Les circuits neuronaux sous-tendant la locomotion requièrent d'intégrer à la fois des stimuli sensoriels et l'état physiologique. Cependant, la manière dont ces circuits fonctionnent pendant la locomotion active reste peu comprise. La larve de poisson zèbre est un organisme vertébré idéal pour étudier cette question de part son répertoire locomoteur simple et son accessibilité à la manipulation génétique. Dans le Chapitre 1, je décris le logiciel que nous avons développé afin de nous permettre de traquer les comportements et caractériser automatiquement les modules locomoteurs à haut débit. Les interneurones V2a sont des neurones excitateurs de la moelle épinière et du cerveau postérieur caractérisés par l'expression du facteur de transcription chx10. Afin de tester leur implication dans la locomotion, j'ai, dans le Chapitre 2, validé l'utilisation d'une toxine génétiquement encodée dans le but d'inhiber la population chx10 positive in vivo. Par analyse comportementale, enregistrements de locomotion fictive et imagerie calcique, nous avons montré que les V2as sont impliqués différemment dans la locomotion lente et rapide. Les neurones contactant le liquide céphalorachidien (NcLCRs) relaient des informations sensorielles aux circuits moteurs. Par ciblage génétique, imagerie calcique, pharmacologie et électrophysiologie, j'ai, dans le Chapitre 3, investigué le rôle de l'activité spontanée dans les NcLCRs. J'ai montré que l'ouverture de canaux PKD2L1 représentait une source intrinsèque d'activité spontanée dans les NcLCRs. Ces résultats offrent une meilleure compréhension de la manière dont les interactions dynamiques structurent les sorties locomotrices in vivo. / The neural networks that underlie locomotion are complex and require integration of sensory input and physiological state. However, how these networks function during active locomotion to incorporate sensory input from the environment and the internal state of the animal remains poorly understand. The zebrafish larva is an ideal vertebrate to study these questions thanks to its simple locomotor repertoire, transparency, and amenability to genetic manipulation. In Chapter 1, I describe a program to track behavior at high speeds and automatically characterize locomotor patterns in a high-throughput manner. V2a interneurons are excitatory interneurons in the spinal cord and hindbrain identified by the chx10 transcription factor. In Chapter 2, I validated the use of a genetically-encoded botulinum toxin to silence the chx10 population in vivo. Using fictive locomotor recordings and calcium imaging, I demonstrated that silencing V2as leads to decreased activity in primary motor neurons during fast swimming, corresponding to a lower swimming frequency in V2a-silenced larvae. Cerebrospinal fluid-contacting neurons (CSF-cNs) are intraspinal neurons that relay sensory information to motor circuits. CSF-cNs in diverse species express GABA and the transient receptor potential channel PKD2L1. In Chapter 3, I used genetic targeting, calcium imaging, pharmacology, and electrophysiology to investigate the role of spontaneous activity in CSF-cNs. I showed that single channel opening of PKD2L1 represents an intrinsic source of spontaneous activity in CSF-cNs. These tools and results will allow a more complete picture of how dynamic interactions shape locomotor output in vivo.

Objective To investigate the effect a supination splint would have on upper limb function of cerebral palsy children for six months after receiving Botox® injections. Design Ten children attending weekly therapy enrolled in this prospective Quasi-experimental design where each child acted as his own control. Intervention was a supination splint and stretch massage. Assessment was based on pre- and post-intervention records of Modified Ashworth Scale, goniometry, Quality of Upper Extremity Skills Test (QUEST), and an independent panel assessment of videotaped records of hand function. Results Results show that spasticity declined in the forearm pronators, wrist flexors and thumb adductors. Active movement improved significantly in forearm supination and wrist extension. The QUEST demonstrated a significant change. Improvement in the hand function assessment was evident from the second month. Conclusion Findings support the premise that the supination splint is effective in improving upper limb function of cerebral palsy children after Botox® injections. / Dissertation (M (Occupational Therapy))--University of Pretoria, 2007. / Occupational Therapy / M (Occupational Therapy) / unrestricted

