Christopher R; Hersam, Mark C; Cahill, David G

    2015-12-22

    The anisotropic thermal conductivity of passivated black phosphorus (BP), a reactive two-dimensional material with strong in-plane anisotropy, is ascertained. The room-temperature thermal conductivity for three crystalline axes of exfoliated BP is measured by time-domain thermo-reflectance. The thermal conductivity along the zigzag direction is ≈2.5 times higher than that of the armchair direction.

  19. Conductive composites based on exfoliated graphite

    SciTech Connect

    Afanasov, I.M.; Morozov, V.A.; Seleznev, A.N.; Avdeev, V.V.

    2008-06-15

    Conductive composites of exfoliated graphite (EG) and coal-tar pitch have been prepared by mixing the components. The electrical properties of the composites have been studied, and the results have been interpreted in terms of the percolation theory. The threshold EG content for electrical conduction is determined to be similar or equal to 1.5 wt %, independent of the properties of the pitch and EG.

  20. ESBL-plasmids carrying toxin-antitoxin systems can be “cured” of wild-type Escherichia coli using a heat technique

    PubMed Central

    2013-01-01

    Background Plasmid-encoded extended-spectrum beta-lactamase (ESBL)-enzymes are frequently produced by Escherichia coli. Several ESBL-plasmids contain genes for toxin-antitoxin (TA) systems, which assure the maintenance of plasmids in bacteria and prevent the cells from “post-segregational killing”. These systems limit options to “cure” plasmids of ESBL-wild-type strains due to the death of the bacterial cells. A helpful tool to understand the role of ESBL-plasmids in the dissemination of pandemic multi-resistant E. coli are ESBL-plasmid-“cured”-variants (PCVs) and their comparison to ESBL-wild-type strains. The purpose of this study was to construct PCVs of ESBL-wild-type E. coli strains despite the presence of genes for TA systems. Findings Using enhanced temperatures and brain-heart-infusion broth it was possible to construct viable PCVs of wild-type ESBL-E. coli strains. The occurrence of TA system-genes including hok/sok, srnB/C, vagC/D, pemI/K on ESBL-plasmids of replicon types FIA or FIB was demonstrated by bioinformatic analyses. The loss of the plasmid and the genetic identity of PCV and corresponding wild-type strain was confirmed via different methods including plasmid-profile-analysis, pulsed-field gel electrophoresis and bioinformatics using generated whole genome data of the strains. Conclusions This short report describes the successful construction of viable PCVs of ESBL-wild-type E. coli strains. The results are hence surprising due to the fact that all “cured” ESBL-plasmids contained at least one complete toxin-antitoxin system, whose loss would normally mean the death of bacterial cells. PMID:24245987

  1. Transcriptional Profiling of Type II Toxin-Antitoxin Genes of Helicobacter pylori under Different Environmental Conditions: Identification of HP0967-HP0968 System.

    PubMed

    Cárdenas-Mondragón, María G; Ares, Miguel A; Panunzi, Leonardo G; Pacheco, Sabino; Camorlinga-Ponce, Margarita; Girón, Jorge A; Torres, Javier; De la Cruz, Miguel A

    2016-01-01

    Helicobacter pylori is a Gram-negative bacterium that colonizes the human gastric mucosa and is responsible for causing peptic ulcers and gastric carcinoma. The expression of virulence factors allows the persistence of H. pylori in the stomach, which results in a chronic, sometimes uncontrolled inflammatory response. Type II toxin-antitoxin (TA) systems have emerged as important virulence factors in many pathogenic bacteria. Three type II TA systems have previously been identified in the genome of H. pylori 26695: HP0315-HP0316, HP0892-HP0893, and HP0894-HP0895. Here we characterized a heretofore undescribed type II TA system in H. pylori, HP0967-HP0968, which is encoded by the bicistronic operon hp0968-hp0967 and belongs to the Vap family. The predicted HP0967 protein is a toxin with ribonuclease activity whereas HP0968 is an antitoxin that binds to its own regulatory region. We found that all type II TA systems were expressed in H. pylori during early stationary growth phase, and differentially expressed in the presence of urea, nickel, and iron, although, the hp0968-hp0967 pair was the most affected under these environmental conditions. Transcription of hp0968-hp0967 was strongly induced in a mature H. pylori biofilm and when the bacteria interacted with AGS epithelial cells. Kanamycin and chloramphenicol considerably boosted transcription levels of all the four type II TA systems. The hp0968-hp0967 TA system was the most frequent among 317 H. pylori strains isolated from all over the world. This study is the first report on the transcription of type II TA genes in H. pylori under different environmental conditions. Our data show that the HP0967 and HP0968 proteins constitute a bona fide type II TA system in H. pylori, whose expression is regulated by environmental cues, which are relevant in the context of infection of the human gastric mucosa.

  2. Mass spectrometry-based method of detecting and distinguishing type 1 and type 2 Shiga-like toxins in human serum

    USDA-ARS?s Scientific Manuscript database

    Shiga-like toxins (verotoxins) are a class of AB5 holotoxins that are responsible for the virulence associated with bacterial pathogens such as Shigella dysenteriae, shigatoxigenic and enterohemorrhagic strains of Escherichia coli (STEC and EHEC), and some Enterobacter strains. The actual expression...

