Formål: At undersøge, hvordan forældre til børn med spastisk cerebral pare­se (CP) oplevede effekt af behandling med Botulinumtoxin type A (BTX) på deres barns funktioner. Metode og materiale: Der blev anvendt triangulering af metode, kilder og observatører. Den kvali­ta­tive del bestod af to fokusgruppeinterview af eksperter og tre forælderinterview. Analyseme­toden var indholdsanalyse. Den kvantitative del bestod af en spørgeskemaundersøgelse til forældre til børn behandlet med BTX på Hillerød Hospital. Spørgeskemaer blev sendt til begge forældre til 47 børn. Resultat: Fokusgruppeinterviewene gav indblik i, at der i spørgeskemaet skulle spørges detaljeret til alle funktioner i barnets hverdag fra BTX blev tilbudt til efter endt effekt af BTX-behandling. Forældreinterviewene viste, at to af tre forældre ville tage imod et nyt tilbud om BTX-behandling; at ventetid på BTX-dagen var uacceptabel lang; at arbejdsgange på BTX-dagen burde optimeres, så de passede til handicappede børn. Spørgeskemaet blev returneret af 41 forældre til ialt 30 børn (to – 17 år). Børnene var behandlet med BTX på Hillerød Hospital i perioden 2004-2007. Af de 41 forældre var der 12 fædre og 29 mødre. Størst effekt af BTX oplevede forældrene på nedsættelse af barnets spasticitet. To tredjedele af de børn, som kunne gå før BTX behandling, fik forbedret gang. Derud­over sås effekt i nye måder at bevæge sig på. Toogtyve af 41 forældre så den effekt af BTX, som de havde fået stillet i udsigt. Størst effekt af BTX sås indenfor tre til seks måneder. Virkningen af BTX for det enkelte barn var forskellig fra behandling til behandling med en dalende tendens. Studiet viste, at forældre og børn blev godt mødt og blev hørt på børneafdelin­gen, var trygge ved persona­let, og oplevede BTX-dagen som god. Ventetid og et manglende måltid mad efter BTX var et stort pro­blem. Der sås ingen forskel i fædres og mødres opfattelse af sundhedsydel­ser til børn med CP. Nationale retningslinier for behandling med BTX ef­terspørges. Konklusion: Majoriteten af forældre ville tage mod et nyt tilbud om behandling med BTX. De væ­sentligste begrundelser var, at forældrene havde set god effekt af behandlingen, eller at de håbede på en effekt. Et mindretal af forældre ville ikke tage imod et nyt tilbud om behandling med BTX, fordi de enten ikke havde set effekt af behandlingen, eller fordi de oplevede mange strabadser for et meget lille udbytte. Praksis omkring BTX dagen bør udvikles og forbedres i forhold til handicappede børn / This study aimed to assess how parents of children with spastic Cerebral Palsy (CP) evaluated the efficacy of Botulinum Toxin A (BTX) treatment on their child. Method: Triangulation of measurement processes, i.e., qualitative assessment including two focus group interviews with experts and three interviews with parents; quantitative assessment involving questionnaires send to parents of 47 children treated with BTX at Hilleroed Hospital. The qualitative data were analysed using content analysis. Results: Focus group interviews indicated that questionnaires should seek detailed information about the daily activities of children undergoing BTX treatment, beginning treatment initiation and continuing until BTX effect declined. Parent interviews showed that two out of three would allow another BTX injection; waiting time on treatment day was too long; and hospital routines should be optimized to meet the needs of disabled children. Forty-one parents (12 fathers and 29 mothers) completed the questionnaire. The study included 30 children receiving BTX between 2004 and 2007. Parents described the best effect of BTX as reduced spasticity. Walking ability improved in 67% of children who could walk before treatment, and improved motor skills provided new ways of moving. Twenty-two parents believed that BTX treatment fulfilled doctors’ expectations. Maximum BTX effect occurred after three to six months. Effects differed with each treatment, mostly on a declining scale. Parents felt very welcome in the children’s ward and also felt that the staff listened to them; their overall assessment of treatment day experience was good. Major concerns included waiting time and a missing meal following treatment. Views of healthcare services for children with CP did not differ between fathers and mothers. The study indicates that development of national guidelines for BTX treatment will be highly recommended. Conclusion: The majority of parents agreed to repeat BTX injections, mainly because outcome was positive or met their expectations. A minority would refuse future BTX treatment, primarily due to lack of effect or because of significant problems for small benefit. Optimizing hospital routines on treatment day would help disabled children navigate BTX treatment more successfully / <p>ISBN 978-91-85721-52-8</p>