Orientador: Newton Kara-José / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-18T15:02:59Z (GMT). No. of bitstreams: 1 Minguini_Nilza_D.pdf: 5086893 bytes, checksum: 39a18efb37934b146ddf768d9cae5161 (MD5) Previous issue date: 2011 / Resumo: Os objetivos do estudo foram pesquisar a existência de efeito coadjuvante da injeção de 5U de toxina botulínica A intraoperatória em cirurgia monocular de retrocesso-ressecção para a correção de estrabismos horizontais de ângulos grandes em adultos e descrever tipos e frequências de efeitos colaterais desse tratamento. Desenho do estudo: ensaio clínico controlado randomizado duplo-cego. Vinte e três portadores de desvios horizontais de ângulos grandes foram randomizados em dois grupos: cirurgia associada à injeção intraoperatória de Toxina Botulínica A (CG+TBA) ou apenas cirurgia (CG). Os procedimentos de retrocesso-ressecção foram conduzidos sob anestesia local no olho de pior visão. A injeção de toxina botulínica A foi aplicada no músculo submetido a procedimento de retrocesso após sua tenotomia. Para a avaliação de resultados, foram comparados entre os grupos: as características clínicas, as quantidades de desvio corrigido no último retorno (6 a 12 meses) e entre o primeiro e último retornos, as porcentagens de desvios corrigidos nos retornos de 1 dia, 7 a 15 dias, 1 mês, 3 meses e 6 a 12 meses e as porcentagens de resultados satisfatórios no último retorno. As médias e desvios-padrão dos ângulos de desvio pré-operatório para os grupos CG+TBA (12 pacientes) e CG (11 pacientes) foram, em dioptrias prismáticas (Dp), respectivamente: 65,8 ± 14,9 e 60,0 ± 16,8 (p=0,26) e as médias e desvios-padrão das quantidades de desvio corrigido em Dp no último retorno foram, respectivamente, 51,5 ± 14,8 e 41,3 ± 14,0 (p=0,18). As porcentagens de desvio corrigido foram significativamente maiores no grupo CG+TBA no retorno de 30 dias (p=0,05), nos demais retornos não houve diferenças significativas entre os grupos. As quantidades de desvio corrigido foram significativamente menores no último retorno em relação ao primeiro para ambos os grupos (p=0,006) e sem diferença entre eles (p=0,59). Não houve diferença significativa entre as porcentagens de resultados satisfatórios entre os grupos no último retorno (p=0,63). Os efeitos colaterais da toxina botulínica A mais frequentemente encontrados foram: ptose e desvio vertical transitórios (em 5 dos 12 pacientes tratados). Assim, a injeção intraoperatória de TBA não aumentou o efeito cirúrgico da correção de estrabismos horizontais de ângulos grandes nos pacientes estudados e provocou alta incidência de ptose palpebral e desvio vertical transitórios / Abstract: Objective: To investigate the existence of adjuvant effect of botulinum toxin A (5u intraoperative injection) in monocular recess-resection procedure for correction of large angle horizontal strabismus in adults under local anesthesia and to describe types and frequencies of side effects. Subjects and Methods: A randomized clinical trial. Twenty-three patients with large angle horizontal deviations were randomized into two groups: surgery associated with intraoperative injection of botulinum toxin A (CG+BTA) or surgery only (CG). The recess-resection procedures were performed under local anesthesia on the non-fixating eye. The clinical characteristics, amounts of corrections at the last visit (6 to 12 months) and between the first and last visits, the percentages of changes in the pre-operative deviations at 1 day, 7 to 15 days, 1 month, 3 months and 6 to 12 months, the amount of correction between the first and last post operative visits, and the percentages of satisfactory results were compared between groups at the last visit. Results: Means and standard deviations of the preoperative angles of deviation for the groups CG+BTA and CG were 65.8 Dp ± 14.9 and 60.0 Dp ± 16.8 (p=0.26) and the means and standard deviations of the amounts of corrections at the last visit were respectively: 51,5 Dp ± 14,8 and 41,3 Dp ± 14,0 (p=0.18). The percentages of changes in the pre-operative deviations were significantly higher in the CG + TBA group at 30 days (p=0.05), and did not differ between groups at others post operative visits. Amounts of corrections were significantly lower at the last post operative visit in relation to the first for both groups (p=0.006) with no difference between them (p=0.59). There was no difference between groups in the percentage of satisfactory results at the last visit (p=0,63). Side effects of botulinum toxin A more common were: transient ptosis and transient vertical deviation both in 5 of 12 patients treated. Conclusions: Intraoperative injection of TBA did not increase the effect of surgical correction of horizontal strabismus large angles in the patients studied and led to high incidence of ptosis and vertical deviation transitory / Doutorado / Oftalmologia / Doutor em Ciências Médicas