  3. Fabrication of Boron Nitride Nanosheets by Exfoliation.

    PubMed

    Wang, Zifeng; Tang, Zijie; Xue, Qi; Huang, Yan; Huang, Yang; Zhu, Minshen; Pei, Zengxia; Li, Hongfei; Jiang, Hongbo; Fu, Chenxi; Zhi, Chunyi

    2016-06-01

    Nanomaterials with layered structures, with their intriguing properties, are of great research interest nowadays. As one of the primary two-dimensional nanomaterials, the hexagonal boron nitride nanosheet (BNNS, also called white graphene), which is an analogue of graphene, possesses various attractive properties, such as high intrinsic thermal conductivity, excellent chemical and thermal stability, and electrical insulation properties. After being discovered, it has been one of the most intensively studied two-dimensional non-carbon nanomaterials and has been applied in a wide range of applications. To support the exploration of applications of BNNSs, exfoliation, as one of the most promising approaches to realize large-scale production of BNNSs, has been intensively investigated. In this review, methods to yield BNNSs by exfoliation will be summarized and compared with other potential fabrication methods of BNNSs. In addition, the future prospects of the exfoliation of h-BN will also be discussed. © 2016 The Chemical Society of Japan & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Effect of Clostridium botulinum toxin type A injections into the deep digital flexor muscle on the range of motion of the metacarpus and carpus, and the force distribution underneath the hooves, of sound horses at the walk.

    PubMed

    Hardeman, Lotte C; van der Meij, Bram R; Oosterlinck, Maarten; Veraa, Stefanie; van der Kolk, Johannes H; Wijnberg, Inge D; Back, Willem

    2013-12-01

    In the treatment of laminitis, reducing deep digital flexor muscle (DDFM) activity might diminish its pull on the distal phalanx, thereby preventing displacement and providing pain relief. Injection of Clostridium botulinum toxin type A into the DDFM of horses is potentially therapeutic. However, the effects of C. botulinum toxin type A on the gait characteristics of sound horses at the walk are not known. The aim of this study was to test if a reduced DDFM activity would lead to (1) alterations of the sagittal range of motion of the metacarpus (SROM) and range of motion of the carpal joint (CROM); (2) changes in the force distribution underneath the hoof (toe vs. heel region: balance index); and (3) changes in the force distribution between the treated and untreated limb (symmetry index). The DDFMs of the left forelimbs of seven sound Royal Dutch Sport Horses were injected with 200 IU C. botulinum toxin type A using electromyography and ultrasound guidance. Measurements using an inertial sensor system and dynamically calibrated pressure plate were performed before and after injections. The SROM and CROM of the treated limb were significantly increased after C. botulinum toxin type A injections. No significant changes were detected in the balance index or in the symmetry index, indicating that no lameness was induced. C. botulinum toxin type A injections into the DDFM of sound horses do not appear to result in substantial gait alterations at the walk. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Consensus panel's assessment and recommendations on the use of 3 botulinum toxin type A products in facial aesthetics.

    PubMed

    Lorenc, Z Paul; Kenkel, Jeffrey M; Fagien, Steven; Hirmand, Haideh; Nestor, Mark S; Sclafani, Anthony P; Sykes, Jonathan M; Waldorf, Heidi A

    2013-03-01

    In this summary article, the authors discuss the characteristics of abobotulinumtoxinA, incobotulinumtoxinA, and onabotulinumtoxinA. With 3 neuromodulators available in the US market, comparisons between and among products will invariably be made, so arguments for the most effective facial aesthetic uses of each neuromodulator are presented. Topics addressed in this article include patient expectations, toxin reconstitution and preparation, patient positioning, differences among products, the role of complexing proteins, and dosing and injection strategies. Recommendations are also provided by treatment area.

  6. Toxin Levels in Serum Correlate with the Development of Staphylococcal Scalded Skin Syndrome in a Murine Model

    PubMed Central

    Plano, Lisa R. W.; Adkins, Becky; Woischnik, Markus; Ewing, Ruth; Collins, Carleen M.

    2001-01-01

    Staphylococcal scalded skin syndrome (SSSS) is an exfoliative dermatitis that results from infection with exfoliative toxin-producing Staphylococcus aureus. SSSS is seen primarily in infants and children. Here we ask if there is a specific maturation process that protects healthy adults from this syndrome. For these studies, an active recombinant exfoliative toxin A (rETA) was used in a neonatal mouse model. A time course generated on the susceptibility to the toxin as a function of mouse age indicated that BALB/c mice developed the characteristic symptoms of SSSS until day 7 of life. Between day 7 and day 8 of life there was a dramatic decrease in susceptibility, such that mice at day 9 of life were resistant to the effects of the toxin. This time course corresponds approximately to the time needed for maturation of the adaptive immune response, and SSSS in adults is often identified with immunocompromised states. Therefore, mice deficient in this response were examined. Adult mice thymectomized at birth and adult SCID mice did not develop the symptoms of SSSS after injection with the toxin, indicating that the adaptive immune response is not responsible for the lack of susceptibility observed in the older mice. SSSS in adults is also associated with renal disorders, suggesting that levels of toxin in serum are important in the development of the disease. rETA was not cleared as efficiently from the serum of 1-day-old mice compared to clearance from 10-day-old mice. Ten-day-old mice were given repeated injections of toxin so that the maximal level of toxin was maintained for a sustained period of time, and exfoliation occurred in these mice. Thus, whereas the adaptive immune response is not needed for protection of adult mice from SSSS, efficient clearance of the toxin from the bloodstream is a critical factor. PMID:11447206