Orientador: Wilter Ricardo Russiano Vicente / Banca: Maria Denise Lopes / Banca: Maria Isabel de Mello Martins / Resumo: A hiperplasia prostática benigna (HPB) tem início no animal com um a dois anos de idade, sendo que 80% dos cães com cinco anos apresentam evidências histológicas de sua presença. A fisiopatologia da doença não está totalmente compreendida, no entanto, a diidrotestosterona é o principal hormônio envolvido. Recentemente, o efeito da toxina botulínica A (TB-A) foi investigado na próstata, mostrando que esta induz atrofia do parênquima e redução do volume. Como o cão é o único animal doméstico que apresenta esta alteração, este se apresenta como modelo experimental para novos estudos da HPB humana. Com base nisso, este estudo objetivou fornecer informações acerca dos efeitos da TB-A sobre a próstata, libido e qualidade do sêmen, comparando os dados com animais orquiectomizados. Para tanto, 18 cães adultos, com evidências ultra-sonográficas de HPB foram submetidos à castração ou administração de 250 ou 500 U de TB-A, e avaliados durante 16 semanas. A orquiectomia mostrou-se um excelente tratamento para a HPB, promovendo redução de 80% do volume prostático. Aplicação da TB-A não ocasionou alterações significativas na libido, ereção ou qualidade e características seminais. Efeitos locais e sistêmicos também não foram observados. Administração de 250 U da TB-A promoveu redução máxima de 9,4% do volume prostático, entretanto, tal redução não foi significativa. Por outro lado, a administração de 500 U de TB-A reduziu significamente as variáveis comprimento, altura e volume da próstata. Desta forma, o presente ensaio contribui de forma singular e inovadora para o conhecimento dos efeitos desta nova modalidade de tratamento na HPB canina. / Abstract: Benign prostatic hyperplasia (BPH) starts the development in animals aging about 1 - 2 years. 80% of 5 years-old dogs have histologic evidences of BPH. Despite the little knowledge concerning about this disease, dihydrotestosterone is the main involved hormone. Recently, the effect of botulinum toxin A (BT-A) on rat and human prostate was investigated, and prostatic parenchyma atrophy and decrease in glandular volume were observed. The dog is one of a few animals that can develop BPH spontaneously and is frequently used as an animal model for human prostatic hyperplasia. Based on that, this study aimed to provide information on BT-A effects on prostate, libido and semen quality, in comparison to orchiectomized dogs. For that, 18 adults dogs, with Ultrasonographic evidences of BPH were submitted to orchiectomy or administration of 250 or 500 U of BT-A, and evaluated along 16 weeks. Orchiectomy presented excellent results on BPH, reducing the prostate volume up to 80%. Administration of BT-A did not significantly interfered on libido, erection or semen characteristics. Local and systemic effects also were not observed. Administration of 250 U of BT-A has promoved a maximum decrease of 9,4% on prostatic volume. However, this reduction was not statistically significant. On the other hand, 500 U of BTA administration has shown to significantly reduce the length, height and volume of prostate. This way, the present study is an innovative and singular contribution for the knowledge of the effects of BT-A on canine prostate. / Mestre

INTRODUÇÃO: O objetivo do presente estudo é apresentar dados morfométricos da fenda palpebral e da função do músculo levantador da pálpebra superior, ao longo de 6 meses de seguimento, após a aplicação da toxina botulínica tipo A, em pacientes com retração palpebral nas fases aguda e crônica da orbitopatia distireoidiana. MÉTODOS: Estudo prospectivo, no qual foram incluídos 24 indivíduos, divididos em dois grupos, agudos (n=12) e crônicos (n=12). Foram submetidos à aplicação de toxina botulínica na dose de 5 UI/0,1 ml em uma das pálpebras superiores e seguidos por 6 meses. A avaliação palpebral foi registrada por meio de captação de imagens com uma câmera filmadora de vídeo conectada a um microcomputador. Os valores médios obtidos nos diferentes momentos analisados foram comparados entre os dois grupos. RESULTADOS: A maioria dos pacientes apresentou melhora da retração da pálpebra superior, com redução média de 3,05 mm no grupo agudo e 3,81 mm no grupo crônico. Esta redução foi significativa até 1 mês no grupo agudo e até 3 meses no grupo crônico. Houve diminuição da função do músculo levantador e da diferença entre as áreas lateral e medial