Made available in DSpace on 2016-07-18T17:53:17Z (GMT). No. of bitstreams: 1 Gabriel Montoro Nicacio.pdf: 4088906 bytes, checksum: 508e88d0a09b6a25c4f8b77058547fe5 (MD5) Previous issue date: 2015-11-25 / This study aims to determine the efficacy of intra-articular (IA) administration of botulinum toxin type A (BoNT/A) in dogs with signs of chronic pain associated with hip dysplasia. In a double-blind design fourteen dogs were randomized and distributed into two groups: BoNT (n=7): IA injection with 25U (0,5 mL) of botulium toxin; Control (n=7): IA injection with saline solution (0.5 mL). All dogs received conventional treatment with oral and carprofen (2.2 mg/kg, every 12 h, 15 days), and chondroitin sulfate (750-1000 mg, every 12 h, 90 days). The clinical signs of HD were evaluated prior to treatment (baseline), 15, 30, 60, and 90 days after the IA injection by the veterinary using a score system and by the owners with a questionnaire about their dog s condition using the Canine Brief Pain Inventory (CBPI) and Helsinki Chronic Pain Index (HCPI). The data were analyzed using test unpaired-t, ANOVA, Tukey test (P < 0.05). There was no difference between groups in the scores measured by the veterinary and by the owners (CBPI and HCPI). In comparison over time lower scores were observed in both groups during 90 days from baseline in the researcher evaluation and in the HCPI. The same result was obtained by the CBPI evaluation to the control group whereas for the BoNT group the difference was only observed in the first 60 days after IA injection. Analgesic intervention was not necessary during the evaluation period. Both treatments reduced the clinical signs associated with hip dysplasia, however adjunctive administration of BoNT didn t potentialize the results. / Objetivou-se avaliar a administração intra-articular (IA) da toxina botulínica tipo A (BoNT/A) como adjuvante do controle da dor crônica em cães com displasia coxofemoral (DCF). Em delineamento duplo-cego, 14 cães foram distribuídos aleatoriamente em dois grupos: BoNT (n=7): administração IA de 25U (0,5 mL) de toxina botulínica; Controle (n=7): administração IA de 0,5 mL de solução salina. Para todos os animais foi prescrito tratamento convencional com carprofeno (15 dias) e sulfato de condroitina (90 dias). Os sinais clínicos da DCF foram avaliados, antes do tratamento (basal), 15, 30, 60 e 90 dias após a injeção IA, por sistema de escore pelo pesquisador e mediante questionários respondidos pelos proprietários dos cães, empregando-se o Breve Inventário de Dor Canina (BIDC) e o Indicador de Dor Crônica de Helsinque (IDCH). Intervenção analgésica foi permitida se o somatório dos escores do BIDC e/ou IDCH excedesse 50%. Os resultados foram analisados pelo teste t não-pareado, ANOVA, teste de Tukey (P < 0,05). Não houve diferença entre os grupos nos escores avaliados pelo médico veterinário ou pelos proprietários (BIDC e IDCH). Na comparação ao longo tempo, escores inferiores foram observados em ambos os grupos durante 90 dias em relação ao basal na avaliação do pesquisador e no IDCH. O mesmo resultado foi obtido na avaliação pelo BIDC para o grupo controle, enquanto no grupo BoNT a diferença só foi observada nos primeiros 60 dias após a injeção IA. Intervenção analgésica não foi necessária durante o período de avaliação. Ambos os tratamentos reduziram os sinais clínicos associados à DCF, porém a administração adjuvante de toxina botulínica não potencializou os resultados.

