Syftet med studien var att undersöka patienters upplevelse av smärtlindring och livskvalitet efter botoxbehandling i smärtlindrande syfte samt patienternas inställning till Botox. Metod: En kvalitativ intervjustudie med en deskriptiv och explorativ design. Urvalet bestod av sju patienter, sex kvinnor och en man, vilka behandlas men botoxbehandling vid Smärtcentrum på Akademiska sjukhuset i Uppsala. Resultat: Materialet analyserades enligt syftet under tre kategorier: smärtlindring, oro/nedstämdhet samt inställning till Botox. Samtliga informanter har fått en minskad smärta till följd av botoxbehandlingen och upplever att livskvaliteten förbättrats. Trots biverkningar känner sig patienterna nöjda med behandlingen så länge fördelarna är fler än nackdelarna. Ingen av informanterna förutom en kände någon oro/nedstämdhet. Några informanter upplevde en viss oro inför eventuella biverkningar i framtiden. Slutsats: Botox kan vara ett bra behandlingsalternativ för patienter som lider av långvarig smärta. Samtliga studiedeltagare upplever smärtlindring även om inte smärtan försvinner helt. Behandlingen förbättrar patienternas livskvalitet vilken mäts genom upplevelse av rörlighet, hygien, aktivitet, oro/nedstämdhet och smärta. Hälso- och sjukvården har en viktig roll i att behandla sjukdom och främja hälsa och ge patienterna förutsättningar får att nå hälsorelaterad livskvalitet. / The aim of this study is to explore patients'experience of pain relief and quality of life after Botox treatment of pain-relieving purposes and patients' attitudes to Botox. Method: A qualitative interview study with a descriptive and explorative design. The sample consisted of seven patients, six women and one man, which is treated with the treatment of Botox at Center of pain at the University hospital in Uppsala. Results: The material was analyzed in three categories: pain, anxiety/depression, and attitude to Botox. All informants have experienced a reduction in pain as a result of Botox treatment and feel a higher quality of life. Despite the side effects patients feel satisfied with the treatment so long as the advantages outnumber the disadvantages. None of the respondents except one felt no anxiety/depression. Some respondents felt some concern for possible adverse events in the future. Conclusion: Botox may be a good treatment option for patients suffering from chronic pain. All study participants experienced pain relief even if the pain does not disappear completely. The treatment improves patients' quality of life which is measured by the experience of mobility, hygiene, activity, anxiety/depression and pain. The health service has an important role in treating disease and promoting health and providing patients with conditions, to achieve health-related quality of life.

Les neurotoxines botuliques (BoNTs), produites par C. botulinum, sont responsables du botulisme humain et animal. Dans sa forme naturelle, le botulisme résulte le plus souvent d’une absorption des toxines botuliques à partir du tube digestif après ingestion d’aliments contaminés par la toxine et C. botulinum. L’intoxination peut être divisée en 4 grandes étapes : production de toxine par la bactérie, ingestion d’aliments contenant la toxine préformée, passage de la neurotoxine à travers la barrière intestinale et action protéolytique aux terminaisons nerveuses. La régulation de la production des toxines et le passage des neurotoxines botuliques à travers la barrière intestinale sont mal compris. BoNT s’associe à des protéines non toxiques (NAPs) pour former des complexes de différentes tailles. Les gènes codant les BoNTs et NAPs sont regroupés sur le locus botulique et leur expression est contrôlée positivement par le facteur sigma alternatif BoTR/A. La toxinogénèse chez C. botulinum est contrôlée par un réseau complexe de régulateurs incluant au moins 3 systèmes à deux composants (TCS), identifiés pas la méthode d’ARN antisens, qui régulent positivement la production de complexe botulique indépendamment de BoTR/A. D’autre part, l’entrée de BoNT/B dans la barrière intestinale a été suivie à l’aide du fragment HcB marqué en fluorescence dans une anse intestinale ligaturée de souris. Des analyses en microscopie à fluorescence, immunohistochimie et microscopie électronique ont permis de mettre en évidence que HcB transcytose à travers les entérocytes par une voie d’endocytose dépendante de la dynamine. HcB cible les terminaisons nerveuses acétylcholinergiques de la lamina propia des villosités et gagne les neurones acétylcholinergiques et sérotoninergiques de la sous-muqueuse et de la musculeuse en seulement 10 minutes. Une étude in vitro réalisée sur cellules intestinales (m-ICcl2) montre que l’entrée de HcB est dépendante de récepteurs gangliosidiques GD1b/GT1b présents à la surface des cellules mais pas de la synaptotagmine II qui est requise pour l’entrée de BoNT/B dans les cellules neuronales. / Botulinum neurotoxins (BoNTs), produced by C. botulinum, are responsible for animal and human botulism. In its natural form, botulism is mostly acquired after absorption of BoNTs in the digestive tract after ingestion of food contaminated with C. botulinum and its toxins. The intoxination can be divided in 4 major steps: toxin production, ingestion of food contaminated with BoNTs, passage of BoNTs through the intestinal barrier, and proteolytic activity on nerve endings. Regulation of toxin production and passage of BoNTs through the intestinal barrier are poorly understood. BoNT associates with non toxic protein (NAPs) to form complexes of various sizes. The BoNTs and NAPs genes are clustered in the botulinum locus and are positively regulated by an alternative sigma factor BotR/A. Toxinogenesis in C. botulinum is regulated by a complex regulatory network containing at least 3 two components systems (TCS), identified by antisens RNA strategy, which regulate the production of botulinum complex independently of BotR/A. On the other hand, BoNT/B entry was monitored with fluorescent HcB fragment in ligatureted mouse intestinal loop. Fluorescent imaging analysis, immunohistochemistry and electron microscopy, have evidence that HcB is transcytosed through enterocytes cells by an endocytosis dynamin dependant. HcB targets acetylcholinergic nerves localized in lamina propria of villi then reaches serotoninergic and acetylcholinergic nerve endings in the submucosa and musculosa within 10 minutes. In vitro experiments performed on intestinal cell line (m-ICcl2) shows that the endocytosis of HcB is dependent on the GD1b/GT1b gangliosidic receptors on the cell surface but not on the synaptotagmine II protein which is recquiered HcB entry in neuronal cells