da fenda palpebral nos olhos tratados em ambos os grupos. No olho contralateral, houve aumento significativo da fenda palpebral até 2 semanas após a aplicação. Não houve diferença na freqüência dos efeitos colaterais entre os 2 grupos. CONCLUSÕES: A injeção de 5 unidades foi eficaz na redução da retração e da função do músculo levantador e na melhora do contorno palpebral superior, nos pacientes na fase aguda e crônica da orbitopatia distireoidiana, embora esta melhora tenha sido menos duradoura no grupo agudo / INTRODUCTION: The objective of this study is to compare morphometric data of the eyelid fissure and the levator muscle function after transcutaneous injection of botulinum toxin type A with six-month follow-up in patients with upper eyelid retraction from acute and chronic dysthyroid orbitopathy. METHODS: This was a prospective study that included 24 individuals with dysthyroid orbitopathy, divided into two groups, acute (n=12) and chronic (n=12). They were given a 5 UI/0,1 ml dose of botulinum toxin in the upper eyelid and examined periodically for 6 months. Clinical eyelid examination was performed and recorded using a video camera connected to a microcomputer. Mean values taken at different follow-up points were compared for the two groups. RESULTS: Most patients experienced marked improvement in upper eyelid retraction, with a mean reduction of 3.05 mm in acute group and 3.81 mm in chronic group. This reduction was significantly smaller during 1 month in acute group and during 3 months in chronic group. Reduction in levator function and in the difference between lateral and medial lid fissure measurements was observed in both groups. Measurements of the contralateral eye increased up to 2 weeks after the application. No difference was observed between the study groups with regard to the observed side effects. CONCLUSIONS: A single 5-unit botulinum toxin injection improved upper eyelid retraction, reduced levator function, and produced an adequate lid contour in patients with acute and chronic dysthyroid orbitopathy. The effect lasts longer in patients with chronic disease than in patients with acute disease.

Submitted by (lucia.rodrigues@ufrpe.br) on 2016-06-08T13:37:15Z No. of bitstreams: 1 Jaciel Benedito de Oliveira.pdf: 2001810 bytes, checksum: 849f2538ec8321a9c6a102564bb281e9 (MD5) / Made available in DSpace on 2016-06-08T13:37:15Z (GMT). No. of bitstreams: 1 Jaciel Benedito de Oliveira.pdf: 2001810 bytes, checksum: 849f2538ec8321a9c6a102564bb281e9 (MD5) Previous issue date: 2014-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The aim of this study was to observe the morphometric, histological and histométricos aspects of submandibular glands of rats treated with botulinum toxin type A (BTX-A) and with bovine gelatin. We used 80 submandibular glands of adult Wistar rats and divided into three groups: group I, control (n = 14); group II (n = 34) submitted the intraglandular injection of bovine gelatin (0.25 mg); and group III (n = 32) treated with intraglandular injection of botulinum toxin type A (2.5 U). The groups II and III were divided into 3 subgroups according to the day of analysis of glands (7, 15 and 28 days after treatment), when they were removed, heavy and measures immediately for morphometric analysis. Next, were fixed in paraformaldehyde 4 % and processed conventionally for light microscopy, stained in H.E., P.A.S., alcian blue, Gomori trichrome and picrosirius red. Morphometric differences were not observed between subgroups of group III, however in group II there was a significant difference between the three subgroups in the longitudinal length (p = 0.030) and transverse length (p = 0.025), having the gland has shown greater at day 28th after treatment. Histometrically, serous and mucous acini group III, were presented with a smaller diameter compared with the acinithe subgroups treated with bovine gelatina (group II). The wall thickness of the striated duct Group III was lower than in other. Histologically, there was a greater amount of cytoplasmic vacuolization in group III. addition to the greater spacing between the acini. Thus, we conclude that botulinum toxin type A interferes with the structural organization of the submandibular glands, promoting a reduction in the diameter of serous acini and in wall thickness striated ducts, macroscopically but this difference was not significant. / O objetivo deste trabalho foi observar os aspectos morfométricos, histométricos e histológicos das glândulas submandibulares de ratas tratadas com toxina botulínica tipo A (BTX-A) e com gelatina bovina. Foram utilizadas 80 glândulas submandibulares de ratas