Submitted by Nadir Basilio (nadirsb@uninove.br) on 2018-06-19T15:38:25Z No. of bitstreams: 1 Maria Eliege de Souza.pdf: 1276931 bytes, checksum: dc49846f1db7db8cc9034d54ff988caf (MD5) / Made available in DSpace on 2018-06-19T15:38:25Z (GMT). No. of bitstreams: 1 Maria Eliege de Souza.pdf: 1276931 bytes, checksum: dc49846f1db7db8cc9034d54ff988caf (MD5) Previous issue date: 2015-12-14 / The cerebral palsy (CP) is characterized by a group of non progressive disorders of the brain’s development and posture caused by a malformation or brain injury. As treatment , there are therapeutic approaches aimed at normalizing muscle tone and prevent change in the relation between bone growth and muscle, that can lead to poor posture and structured deformities. The aim this study was to investigate the effects of botulinum toxin type A (TB-A) for spasticity and functionality in spastic CP childrem. This was a prospective, controlled, randomized, consisting of 24 children (aged 5 to 12 years) with spastic CP, being divided into two groups: experimental group (EG) consisting of 12 patients (mean age 7.83 ± 2, 07 years) treated with TB-A toxin and physiotherapy and control group (CG) with 12 patients (mean age 8.50 ± 2.17 years) treated only with physical therapy. All participants were assessed through motor and functional scales (GMFCS, GMFM-88, Ashworth, Berg Balance Scale, Time up and go -TUG and Inventory Assessment Pediatric Disorders - ASK) at three different times: before treatment, 30 days and 3 months after the treatment proposed. By analysis of variance (ANOVA) for repeated measures it was observed significant differences (p < 0.001) between groups (groups vs treatment) to the data obtained in the GMFCS, GMFM-88, BERG, TUG, Ashworth and ASK, being observed functional improvement only for the GE group. In this study it was concluded that the use of TB-A provides a significant improvement on spasticity and child functionality with spastic CP, over a period of up to 3 months after application. / A Paralisia Cerebral (PC) é caracterizada por um grupo não progressivo de desordens do desenvolvimento e postura decorrentes de uma malformação ou lesão cerebral. Como tratamento, destacam-se abordagens terapêuticas que visam normalizar o tônus muscular e prevenir alteração na relação entre o crescimento ósseo e o muscular, que podem provocar posturas inadequadas e deformidades estruturadas. Dessa maneira, o objetivo desse estudo foi verificar os efeitos da toxina botulínica tipo A (TB-A) sobre a espasticidade e funcionalidade da criança com PC espástica. Esse foi um estudo prospectivo, controlado, randomizado, constituído por 24 crianças (idade entre 5 e 12 anos) com PC espástica, distribuídas em dois grupos sendo: Grupo experimental (GE) constituído com 12 pacientes (idade média 7,83  2,07 anos) tratadas com toxina TB-A e fisioterapia e Grupo controle (GC) sendo 12 pacientes (idade média 8,50  2,17 anos) tratados somente com fisioterapia. Todos os participantes foram avaliados por meio de escalas motoras e funcionais (GMFCS, GMFM-88, Ashworth, escala de equilíbrio de BERG, Time up and go –TUG e o Inventário de Avaliação Pediátrica de Disfunções - PEDI) em três momentos distintos: antes do tratamento, 30 dias e 3 meses após o tratamento proposto. Por meio da análise de variância (ANOVA) para medidas repetidas foi possível observar diferenças significativas (p < 0,001) entre os grupos (grupos vs tratamento) para os dados obtidos na escala de GMFCS, GMFM-88, BERG, TUG, Ashworth e PEDI, sendo observada melhora funcional somente para o grupo GE. Nesse estudo foi possível concluir que o uso da TB-A proporciona uma melhora significativa sobre a espasticidade e funcionalidade da criança com PC espástica, em um período de até 3 meses após sua aplicação.

In the 1950s, the discovery of anti-nerve growth factor, an immunotoxin stunting sympathetic neural development, signaled the advent of neurotoxins as research modalities. Other selective neurotoxins were discovered in rapid succession. In the 1960s, 6-hydroxydopamine and 6-hydroxydopa were shown to destroy noradrenergic and dopaminergic nerves. Excitotoxins (glutamate, aspartate, and analogs) were discovered in the 1970s. DSP-4 [N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine] proved to be selective for noradrenergic destruction, while 5,6-and 5, 7-dihydroxytryptamines were relatively selective for serotonin neurons. Additional neurotoxins were discovered, but it was MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) that predominated neurotoxicity research in the 1980s. Eventually, Clostridium botulinum neurotoxin (BoNT), discovered as a "poisonous" principle in the late 1800s, resurfaced in purified and standardized forms as a clinically useful drug. Neurotoxins represent chemical tools, useful not only for discerning neuronal mechanisms and animal modeling of neurological disorders, but also for their use in medicine and potential as treatments for medical disorders. This unit reviews the early discovery of neurotoxins, describes categories of neurotoxins, and finally characterizes their usefulness - first as research tools, and eventually as clinical therapeutic agents.

Botulinum neurotoxin (BoNT), for more than a hundred years, has been a recognized poisonous principle in spoiled food. As its chemical structure became unraveled, and as more knowledge was gained over its mechanism of toxicity, it became clear that BoNT had the potential to act therapeutically as a targeted toxin that could inactivate specific nerve populations, and thus achieve a therapeutic goal. BoNT has evolved over the past 25 years into a viable therapeutic, now being a first line treatment for dystonia, overtly altering the course of progression of this disorder. BoNT is used for hyperhidrosis and gustatory sweating syndrome, alleviation of pain, as a treatment for overactive bladder, achalasia and anal fissure; and it has gained popularity as a cosmetic aid. Many other possible uses are being explored. The greatest potential for BoNT may lie in its being a molecular Trojan Horse - able to carry a specific enzyme or specific drug to the inside of a cancer or other type of cell while bypassing other cells and thereby having little or no ill effect. BoNTs pharmaceutical potential is boundless.