  7. Intracerebroventricular administration of Shiga toxin type 2 induces striatal neuronal death and glial alterations: an ultrastructural study.

    PubMed

    Goldstein, Jorge; Loidl, César Fabián; Creydt, Virginia Pistone; Boccoli, Javier; Ibarra, Cristina

    2007-08-03

    Shiga toxin (Stx) from enterohemorrhagic Escherichia coli (STEC) is the main cause of hemorrhagic colitis which may derive to hemolytic-uremic syndrome (HUS). HUS is characterized by acute renal failure, thrombocytopenia and microangiopathic hemolytic anemia. Mortality in the acute stage has been lower than 5% of total affected argentine children with endemic HUS. Common signs of severe CNS involvement leading to death included seizures, alteration of consciousness, hemiparesis, visual disturbances, and brainstem symptoms. The main purpose of the present work was to study the direct involvement of Stx2 in brain cells by intracerebroventricular (i.c.v.) administration of Stx2. Immunodetection of Stx2 was confirmed by immunoelectron cytochemistry in different subsets and compartments of affected caudate putamen cells of corpus striatum. Transmission electron microscopy (TEM) studies revealed apoptotic neurons, glial ultrastructural alterations and demyelinated fibers. The i.c.v. microinfusion was applied for the first time in rats to demonstrate the direct action of Stx2 in neurons and glial cells. The toxin may affect brain neuroglial cells without the involvement of proinflammatory or systemic neurotoxic elements.

  8. Porous calcium niobate nanosheets prepared by an exfoliation-restacking route.

    PubMed

    Hashemzadeh, Fatemeh

    2016-01-01

    The single phase layered perovskite-type niobate KCa2Nb3O10 was obtained by a solid state reaction of the starting materials (K2CO3, CaCO3 and Nb2O5) at 1,200 °C. Then the H(+)-exchanged form (HCa2Nb3O10) was successfully exfoliated into colloidal porous single layers on the intercalating action of tetra(butyl)ammonium ion. The various characterization techniques such as X-ray diffraction (XRD), field-emission scanning electron microscopy, N2 absorption-desorption and diffuse reflectance UV-visible spectrometry gave important information on the unusual structural features of the perovskite-related niobate nanosheets. XRD analysis of the exfoliated nanosheets showed a unique profile with wide peaks that represented individual molecular aspects of the nanosheets. The Brunauer-Emmett-Teller isotherm of the exfoliated coiled nanosheets showed a sharp increase in the surface area by a factor of >30 in comparison to parent layered material, which is due to the exfoliation and restacking process. The nanosheets in this study were also found to act as a semiconductor with a wide band gap that is due to the quantum size effect.

  9. Toxins of filamentous fungi.

    PubMed

    Bhatnagar, Deepak; Yu, Jiujiang; Ehrlich, Kenneth C

    2002-01-01

    Mycotoxins are low-molecular-weight secondary metabolites of fungi. The most significant mycotoxins are contaminants of agricultural commodities, foods and feeds. Fungi that produce these toxins do so both prior to harvest and during storage. Although contamination of commodities by toxigenic fungi occurs frequently in areas with a hot and humid climate (i.e. conditions favorable for fungal growth), they can also be found in temperate conditions. Production of mycotoxins is dependent upon the type of producing fungus and environmental conditions such as the substrate, water activity (moisture and relative humidity), duration of exposure to stress conditions and microbial, insect or other animal interactions. Although outbreaks of mycotoxicoses in humans have been documented, several of these have not been well characterized, neither has a direct correlation between the mycotoxin and resulting toxic effect been well established in vivo. Even though the specific modes of action of most of the toxins are not well established, acute and chronic effects in prokaryotic and eukaryotic systems, including humans have been reported. The toxicity of the mycotoxins varies considerably with the toxin, the animal species exposed to it, and the extent of exposure, age and nutritional status. Most of the toxic effects of mycotoxins are limited to specific organs, but several mycotoxins affect many organs. Induction of cancer by some mycotoxins is a major concern as a chronic effect of these toxins. It is nearly impossible to eliminate mycotoxins from the foods and feed in spite of the regulatory efforts at the national and international levels to remove the contaminated commodities. This is because mycotoxins are highly stable compounds, the producing fungi are ubiquitous, and food contamination can occur both before and after harvest. Nevertheless, good farm management practices and adequate storage facilities minimize the toxin contamination problems. Current research is

  10. Genotoxic and histopathological biomarkers for assessing the effects of magnetic exfoliated vermiculite and exfoliated vermiculite in Danio rerio.