Wistar adultas e divididas em três grupos: grupo I controle (n = 14); grupo II (n = 34) submetidas a injeção intraglandular de gelatina bovina (0,25 mg); e grupo III (n = 32) tratadas com injeção intraglandular de toxina botulínica tipo A (2,5 U). Os grupos II e III foram divididos em 3 subgrupos de acordo com o dia de análise das glândulas (7, 15 e 28 dias após o tratamento), quando foram removidas, imediatamente pesadas e medidas para análise morfométrica. A seguir, foram fixadas em paraformaldeído 4% e processadas convencionalmente para microscopia de luz, coradas em H.E, P.A.S., alcian blue, tricrômico de Gomori e picrossirius red. Testes estatísticos dos resultados morfométricos e histométricos obtiveram o nível de significância de 0,05 (5%). Morfometricamente não foram observadas diferenças entre os subgrupos do grupo III, entretanto no grupo II houve diferença significativa entre os três subgrupos no comprimento longitudinal (p = 0,030) e comprimento transversal (p = 0,025), tendo a glândula se apresentado maior no 28º dia após o tratamento. Histometricamente, os ácinos serosos e mucosos do grupo III, apresentaram-se com diâmetro menor quando comparado com o ácinos dos subgrupos tratados com gelatina bovina (grupo II). A espessura da parede do ducto estriado do grupo III foi menor que nos demais. Histologicamente, observou-se a maior quantidade de vacuolizações citoplasmáticas no grupo III, além do maior espaçamento entre os ácinos. Dessa forma, concluímos que a toxina botulínica tipo A interfere na organização estrutural das glândulas submandibulares, promovendo uma diminuição no diâmetro dos ácinos serosos e na espessura da parede dos ductos estriados, mas que macroscopicamente tal diferença não se mostrou significante.

INTRODUÇÃO: A paralisia facial é constrangedora tanto do ponto de vista funcional quanto estético. Esta pesquisa teve por objetivo verificar o efeito da terapia miofuncional em pacientes com paralisia facial de longa duração associada à aplicação de toxina botulínica. MÉTODOS: Foram tratados vinte e cinco pacientes, divididos em dois grupos. Os pacientes do grupo A receberam quatro sessões de terapia miofuncional antes da aplicação de toxina botulínica e os pacientes do grupo B simultaneamente à aplicação. A terapia foi composta por manobras isométricas e isotônicas passivas, intra e extraorais, além de exercícios de resistência. RESULTADOS: Após a terapia miofuncional os pacientes apresentaram aumento significativo da mobilidade do lado paralisado da face, do índice de satisfação do paciente com a face, do Índice Funcional da Face (IFF) e do Índice de Bem-Estar Social (IBES). O grupo de pacientes que realizaram a terapia miofuncional previamente, apresentou freqüência significativamente maior de dificuldade para falar, enquanto o grupo que realizou a terapia miofuncional a partir da data da aplicação de toxina botulínica, apresentou freqüência significativamente maior de dificuldade para mastigar. A terapia miofuncional promove simetria facial; satisfação dos pacientes com a face, funcionalidade oromiofacial, qualidade de vida e deve ser realizada antes e após aplicação de toxina botulínica para reduzir os possíveis efeitos adversos. / INTRODUCTION: The facial paralysis is constraining so much of the functional point of view as aesthetic. This research had for goal verified the myofunctional therapy effect in patients with long-standing facial paralysis associate to the botulinum toxin application. METHODS: Twenty-five patients were treated, divided into two groups. The patients from the group A received four sessions of myofunctional therapy before the toxin botulinum application and the patients from the group B received it simultaneously to the application. The therapy was composed by isometric and passive isotonic maneuvers, inside and outside oral, and resistance exercises. RESULTS: After the myofuncional therapy the patients presented significant increase of the mobility of the paralyzed side, of the patient satisfaction index with the face, Functional Index of the Face (IFF) and of the Index of Social Welfare (IBES). The group of patient that accomplished the myofuncional therapy previously presented significantly larger frequency of talking difficulty, while the group that accomplished the miofuncional therapy from the toxin botulínica application date presented significantly larger frequency of chewing difficulty. The myofuncional therapy promotes facial symmetry; patients satisfaction with the face, myofuncional functionality, life quality, and should be accomplished before and after toxin botulínica application to reduce the possible adverse effects.