The growth of Listeria monocytogenes and nonproteolytic Clostridium botulinum type B spores in cooked, uncured turkey was investigated separately under varying conditions of modified atmosphere packaging (MAP), refrigerated and temperature-abuse storage, and lactic acid bacteria (LAB) competition. L. monocytogenes (LM) growth was suppressed when initially outnumbered 3.5 logs:1 or 2.1 logs:1 by naturally-occurring LAB, but not when the initial LAB:LM population ratio was equivalent. Lowering storage temperature from 10 degrees to 4 degrees C enhanced the inhibitory effect of CO₂ in the packaging atmosphere, and extended the period of product olfactory acceptability. When the LAB:LM population ratio was equivalent, objectionable odors were not detected in most of the samples, despite LAB counts in excess of 10E⁸/g. This raises concerns with respect to public health, since high levels of L. monocytogenes can be present in MAP meat and poultry products without accompanying signs of overt spoilage. Cellular fatty acid (CFA) analysis was a valuable tool for distinguishing between phenotypically distinct isolates of LM inoculated into MAP turkey. Fatty acid composition of foodborne outbreak-associated (serotype 4) and non-outbreak-associated (serotype 1) strains of LM correlated with antigenic type (4 or 1) and agglutination reaction (granular or flocculent). Strain ATCC 43256 (serotype 4b) produced a consistently unique CFA profile, making it the easiest of the four test strains to be differentiated. Analysis of additional LM serotypes, as well as examination of existing clinical and environmental CFA databases for correlations between fatty acid profiles and diagnostic characteristics of LM, is necessary before CFA analysis can be effectively applied as an epidemiological tool for tracking the distribution of LM strains in food products and throughout the farm-to-table food chain. Reduced storage temperature significantly delayed botulinal toxin production and extended the period of olfactory acceptability of cooked turkey, but even strict refrigeration did not prevent growth and toxigenesis by nonproteolytic C. botulinum type B. Toxin was detected on day 7 for product stored at 15 degrees C and on day 14 for product stored at 10 degrees C, irrespective of packaging atmosphere. At 4 degrees C, toxin was detected on day 14 for product packaged without carbon dioxide, and on day 28 for product packaged with 30% carbon dioxide. At all three storage temperatures, toxin detection preceded or coincided with olfactory unacceptability, demonstrating the potential for consumption of toxic product when spoilage-signalling sensory cues are absent. / Ph. D.

Indiana University-Purdue University Indianapolis (IUPUI) / Neurofibromatosis Type 1 (NF1) is a highly penetrant autosomal dominant genetic disorder where mutations in the tumor suppressor gene NF1 leads to decreased neurofibromin. The most debilitating manifestation is the presence of complex multilineage Schwann cell-derived plexiform neurofibromas (PN). Historically, little clinical success has been achieved targeting PN through surgery or chemotherapies. I performed an shRNA library screen of patient-derived Schwann cell lines to identify novel therapeutic targets to disrupt PN formation and progression. An shRNA library screen of human kinases and Rho-GTPases was performed in NF1-/- and paired NF1 competent immortalized Schwann cell lines. Following sequencing, candidates were identified. We previously developed a novel mouse model of NF1 wherein a neural crest specific Postncre targeted loxp-flanked Nf1 that replicated the PN found in patients. Additional cohorts of mice were generated with biallelic deletion of Rac1 (Nf1f/fRac1f/f Postn-Cre+; DKO ). Mice were aged for 9 months and peripheral nerves were harvested and fixed in formalin. Peripheral nerve size was measured and tumors were identified through blinded analysis of hematoxylin and eosin and Masson’s Trichrome (collagen) stained slides. Rho family members, including RAC1, were identified as candidates through an shRNA library screen. Genetic disruption of Rac1 in the Schwann cell lineage resulted in the prevention of tumor formation in DKO mice, as observed by peripheral nerve size and histological analysis. I observed an average of 14.8 +/- 2.65 tumors per mouse in the Nf1f/f Postnviii Cre+ cohort compared to 0 tumors in the DKO (p<0.0001). Following an shRNA library screen, RAC1 was identified as a candidate to modulate PN formation. Biallelic deletion of Rac1 in vivo prevented PN formation. I demonstrate that a candidate identified in an shRNA library screen can translate to an biological effect in a mouse model of PN.

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