    PubMed

    Cáceres-Vélez, Paolin Rocio; Fascineli, Maria Luiza; Grisolia, Cesar Koppe; de Oliveira Lima, Emília Celma; Sousa, Marcelo Henrique; de Morais, Paulo César; Bentes de Azevedo, Ricardo

    2016-05-01

    Magnetic exfoliated vermiculite is a synthetic nanocomposite that quickly and efficiently absorbs organic compounds such as oil from water bodies. It was developed primarily to mitigate pollution, but the possible adverse impacts of its application have not yet been evaluated. In this context, the acute toxicity of magnetic exfoliated vermiculite and exfoliated vermiculite was herein assessed by genotoxic and histopathological biomarkers in zebrafish (Danio rerio). DNA fragmentation was statistically significant for all groups exposed to the magnetic exfoliated vermiculite and for fish exposed to the highest concentration (200mg/L) of exfoliated vermiculite, whereas the micronucleus frequency, nuclear abnormalities and histopathological alterations were not statistically significant for the fish exposed to these materials. In the intestinal lumen, epithelial cells and goblet cells, we found the presence of magnetic exfoliated vermiculite and exfoliated vermiculite, but no alterations or presence of the materials-test in the gills or liver were observed. Our findings suggest that the use of magnetic exfoliated vermiculite and exfoliated vermiculite during standard ecotoxicological assays caused DNA damage in D. rerio, whose alterations may be likely to be repaired, indicating that the magnetic nanoparticles have the ability to promote genotoxic damage, such as DNA fragmentation, but not mutagenic effects.

  11. Revised nomenclature of Clostridium difficile toxins and associated genes.

    PubMed

    Rupnik, Maja; Dupuy, Bruno; Fairweather, Neil F; Gerding, Dale N; Johnson, Stuart; Just, Ingo; Lyerly, David M; Popoff, Michel R; Rood, Julian I; Sonenshein, Abraham L; Thelestam, Monica; Wren, Brendan W; Wilkins, Tracy D; von Eichel-Streiber, Christoph

    2005-02-01

    Several different nomenclatures have been applied to the Clostridium difficile toxins and their associated genes. This paper summarizes the new nomenclature that has been agreed to by the research groups currently active in the field. The revised nomenclature includes C. difficile toxins and other related large clostridial toxins produced by Clostridium sordellii and Clostridium novyi, and corresponding toxin genes, as well as toxin production types of C. difficile strains.

  12. Effect of post-exfoliation treatments on mechanically exfoliated MoS2

    NASA Astrophysics Data System (ADS)

    Budania, P.; Baine, P. T.; Montgomery, J. H.; McNeill, D. W.; Mitchell, S. J. N.; Modreanu, M.; Hurley, P. K.

    2017-02-01

    Post-exfoliation thermal annealing in air and ultrasonic treatments were carried out on mechanically exfoliated MoS2 flakes on oxidized silicon substrates. Ultra-sonication of MoS2 flakes on SiO2 without prior annealing results in almost complete removal of flakes, indicating weak interface bonding. The interface adhesion between MoS2 flakes and the substrate is significantly improved when the samples are annealed at 270 °C as the flakes remain strongly adhered to the substrate during subsequent ultrasonic treatment. We consider that improved adhesion is due to greater contact area between the flakes and the substrate due to effusion of trapped impurities during annealing. Annealing between 75 °C and 175 °C followed by ultrasonic treatment results in small MoS2 fragments on the samples due to breakage and/or partial removal of top layers. It also results in exposing residual adhesive traces on the sample which are caught between the flake and the substrate during repetitive folding of the Scotch® tape during the initial exfoliation. An annealing temperature of 460 °C results in decomposition of MoS2 and formation of MoO3. Optical microscopy, non-contact-mode atomic force microscopy (AFM) and Raman spectroscopy were used for identification of MoS2 fragments and residual traces left on the samples after the post-exfoliation treatments.

  13. Botulinum Toxin for Rhinitis.

    PubMed

    Ozcan, Cengiz; Ismi, Onur

    2016-08-01

    Rhinitis is a common clinical entity. Besides nasal obstruction, itching, and sneezing, one of the most important symptoms of rhinitis is nasal hypersecretion produced by nasal glands and exudate from the nasal vascular bed. Allergic rhinitis is an IgE-mediated inflammatory reaction of nasal mucosa after exposure to environmental allergens. Idiopathic rhinitis describes rhinitis symptoms that occur after non-allergic, noninfectious irritants. Specific allergen avoidance, topical nasal decongestants, nasal corticosteroids, immunotherapy, and sinonasal surgery are the main treatment options. Because the current treatment modalities are not enough for reducing rhinorrhea in some patients, novel treatment options are required to solve this problem. Botulinum toxin is an exotoxin generated by Clostridium botulinum. It disturbs the signal transmission at the neuromuscular and neuroglandular junction by inhibiting the acetylcholine release from the presynaptic nerve terminal. It has been widely used in neuromuscular, hypersecretory, and autonomic nerve system disorders. There have been a lot of published articles concerning the effect of this toxin on rhinitis symptoms. Based on the results of these reports, intranasal botulinum toxin A administration appears to be a safe and effective treatment method for decreasing rhinitis symptoms in rhinitis patients with a long-lasting effect. Botulinum toxin type A will be a good treatment option for the chronic rhinitis patients who are resistant to other treatment methods.