Les toxines botuliques (BoNTs) sont les substances les plus toxiques connues. Elles sont responsables du botulisme, une maladie rare mais le plus souvent mortelle sans prise en charge médicale. Cependant, les applications médicales des BoNTs sont de plus en plus nombreuses du fait de leurs propriétés paralysantes. Leur toxicité par voie inhalée en fait un des 6 principaux agents du risque intentionnel. Les BoNTs, produites par Clostridium botulinum, se répartissent en 7 types sérologiques qui se déclinent en sous-types. Cette biodiversité rend difficile leur identification par les méthodes classiques utilisées pour les toxines protéiques (approches immunologiques). Jusqu’à présent, seule l’analyse génétique permettait de distinguer les différents sous-types entre eux. Dans ce travail a été développée une méthode d’analyse en LC-QqQ-MS/MS en mode MRM pour identifier les différents sous-types de la BoNT/A dans des matrices complexes à partir de peptides communs et spécifiques à ces sous-types. Un traitement d’échantillon par immunocapture sur billes magnétiques couplées à des anticorps anti-peptides a été développé pour isoler la toxine de l’échantillon avant analyse. Des surnageants de culture des sous-types A1 à A3, A5, A7 à A8 ont été utilisés pour valider la méthode. La limite de détection de la méthode est compatible avec les taux de toxine retrouvés habituellement dans les échantillons naturellement contaminés. Cette méthode de spectrométrie de masse a ensuite été utilisée pour quantifier les différents sous-types de la BoNT/A dans une matrice complexe (surnageants de culture de C. botulinum). Une technique de quantification, utilisant un isotope stable de la chaine légère de type A1, ([13C6]K et [13C6]R), a été retenu comme étalon interne. Les différents sous-types de BoNT/A ont été quantifiés dans les surnageants et la quantité de BoNT correspondante à une dose létale minimale de 100% a été déterminée pour chaque sous-type. / Botulinum neurotoxins (BoNTs) are the most poisonous substances known. They are responsible for human botulism, a rare but potentially fatal disease if not quickly treated. However, BoNTs were approved for the treatment of numerous medical applications due to their temporary paralysis effects. BoNTs are among the six agents with the highest risk of potential use as bio-weapons because of their high toxicity in aerosol form. BoNTs, produced by Clostridium botulinum, are divised into seven toxinotypes and each toxinotype contains several subtypes. This biodiversity makes more difficult their identification with classical methods by immunological ways. Until now, only molecular genetical methods could differenciate subtypes among them. The aim of this work was to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) in MRM mode to efficiently discrimate the distinct subtypes from specific and common peptides. Immunocapture sample preparation with antipeptides antibodies was used and allowed the isolation of the toxin from the sample. Subtyping was performed with crude supernatants (BoNT/A1 to /A3, /A5, /A7 and /A8) in order to validate the method. Limit of detection (LOD) of the proposed method is in the range of minimal toxin concentration found in naturally contamined samples. In a second part of this work, this mass spectrometry method was used to quantify the neurotoxin in complex matrices (supernatants of Clostridium botulinum cultures). Isotope labeled light chain (13C6]K et [13C6]R) from botulinum A1 neurotoxin was produced and used as internal standart. Subtypes were quantified in supernatants and the quantity of neurotoxin for one minimal lethal dose 100% was determined for each subtype