This Master's thesis focuses on Occupational therapy for patients after application of botulinum toxin to upper limb. The aim of the thesis was to evaluate and compare a progress of functional independence in patients after acquired brain injury. There were twenty patients involved in the research, who were further divided into two groups and separated by ten people in each group. Both groups underwent the application of botulinum toxin whereas the experimental group A went through the occupational therapy. The control group had ordinary rehabilitation at home and without any occupational intervention for the duration of twelve week under guided self-rehabilitation contract of spastic paresis, so-called GSC. A programme of the group A included over the counter rehabilitation outpatient occupational therapy. The patients attended the course twice a week for the duration of four weeks. Afterwards, they received a home programme for the duration of eight weeks, during which they had two examinations at occupational therapy. All the participants were evaluated by Global Subjective Self-Assessment which is focused on spasticity. There were further evaluated according to Functional Independece Measure and modified Frenchay Arm Test. The experimental group with outpatient occupational therapy was...

Made available in DSpace on 2016-01-26T18:55:37Z (GMT). No. of bitstreams: 1 Sergio Vilhegas.pdf: 700648 bytes, checksum: 61199638be80e4484e7e0b1e44be0387 (MD5) Previous issue date: 2013-09-23 / The aim of this study was to evaluate the analgesic effects of botulinum toxin as an adjunct in the control of postoperative pain in dogs. In a blinded study, sixteen dogs undergoing mastectomy were randomly assigned to two groups of 08 animals each and received 7U kg-1 of botulinum toxin diluted in 10mL of saline and administered 1mL in each breast, subcutaneously (TXB) or 1mL of saline administered in each breast (Control), subcutaneously. After twenty four hours, pre-anesthetic medication was intramuscular (IM) acepromazine (0.03mg kg-1) in combination with morphine (0.3mg kg-1). Anesthesia was induced with intravenously (IV) propofol (4 to 5mg kg-1) and maintained with isoflurane. The analgesic intra-operative support was provided by IV continuous rate morphine (0.1mg kg-1h-1). Heart rate, respiratory rate, systolic arterial blood pressure, oxicapnografy and end-tidal concentration of isoflurane were evaluated during the surgery. Postoperative analgesia was assessed during the first 72 hours after the tracheal extubation using a Visual Analog Scale (VAS) and Glasgow Modified Scale (GMS). Rescue analgesia with morphine (0.5mg kg-1 IM) was performed if the evaluation score exceeded 50% of VAS and/or 30% of GMS during the postoperative period. The pain scores were lower in TXB group than in Control group from 8 to 60 hours and from 12 to 60 hours post-extubation, according to VAS and GMS scales, respectively. Postoperative rescue analgesia was lower in the TXB group (2 of 8 dogs), when compared with Control group (7 of 8 dogs). Cardiopulmonary established was detected in both treatment groups. Preemptive botulinum toxin administration is a satisfactory alternative to post-operative management in dogs undergoing mastectomy. / Este estudo teve como objetivo avaliar a administração da toxina botulínica como adjuvante do controle da dor pós-operatória de cães. Em delineamento cego, dezesseis cadelas, encaminhadas para mastectomia radical foram distribuídas aleatoriamente em dois tratamentos: TXB (n=8) administração de 7U kg-1 de toxina botulínica, diluída em 10 mL de solução salina, sendo administrado 1mL em cada mama, por via subcutânea (SC); Controle (n=8) administração de 10 mL de solução salina (1mL em cada mama, SC). Vinte e quatro horas após, os animais foram encaminhados para cirurgia, sendo tranquilizados com acepromazina (0,03mg kg-1) associada à morfina (0,3mg kg-1), por via intramuscular (IM). Transcorridos vinte minutos, iniciou-se a infusão contínua intravenosa (IV) de morfina (0,1mg kg-1 h-1) que foi mantida durante toda a cirurgia. A indução anestésica foi realizada com propofol (4-5mg kg-1, IV), seguindo-se a manutenção da anestesia com isofluorano. Parâmetros avaliados: frequência cardíaca e respiratória, pressão arterial sistólica, oxicapnografia e concentração final expirada de isofluorano. No pós-operatório o grau de analgesia foi mensurado durante 72 horas após a extubação traqueal, utilizando-se a Escala Analógica Visual (EAV) e Escala de Glasgow Modificada (EGM). Analgesia de resgate foi feita com morfina (0,5mg kg-1 IM) em casos do escore de dor ser superior a 50% da EAV e/ou 33% da EGM. Os escores de dor foram inferiores no tratamento TXB de 8 a 60 horas na EAV e de 12 a 60 horas na EGM. A necessidade de suplementação analgésica foi inferior no gruo TXB (2 de 8 animais) em relação ao grupo Controle (7 de 8 animais). Estabilidade cardiorrespiratória foi observada em ambos os tratamentos. A administração preemptiva de toxina botulínica representa uma alternativa viável para o controle da dor pós-mastectomia em cadelas.