  14. Exfoliation syndrome: assembling the puzzle pieces.

    PubMed

    Pasquale, Louis R; Borrás, Terete; Fingert, John H; Wiggs, Janey L; Ritch, Robert

    2016-09-01

    To summarize various topics and the cutting edge approaches to refine XFS pathogenesis that were discussed at the 21st annual Glaucoma Foundation Think Tank meeting in New York City, Sept. 19-20, 2014. The highlights of three categories of talks on cutting edge research in the field were summarized. Exfoliation syndrome (XFS) is a systemic disorder with a substantial ocular burden, including high rates of cataract, cataract surgery complications, glaucoma and retinal vein occlusion. New information about XFS is akin to puzzle pieces that do not quite join together to reveal a clear picture regarding how exfoliation material (XFM) forms. Meeting participants concluded that it is unclear how the mild homocysteinemia seen in XFS might contribute to the disarrayed extracellular aggregates characteristic of this syndrome. Lysyl oxidase-like 1 (LOXL1) variants are unequivocally genetic risk factors for XFS but exactly how these variants contribute to the assembly of exfoliation material (XFM) remains unclear. Variants in a new genomic region, CACNA1A associated with XFS, may alter calcium concentrations at the cell surface and facilitate XFM formation but much more work is needed before we can place this new finding in proper context. It is hoped that various animal model and ex vivo systems will emerge that will allow for proper assembly of the puzzle pieces into a coherent picture of XFS pathogenesis. A clear understanding of XFS pathogenesis may lead to 'upstream solutions' to reduce the ocular morbidity produced by XFS. © 2015 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  15. Legionella Toxin.

    DTIC Science & Technology

    1981-04-29

    of a cytotoxin produced by Legionella pneumophila. Infect. Immun. 29:271-274. Fumarola, D. (1978) Legionnaires ’ disease agent and Limulus endotoxin... Legionnaires ’ disease bacterium in the AKR/J mouse. Ann. intern. Med. 90:676-679. Hedlund, K. W. and Larson, R. (1981) Legionella pneumophila toxin, isolation... Legionella Species New Name Old Name Lpneumophila Legionnaires ’ disease organism, OLDA L. bozemanil WIGA, MI 15 L. dumoffii NY 23, TEX-KL *L. micdadei

  16. Adhesive taping vs. daily manual muscle stretching and splinting after botulinum toxin type A injection for wrist and fingers spastic overactivity in stroke patients: a randomized controlled trial.

    PubMed

    Santamato, Andrea; Micello, Maria Francesca; Panza, Francesco; Fortunato, Francesca; Picelli, Alessandro; Smania, Nicola; Logroscino, Giancarlo; Fiore, Pietro; Ranieri, Maurizio

    2015-01-01

    To compare the effectiveness of two procedures increasing the botulinum toxin type A effect for wrist and finger flexor spasticity after stroke. A single-blind randomized trial. Seventy patients with upper limb post-stroke spasticity. Adults with wrist and finger flexor muscles spasticity after stroke were submitted to botulinum toxin type A therapy. After the treatment, the subjects injected were randomly divided into two groups and submitted to adhesive taping (Group A) or daily muscle manual stretching, passive articular mobilization of wrist and fingers, and palmar splint (Group B) for 10 days. We measured spasticity with Modified Ashworth Scale, related disability with Disability Assessment Scale, and fingers position at rest. The measurements were done at baseline, after two weeks, and after one month from the treatment session. After two weeks, subjects in Group A reported a significantly greater decrease in spasticity scores (Modified Ashworth Scale fingers: mean (standard deviation) 1.3±0.6 vs. 2.1±0.6; Modified Ashworth Scale wrist: 1.7 ±0.6 vs. 2.3 ±0.8), and after one month in spasticity and disability scores (Modified Ashworth Scale fingers: mean (standard deviation) 1.9 ±0.7 vs. 2.5 ±0.6; Modified Ashworth Scale wrist: 2.0 ±0.7 vs. 2.6 ±0.6; Disability Assessment Scale: 1.6 ±0.7 vs. 2.1 ±0.7) compared with Group B subjects. Subjects in Group A reported also a significantly improved fingers position at rest compared with Group B subjects after two weeks (2.8 ±0.9 vs. 2.1 ±0.7) and one month (2.3 ±0.7 vs. 1.5 ±0.6). Adhesive taping of wrist and finger flexor muscles appeared to enhance the effect of botulinum toxin type A therapy more than daily manual muscle stretching combined with passive articular mobilization and palmar splint. © The Author(s) 2014.