Aim: A study to investigate the potential feasibility (including estimated effect-size), acceptability and health benefits of DEFO and physiotherapy in treatment of spasticity following intramuscular injection of BT. Participants: Adults living in the community with focal spasticity of the upper or lower limb (Modified Ashworth Scale 2-3) recruited at a regional Spasticity Clinic. Intervention: provision of an individually fitted DEFO (worn daily up to 8 hours) usual care and physiotherapy (as required) for 6 weeks. Methods: Mixed methods embedded design feasibility study: Quantitative: Feasibility single-blind RCT: Intervention Group: DEFO intervention protocol, usual care and physiotherapy, Control Group: usual care and physiotherapy. Qualitative: Topic guided interviews of the intervention group and clinicians. Measures: Goal Attainment Scale (GAS) primary measure and secondary measures for function and care benefit; Arm Activity measure (ArmA), Leeds Arm Impact Score (LASIS), VAS for pain, European Quality of Life-5 Dimensions (EQ-5D), gait velocity (10MTT). Variance and fidelity was captured with: DEFO wearing record, Activity Log, clinical records and Physiotherapy modalities. Analysis: ANCOVA adjusted means and statistical comparison for significance of measures (at baseline, after six weeks and twelve weeks) between groups and to inform power calculations. Thematic Analysis of clinician and participant transcribed interviews. Quantitative and qualitative findings were integrated and triangulated to inform a larger study. Results: Participants (n=25) recruited over twelve months, (n=22) completed study. Statistical analysis showed improvements in both groups with greater health benefit in the intervention group with mean difference in the GAS of 12.17 (95% CI: 3.16 to 21.18; p = 0.014) but no statistical significance in the secondary measures. Effect-size was estimated from the GAS findings for 200 per group for a larger study. Physiotherapy modalities for spasticity were linked to 'passive' and 'active' function. Feasibility and acceptability was established with Thematic Analysis providing valuable insight into patient and clinician perspectives on disability. Conclusions: Findings indicated potential added health benefits including carer benefit. Feasibility, acceptability and clinical application of DEFO as a potential new intervention were established. This has implications for future spasticity management with patient benefit for passive and active function. Further research is indicated with a fully powered study (based on the GAS sample results) to evaluate DEFO efficacy in people with spasticity following BT.

Cosmetic treatment with botulinum toxin type A injections is the top non-surgical cosmetic procedure in the U.S. Many patients are beginning treatment at a younger age to prevent the development of facial wrinkles associated with aging. However, there is limited data to support the use of prophylactic botulinum toxin injections. Patients beginning treatment at a younger age have fewer wrinkles requiring fewer units to treat, which reduces the overall cost of treatment. Patients also maintain higher levels of self- esteem by preventing or delaying the onset of facial wrinkles that can negatively impact their appearance. This study proposes that patients receiving botulinum toxin injections at a younger age (< 35) will have higher satisfaction with treatment outcomes. By demonstrating an association between starting age of injections and patient satisfaction, this study aims to provide merit for clinical trials studying the effectiveness of prophylactic botulinum toxin injections for cosmetic indications.

Extended shelf-life cook-chill meals can pose a potential risk of botulism if they are subjected to a temperature abuse. Spores of group II non-proteolytic Clostridium botulinum can survive the mild heat treatment typically given to these products and can grow at refrigeration temperatures. To circumvent this safety issue, existing preservation methods can either affect the sensory properties of these foods or damage their image. Therefore, additional natural preservation hurdles are needed. Thus, the aim of this study was to develop a novel bio-preservation method based on the principle of antibiosis between protective cultures (PCs) and C. botulinum. Consequently, the objectives were to select effective anti-botulinal cultures and study their inhibition pattern in microbiological media and foods, identify the conditions for effective inhibition and the nature of the antibiosis. This research demonstrates for the first time that the bacteriocinogenic protective cultures inoculated at high levels had an anti-botulinal effect in a range of commercial cook-chill products, which supported active growth of non-proteolytic C. botulinum. The protocol for commercial application of the protective cultures was developed. / Doctor of Philosophy (PhD)

Thesis (MOccTher.--Faculty of Health Sciences)-University of Pretoria, 2006. / Includes bibliographical references.