Made available in DSpace on 2016-07-18T17:53:12Z (GMT). No. of bitstreams: 1 Sergio Vilhegas.pdf: 700648 bytes, checksum: 61199638be80e4484e7e0b1e44be0387 (MD5) Previous issue date: 2013-09-23 / The aim of this study was to evaluate the analgesic effects of botulinum toxin as an adjunct in the control of postoperative pain in dogs. In a blinded study, sixteen dogs undergoing mastectomy were randomly assigned to two groups of 08 animals each and received 7U kg-1 of botulinum toxin diluted in 10mL of saline and administered 1mL in each breast, subcutaneously (TXB) or 1mL of saline administered in each breast (Control), subcutaneously. After twenty four hours, pre-anesthetic medication was intramuscular (IM) acepromazine (0.03mg kg-1) in combination with morphine (0.3mg kg-1). Anesthesia was induced with intravenously (IV) propofol (4 to 5mg kg-1) and maintained with isoflurane. The analgesic intra-operative support was provided by IV continuous rate morphine (0.1mg kg-1h-1). Heart rate, respiratory rate, systolic arterial blood pressure, oxicapnografy and end-tidal concentration of isoflurane were evaluated during the surgery. Postoperative analgesia was assessed during the first 72 hours after the tracheal extubation using a Visual Analog Scale (VAS) and Glasgow Modified Scale (GMS). Rescue analgesia with morphine (0.5mg kg-1 IM) was performed if the evaluation score exceeded 50% of VAS and/or 30% of GMS during the postoperative period. The pain scores were lower in TXB group than in Control group from 8 to 60 hours and from 12 to 60 hours post-extubation, according to VAS and GMS scales, respectively. Postoperative rescue analgesia was lower in the TXB group (2 of 8 dogs), when compared with Control group (7 of 8 dogs). Cardiopulmonary established was detected in both treatment groups. Preemptive botulinum toxin administration is a satisfactory alternative to post-operative management in dogs undergoing mastectomy. / Este estudo teve como objetivo avaliar a administração da toxina botulínica como adjuvante do controle da dor pós-operatória de cães. Em delineamento cego, dezesseis cadelas, encaminhadas para mastectomia radical foram distribuídas aleatoriamente em dois tratamentos: TXB (n=8) administração de 7U kg-1 de toxina botulínica, diluída em 10 mL de solução salina, sendo administrado 1mL em cada mama, por via subcutânea (SC); Controle (n=8) administração de 10 mL de solução salina (1mL em cada mama, SC). Vinte e quatro horas após, os animais foram encaminhados para cirurgia, sendo tranquilizados com acepromazina (0,03mg kg-1) associada à morfina (0,3mg kg-1), por via intramuscular (IM). Transcorridos vinte minutos, iniciou-se a infusão contínua intravenosa (IV) de morfina (0,1mg kg-1 h-1) que foi mantida durante toda a cirurgia. A indução anestésica foi realizada com propofol (4-5mg kg-1, IV), seguindo-se a manutenção da anestesia com isofluorano. Parâmetros avaliados: frequência cardíaca e respiratória, pressão arterial sistólica, oxicapnografia e concentração final expirada de isofluorano. No pós-operatório o grau de analgesia foi mensurado durante 72 horas após a extubação traqueal, utilizando-se a Escala Analógica Visual (EAV) e Escala de Glasgow Modificada (EGM). Analgesia de resgate foi feita com morfina (0,5mg kg-1 IM) em casos do escore de dor ser superior a 50% da EAV e/ou 33% da EGM. Os escores de dor foram inferiores no tratamento TXB de 8 a 60 horas na EAV e de 12 a 60 horas na EGM. A necessidade de suplementação analgésica foi inferior no gruo TXB (2 de 8 animais) em relação ao grupo Controle (7 de 8 animais). Estabilidade cardiorrespiratória foi observada em ambos os tratamentos. A administração preemptiva de toxina botulínica representa uma alternativa viável para o controle da dor pós-mastectomia em cadelas.