  17. Doping of graphene during chemical exfoliation

    NASA Astrophysics Data System (ADS)

    Srivastava, Pawan Kumar; Yadav, Premlata; Ghosh, Subhasis

    2013-02-01

    Graphene provides a perfect platform to explore the unique electronic properties in two-dimensions. However, most electronic applications are handicapped by the absence of a semiconducting gap in pristine graphene. To control the semiconducting properties of graphene, doping is regarded as one of the most feasible methods. Here we demonstrate that graphene can be effectively doped during chemical exfoliation of highly ordered pyrolitic graphite in organic solvents. Layered structure of graphene sheets was confirmed by confocal Raman spectroscopy and doping was probed by analyzing shift in Raman peak positions and transistor transfer (IDS-VGS) characteristics.

  18. A Novel Pore-Forming Toxin in Type A Clostridium perfringens Is Associated with Both Fatal Canine Hemorrhagic Gastroenteritis and Fatal Foal Necrotizing Enterocolitis

    PubMed Central

    Nowell, Victoria J.; Nicholson, Vivian M.; Oliphant, Kaitlyn; Prescott, John F.

    2015-01-01

    A role for type A Clostridium perfringens in acute hemorrhagic and necrotizing gastroenteritis in dogs and in necrotizing enterocolitis of neonatal foals has long been suspected but incompletely characterized. The supernatants of an isolate made from a dog and from a foal that died from these diseases were both found to be highly cytotoxic for an equine ovarian (EO) cell line. Partial genome sequencing of the canine isolate revealed three novel putative toxin genes encoding proteins related to the pore-forming Leukocidin/Hemolysin Superfamily; these were designated netE, netF, and netG. netE and netF were located on one large conjugative plasmid, and netG was located with a cpe enterotoxin gene on a second large conjugative plasmid. Mutation and complementation showed that only netF was associated with the cytotoxicity. Although netE and netG were not associated with cytotoxicity, immunoblotting with specific antisera showed these proteins to be expressed in vitro. There was a highly significant association between the presence of netF with type A strains isolated from cases of canine acute hemorrhagic gastroenteritis and foal necrotizing enterocolitis. netE and netF were found in all cytotoxic isolates, as was cpe, but netG was less consistently present. Pulsed-field gel electrophoresis showed that netF-positive isolates belonged to a clonal population; some canine and equine netF-positive isolates were genetically indistinguishable. Equine antisera to recombinant Net proteins showed that only antiserum to rNetF had high supernatant cytotoxin neutralizing activity. The identifica-tion of this novel necrotizing toxin is an important advance in understanding the virulence of type A C. perfringens in specific enteric disease of animals. PMID:25853427

  19. Effects of tcpB Mutations on Biogenesis and Function of the Toxin-Coregulated Pilus, the Type IVb Pilus of Vibrio cholerae.

    PubMed

    Gao, Yang; Hauke, Caitlyn A; Marles, Jarrad M; Taylor, Ronald K

    2016-10-15

    Vibrio cholerae is the etiological agent of the acute intestinal disorder cholera. The toxin-coregulated pilus (TCP), a type IVb pilus, is an essential virulence factor of V. cholerae Recent work has shown that TcpB is a large minor pilin encoded within the tcp operon. TcpB contributes to efficient pilus formation and is essential for all TCP functions. Here, we have initiated a detailed targeted mutagenesis approach to further characterize this salient TCP component. We have identified (thus far) 20 residues of TcpB which affect either the steady-state level of TcpB or alter one or more TCP functions. This study provides a solid framework for further understanding of the complex role of TcpB and will be of use upon determination of the crystal structure of TcpB or related minor pilin orthologs of type IVb pilus systems. Type IV pili, such as the toxin-coregulated pilus (TCP) in V. cholerae, are bacterial appendages that often act as essential virulence factors. Minor pilins, like TcpB, of these pili systems often play integral roles in pilus assembly and function. In this study, we have generated mutations in tcpB to determine residues of importance for TCP stability and function. Combined with a predicted tertiary structure, characterization of these mutants allows us to better understand critical residues in TcpB and the role they may play in the mechanisms underlying minor pilin functions. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  20. Distribution and inferred age of exfoliation joints in the Aar Granite of the central Swiss Alps and relationship to Quaternary landscape evolution

    NASA Astrophysics Data System (ADS)

    Ziegler, Martin; Loew, Simon; Moore, Jeffrey R.