The application of infrared imaging employing focal plane array Fourier transform infrared (FPA-FTIR) instrumentation for the identification of bacteria was investigated. FPA-FTIR spectroscopy was shown to provide new opportunities for bacteria identification with unprecedented reliability and throughput by allowing 102--103 FTIR spectra to be acquired simultaneously from surface areas of 90 x 90 to 200 x 200 mum with a spatial resolution of &sim;6 mum. The combination of data redundancy and spatial resolution afforded by infrared imaging made it possible to acquire highly reproducible spectra from bacterial films. A protocol for enhancing the reliability of bacteria identification by transmission-mode FPA-FTIR spectroscopy was developed by optimizing spectral acquisition parameters, spectral processing and data analysis; using the differentiation of two Campylobacter species as a test case. The results for this test case were compared with those obtained from three alternate FTIR spectral acquisition modes. The optimized protocol was employed for the generation of a spectral database of foodborne bacteria, containing over 1,000,000 spectra acquired by infrared imaging of 36 species from 19 genera. The development of a modular hierarchical clustering (MHC) model, in combination with the use of a region selection algorithm, allowed all species in the database to be differentiated from each other down to the species level based on differences in their infrared absorption profiles. A validation study involving the identification of well-characterized isolates by comparison of their spectra to those in the database demonstrated the robustness of the MHC model. In a further study employing 44 strains of Clostridium botulinum, the discriminatory power of FPA-FTIR spectroscopy was compared with that of pulsed-field gel electrophoresis, and the region selection algorithm was applied to identify growth medium-independent spectral regions that allowed for the differentiation of Group I and Group II C. botulinum strains in two blind validation studies. The research carried out also demonstrated the high-throughput potential of bacteria identification by infrared imaging when combined with the use of a microarray system for sample deposition. Overall, the novel FPA-FTIR spectroscopy-based bacteria identification protocol developed in this work provides a rapid-response and reagent-free technique suitable for routine use in both food and clinical microbiology laboratories.

Initial challenge studies with spores of proteolytic Clostridium botulinum types A and B (~104 spores/g) showed that while air- and gas-packaged English-style crumpets (aw 0.990) and pizza crust (aw 0.960) were toxic after 42-days storage at ambient temperature (25°C), no neurotoxin was detected in bagels (a w 0.944). Further challenge studies with similarly packaged crumpets inoculated with C. botulinum (~102 spores/g), pre- or post-baking, demonstrated that all crumpets were toxic within 4 to 6 days at 25°C and that toxigenesis preceded spoilage. Furthermore, reformulating crumpets to pH 8.3 and packaging in 100% CO2 had little effect in delaying the growth of C. botulinum compared to crumpets formulated to pH 6.5 and packaged in 60% CO2. / Subsequent studies were directed at determining the levels of additional barriers that could be used to ensure the safety of high-moisture MAP crumpets. While ethanol vapour proved to be an effective additional barrier in crumpets (100-g, [aw 0.990, pH 6.5]) challenged with ~102 spores/g of C. botulinum, spoilage preceded toxigenesis due to absorption of ethanol from the package headspace by crumpets. Modelling studies in Trypticase Peptone Glucose Yeast (TPGY) broth confirmed the anti-botulinal nature of ethanol and showed that a level of ~4% (vol/vol) could be used for complete inhibition of this pathogen, depending on the aw and pH of the growth medium. However, while ethanol vapour could be used to inhibit the growth of C. botulinum in high-moisture crumpets, its anti-botulinal efficacy was influenced by the method of crumpet leavening (yeast v chemical). / Preliminary studies were also done to assess the potential of mastic oil, a novel inhibitor, against C. botulinum. While direct and indirect application of ethanolic extracts of mastic oil inhibited the growth of C. botulinum in vivo, they failed to do so in crumpets.

1) Einleitung 2) Pathogenese und Klinik des Botulismus 3) Vorkommen von C. botulinum und des Botulismus 4) Beeinflussung von Spore, vegetativer Zelle und BoNT 5) Diagnostik, Therapie und Prophylaxe 6) Diskussion