Proteases play an indispensable role in medicine, serving both as drug targets and as therapeuticagents. Protease inhibitors are a key component of our antiviral arsenal, and are widely used to combat human immunodeficiency virus (HIV), hepatitis C virus (HCV), and, most recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Nirmatrelvir, an inhibitor of the 3-chymotrypsin like (3CL) protease essential for SARS-CoV-2 replication, was granted emergency use authorization in late 2021, formulated with ritonavir and sold under the brand name Paxlovid. Since then, Paxlovid use has become widespread, raising the possibility that nirmatrelvir resistance could emerge in circulating SARS-CoV-2. It is therefore important to understand how the 3CL protease might mutate to lose nirmatrelvir sensitivity so that circulating variants can be monitored for these mutations and future generations of inhibitors can be designed to prevent cross-resistance. The most widely used therapeutic proteases are the botulinum neurotoxins (BoNTs),which are effective in the treatment of a wide array of movement, pain, and autonomic disorders. These toxins exert their therapeutic effect by cleaving members of the SNARE protein family inside neuronal cytosol, preventing neurotransmitter release. Much work has been dedicated to engineering members of the BoNT family to extend their therapeutic utility, including altering receptor tropism, extending half-life, and modifying protease specificity. While significant progress has been made in each of these areas, the extent to which these proteases can be reprogrammed to target the proteins that cause human disease remains unexplored. In this dissertation, we employ diverse methods to study and direct protease evolutionand to facilitate protein engineering more broadly. To investigate the emergence of nirmatrelvir resistance, we passage SARS-CoV-2 in increasing concentrations of the drug and sequence the 3CL protease gene over time in resistant lineages. We then validate the observed 3CL protease mutations by incorporating them into recombinant SARS-CoV-2 and testing the nirmatrelvir sensitivity of the resulting viruses. We find that the development of nirmatrelvir resistance typically begins with the acquisition of a precursor mutation such as T21I, P252L, or T304I, which confers a low level of resistance and enables strong resistance conferring mutations, such as E166V, to emerge without imposing a significant fitness cost. To explore the programmability of the BoNT proteases, we engineer type E (BoNT/E) to cleave proteins involved in neurodegenerative disease. To accomplish this, we employ targeted mutation based on a structural model as well as the continuous evolution platform known as OrthoRep, selecting for variants that cleave the desired substrates with a circuit that links protease activity to the growth of a Saccharomyces cerevisiae (yeast) cellular chassis. We first use this approach to target ATXN3, the protein whose aggregation causes type 3 spinocerebellar ataxia (SCA3). We then profile the substrate specificities of the BoNT/E variants that emerged during our ATXN3 engineering, identifying patterns that can inform the selection of new targets. Based on one of these patterns, we generate a new BoNT/E variant capable of cleaving TDP-43, a protein implicated in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Finally, to facilitate future protein engineering campaigns, we develop a machine learning pipeline that uses deep mutational scanning (DMS) data to model a protein’s fitness landscape and predict optimized variants with user-defined constraints. We name this pipeline OptiProt, and we demonstrate its utility by applying it to the human chaperone DNAJB6, which rescues cellular toxicity in a yeast model of ALS. In this context, we probe OptiProt’s engineering capabilities with a range of challenges. We first predict hyper-functional variants with up to 50 mutations. We then restore activity to variants harboring one of two known loss-offunction mutations. Finally, to demonstrate the OptiProt’s amenability to complex engineering constraints, we restore activity to a variant harboring a deleterious mutation while simultaneously mutating a set of highly conserved residues. Altogether, the work presented in this dissertation highlights the value of laboratory protease evolution in two clinically relevant applications and provides a tool that can facilitate future work in this field.