    2013-11-01

    Exfoliation joints are well-known natural fractures limited to near the ground surface. Relatively few details, however, are known about their distribution and age in the Swiss Alps. Exfoliation joints follow the landscape surface at the time of their formation; the age of the associated landscape feature then provides a maximum age of exfoliation joints. While landscape forms can change through time, exfoliation joints preserve elements of former landscape morphologies by their undisturbed orientations. The Grimsel region of the Central Alps is well-suited for analyzing the impact of erosional episodes, and accompanying stress changes, on exfoliation joint formation in granitic rocks. Mapping above and below ground revealed that exfoliation joints are widespread and occur between valley bottoms and mountain crests within glacial (inner and hanging U-shaped trough valleys, glacial cirques, and steep mountain crests) and predominantly fluvial landforms (gently inclined linear slopes above the inner trough valleys, narrow inner-valley gorges, and steep V-shaped side gullies). Based primarily on their geometric properties at the ground surface, three exfoliation joint types were distinguished in our study area: (1) closely spaced (< 1 m) joints oriented distinctly parallel to the present-day ground surface, (2) intermediately spaced (0.6-2 m) joints that are nearly parallel (< 10° difference) to today's mean ground surface at a 10-m scale, and (3) widely spaced (≫ 2 m) joints not parallel to the ground surface. Relating the mapped distribution of exfoliation joint types to identified erosional episodes and landscape features of known and inferred ages, respectively, enables us to distinguish four exfoliation joint generations in the Grimsel area, which most likely formed during the lower Pleistocene (~ 1.5-1 Ma), middle Pleistocene (~ 0.7-0.4 Ma), upper Pleistocene (0.1-0.02 Ma), and Late Glacial/Holocene (< 0.02 Ma). We demonstrate that the most prominent and

  1. Clostridium perfringens Delta Toxin Is Sequence Related to Beta Toxin, NetB, and Staphylococcus Pore-Forming Toxins, but Shows Functional Differences

    PubMed Central

    Manich, Maria; Knapp, Oliver; Gibert, Maryse; Maier, Elke; Jolivet-Reynaud, Colette; Geny, Blandine; Benz, Roland; Popoff, Michel R.

    2008-01-01

    Clostridium perfringens produces numerous toxins, which are responsible for severe diseases in man and animals. Delta toxin is one of the three hemolysins released by a number of C. perfringens type C and possibly type B strains. Delta toxin was characterized to be cytotoxic for cells expressing the ganglioside GM2 in their membrane. Here we report the genetic characterization of Delta toxin and its pore forming activity in lipid bilayers. Delta toxin consists of 318 amino acids, its 28 N-terminal amino acids corresponding to a signal peptide. The secreted Delta toxin (290 amino acids; 32619 Da) is a basic protein (pI 9.1) which shows a significant homology with C. perfringens Beta toxin (43% identity), with C. perfringens NetB (40% identity) and, to a lesser extent, with Staphylococcus aureus alpha toxin and leukotoxins. Recombinant Delta toxin showed a preference for binding to GM2, in contrast to Beta toxin, which did not bind to gangliosides. It is hemolytic for sheep red blood cells and cytotoxic for HeLa cells. In artificial diphytanoyl phosphatidylcholine membranes, Delta and Beta toxin formed channels. Conductance of the channels formed by Delta toxin, with a value of about 100 pS to more than 1 nS in 1 M KCl and a membrane potential of 20 mV, was higher than those formed by Beta toxin and their distribution was broader. The results of zero-current membrane potential measurements and single channel experiments suggest that Delta toxin forms slightly anion-selective channels, whereas the Beta toxin channels showed a preference for cations under the same conditions. C. perfringens Delta toxin shows a significant sequence homolgy with C. perfringens Beta and NetB toxins, as well as with S. aureus alpha hemolysin and leukotoxins, but exhibits different channel properties in lipid bilayers. In contrast to Beta toxin, Delta toxin recognizes GM2 as receptor and forms anion-selective channels. PMID:19018299

  2. Simple Synthesis of Fluorinated Graphene: Thermal Exfoliation of Fluorographite.

    PubMed

    Jankovský, Ondřej; Mazánek, Vlastimil; Klímová, Kateřina; Sedmidubský, David; Kosina, Jiří; Pumera, Martin; Sofer, Zdeněk

    2016-12-05

    Fluorinated graphene can be prepared directly by thermal exfoliation of fluorographite. The exfoliation was performed in a dynamic nitrogen atmosphere at various temperatures and the exfoliation products were analysed in detail by GC-MS. The structure and properties of all prepared fluorinated graphenes with various contents of fluorine were characterized by a number of analytical techniques. The results show both the dependence of fluorine concentration on exfoliation temperature and the suitability of this method for the synthesis of graphene with controlled concentration of fluorine. The high-temperature exfoliated fluorographite exhibits a high heterogeneous electron transfer rate and excellent catalytic properties towards the oxygen reduction reaction. These synthetic procedures can open a simple way for the synthesis of fluorinated graphene-based devices with tailored properties.

  3. Expression of type 8 capsular polysaccharide and production of toxic shock syndrome toxin 1 are associated among vaginal isolates of Staphylococcus aureus.