INTRODUÇÃO: A paralisia facial de longa duração cursa com déficit funcional e estético, responsáveis por distúrbios psicológicos e prejuízo na qualidade de vida. Mesmo técnicas cirúrgicas modernas de reanimação conseguem restabelecer apenas parcialmente a movimentação emocional e a simetria entre as hemifaces. A toxina botulínica tipo A provoca paralisia muscular flácida reversível, corrigindo desequilíbrios entre músculos agonistas hipoativos e antagonistas relativamente hiperativos. Não há na literatura séries padronizadas com mais de 10 pacientes com paralisia facial tratados por meio da aplicação de toxina botulínica no lado são a fim de obter maior simetria na dinâmica facial na região da boca. MÉTODOS: Este estudo prospectivo teve como objetivo avaliar o efeito do tratamento adjuvante com toxina botulínica no lado são de 25 pacientes com paralisia facial de longa duração, previamente tratados cirurgicamente. O tempo de seguimento foi de 6 meses. Foram métodos de avaliação escala clínica padronizada, grau de satisfação do paciente, Índices de Função Física (IFF) e de Bem-Estar Social (IBES) e eletromiografia de superfície. A dose total de toxina botulínica variou de 15 a 69 U, média 37,9 ± 5,4 U. Dezesseis pacientes apresentaram efeitos adversos com duração média de 14,1 ± 7,3 dias, incluindo dificuldade para articular palavras, beber, mastigar e se adaptar à redução do sorriso. RESULTADOS: 1) Houve redução significante da assimetria entre as hemifaces, de 48,4% após 1 mês e 16,8% aos 6 meses. O ganho de simetria após 1 mês ocorreu em decorrência da diminuição do movimento do lado são, combinada à melhora da avaliação do lado paralisado. Aos 6 meses, com a perda do efeito clínico da toxina botulínica, a nota do lado são voltou a ser semelhante à do pré-tratamento. A redução de assimetria nessa fase ocorreu devido à melhora significante do lado paralisado em relação ao pré-tratamento. 2) A avaliação subjetiva do paciente em relação à simetria apresentou aumento significante, 1 mês e 6 meses em relação ao pré-tratamento. 3) Houve aumento do IFF ao longo do tempo, porém não significante. O IBES apresentou aumento significante aos 6 meses, comparado ao pré-tratamento. 4) Um mês após a aplicação, o efeito da toxina botulínica levou à diminuição significante do potencial de ação do lado não-paralisado. Após 6 meses, o valor voltou a ser semelhante ao do pré-tratamento. CONCLUSÕES: O tratamento proposto permitiu, com técnica minimamente invasiva, obter melhor simetria facial estática, evidenciada pela posição do ângulo da boca, do filtro labial, dos sulcos nasogenianos, do nariz e do supercílio, e melhor simetria dinâmica, principalmente ao sorrir, falar, na exposição dos dentes e na movimentação facial como um todo. Mesmo após a perda do efeito clínico da droga aos 6 meses, houve 18% de melhora da avaliação clínica do lado paralisado em relação ao pré-tratamento, e melhora dos índices de satisfação e qualidade de vida dos pacientes. / INTRODUCTION: Long-standing facial paralysis presents with functional and aesthetic deficits, which are responsible for psychological disturbances and life quality impairment. Even after modern facial reanimation surgical techniques, the emotional movement and the symmetry of the hemifaces is only partially restablished. Botulinum toxin type A causes reversible flacid muscle paralysis, thus correcting imbalances among hypoactive agonists and relatively hyperactive antagonists. There are no standardized series in the literature with more than 10 facial paralysis patients treated with botulinum toxin injection in the non-paralysed side in the mouth area, with the objective of obtaining better dynamic facial symmetry. METHODS: This prospective study had the objective of evaluate, with 6 months follow-up, the effects of the adjuvant treatment using botulinum toxin in the healthy hemiface of 25 patients with long-standing facial paralysis, previously treated surgically. The methods of evaluation were a standardized clinical scale, the patients degree of satisfaction, the Physical Function and Social/well-being Function subscales of the Facial Disability Index and surface electromyography. Total botulinum toxin dose varied from 15 to 69 U, mean 37,9 ± 5,4 U. Sixteen patients presented adverse effects with mean duration time 14,1 ± 7,3 days, including difficulty in speaking, drinking, eating and adapting to the reduced smile. RESULTS: 1) There was significant reduction of facial asymmetry, of 48,4% at 1 month and of 16,8% at 6 months post-treatment. The better symmetry 1 month post-treatment was consequent to reduced movement on the non-paralysed side combined to better evaluation on the paralysed side. At 6 months, the non-paralysed side had similar grading than that of pre-treatment, showing absence of clinical effect of the toxin. At this time, the asymmetry reduction was due to significant increase in the evaluation of the paralysed side in relation to the pre-treatment. 2) Patients satisfaction with facial symmetry showed significant increase, 1 month and 6 months post-treatment. 3) The Physical Function Index increased, but not significantly. The Social/well-being Function Index showed significant increase at 6 months compared to pre-treatment. 4) There was significant decrease in the action potential of the non-paralysed side one month post-injection of the botulinum toxin. After 6 months, the value returned to baseline. CONCLUSIONS: The proposed treatment allowed, with minimally invasive technique, better facial symmetry at rest, evidenced by better mouth, nose and brow position, and on facial movement as a whole, especially when smiling, speaking or exposing teeth. Even after the loss of the clinical effect of the drug at 6 months, there was an 18% increase in the clinical evaluation of the paralysed side in relation to pre-treatment, and increase in the satisfaction and quality of life indexes.