    PubMed Central

    Lee, J C; Liu, M J; Parsonnet, J; Arbeit, R D

    1990-01-01

    A colony immunoblot method was developed for serotyping the capsular polysaccharides expressed by Staphylococcus aureus isolates. The method was rapid and specific and was performed with either polyclonal or monoclonal antibodies specific for each of the capsule types. S. aureus isolates were obtained from patients with toxic shock syndrome (TSS) or other staphylococcal infections and from asymptomatic women with vaginal colonization. Among the vaginal isolates of S. aureus, expression of the type 8 capsule was significantly (P less than 0.001) more frequent among strains that produced TSS toxin 1 (TSST-1) than it was among TSST-1-negative strains. In contrast, the frequency of type 8 capsule expression was similar among both TSST-1-positive and -negative strains of S. aureus from patients with nonvaginal TSS. When all vaginal and nonvaginal isolates were compared, TSST-1-negative S. aureus strains were equally distributed among the type 5 and 8 and nontypeable capsule groups, whereas TSST-1-positive strains were predominantly capsule type 8. Images PMID:2279990

  4. The profile of patients and current practice of treatment of upper limb muscle spasticity with botulinum toxin type A: an international survey.

    PubMed

    Bakheit, Abdel Magid; Zakine, Benjamin; Maisonobe, Pascal; Aymard, Claire; Fhedoroff, Klemens; Hefter, Harold; Jacinto, Jorge; Jost, Wolfgang H; Molteni, Franco; Stam, Henk; Turner-Stokes, Lynne; Wissel, Jorg

    2010-09-01

    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 countries were studied. Most patients were over 40 years old and had a stroke. Improvement of active function was the most frequent treatment goal in the first 3 months after the onset of upper limb spasticity, but was less common than passive function in the chronic stage. Pain relief was a common goal in both the stages. As a rule, clinicians intended to assess the effectiveness of treatment with impairment level scales. Functional outcome measures seem to be rarely used in clinical practice. The use of these measures should be encouraged to assess whether the reduction in muscle tone translates into functional benefit to patients and their caregivers.

  5. [An electromyographic study on the development of optimal tactics of botulinum toxin type A injections in children with spastic forms of cerebral palsy].

    PubMed

    Kurenkov, A L; Kuzenkova, L M; Burgasova, B I; Petrova, S A; Klochkova, O A; Nikitin, S S; Artemenko, A R; Mamed'iarov, A M

    2013-01-01

    We studied 67 children, aged 2-9 years, with cerebral palsy including 56 children with a spastic form. An electromyographic method was used for the development of optimal tactics of botulinum toxin type A injections in different clinical presentations of spasticity. The best clinical results were obtained in children with the following changes on EMG: 1) the tonic muscle activity in resting state was minimal (<10 microvolts) and had local or regional distribution; 2) the pathological synkinetic activity during voluntary movements was minimal (synergetic activity coefficient for shin muscles was less than 0.45); 3) the disturbance of interactions between synergistic and antagonistic muscles was moderate (reciprocity coefficient was not less than 0.4); 4) EMG amplitude in voluntary muscle contraction should not be less than 150 microvolts. This approach to the treatment allowed to reach higher levels on The Gross Motor Function Classification System in part of children.

  6. Screening for variations in anterior digastric musculature prior to correction of post-traumatic anterior open bite by injection of botulinum toxin type A: a technical note.

    PubMed

    Zdilla, Matthew J

    2015-06-01

    It has recently been reported that long-standing post-traumatic open bite can be successfully corrected with botulinum toxin type A (BTX-A) injection into the anterior belly of the digastric muscle (ABDM). The report documented an individual with bilaterally symmetrical and otherwise unremarkable anterior digastric musculature. However, the existence of variant anterior digastric musculature is common and may complicate the management of anterior open bite with BTX-A injection. Screening for variant ABDM can be accomplished via ultrasound, computed tomography, and magnetic resonance imaging. Screening for variant ABDM should be performed prior to BTX-A injection in order to account for musculature that may exert undesired forces, such as inferolateral deviation, on the anterior mandible in patients with anterior open bite.

  7. Determination of T-2 toxin, HT-2 toxin, and three other type A trichothecenes in layer feed by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS)--comparison of two sample preparation methods.

    PubMed

    Bernhardt, Katrin; Valenta, Hana; Kersten, Susanne; Humpf, Hans-Ulrich; Dänicke, Sven

    2016-05-01

    A sensitive method for the simultaneous determination of T-2 toxin, HT-2 toxin, neosolaniol, T-2 triol, and T-2 tetraol in layer feed using high-performance liquid chromatography coupled to triple quadrupole mass spectrometry in the positive ionization mode (LC-ESI-MS/MS) is described. Two fast and easy clean-up methods-with BondElut Mycotoxin and MycoSep 227 columns, respectively-were tested. The separation of the toxins was conducted on a Pursuit XRs Ultra 2.8 HPLC column using 0.13 mM ammonium acetate as eluent A and methanol as eluent B. Detection of the mycotoxins was carried out in the multiple reaction monitoring (MRM) mode using ammonium adducts as precursor ions. Quantification of all analytes was performed with d3-T-2 toxin as an internal standard. The clean-up method with MycoSep 227 columns gave slightly better results for layer feed compared to the method using BondElut Mycotoxin columns (MycoSep 227: recovery between 50 and 63%, BondElut Mycotoxin: recovery between 32 and 67%) and was therefore chosen as the final method. The limits of detection ranged between 0.9 and 7.5 ng/g depending on the mycotoxin. The method was developed fo