Electroporation-mediated interleukin-10 overexpression in skeletal muscle reduces acute rejection in rat cardiac allografts

OBJECTIVES: Human interleukin 10 (hIL-10) may reduce acute rejection after organ transplantation. Our previous data shows that electroporation-mediated transfer of plasmid DNA to peripheral muscle enhances gene transduction dramatically. This study was designed to investigate the effect of electroporation-mediated overexpression of hIL-10 on acute rejection of cardiac allografts in the rat. METHODS: The study was designed to evaluate the effect of hIL-10 gene transfer on (a) early rejection pattern and (b) graft survival. Gene transfer was achieved by intramuscular (i.m.) injection into the tibialis anterior muscle of Fischer (F344) male recipients followed by electroporation 24 h prior to transplantation. Heterotopic cardiac transplantation was performed from male Brown Norway rat to F344. Four groups were studied (n = 6). Treated animals in groups B1 and B2 received 2.5 microg of pCIK hIL-10 and control animals in groups A1 and A2 distilled water. Graft function was assessed by daily palpation. Animals from group A1 were sacrificed at the cessation of the heart beat of the graft and those in group B1 were sacrificed at day 7; blood was taken for ELISA measurement of hIL-10 and tissue for myeloperoxidase (MPO) measurement and histological assessment. To evaluate graft survival, groups A2 and B2 were sacrificed at cessation of the heart beat of the graft. RESULTS: Histological examination revealed severe rejection (IIIB-IV) in group A1 in contrast to low to moderate rejection (IA-IIIA) in group B1 (p = 0.02). MPO activity was significantly lower in group B1 compared to group A1 (18 +/- 7 vs. 32 +/- 14 mU/mg protein, p = 0.05). Serum hIL-10 levels were 46 +/- 13 pg/ml in group B1 vs. 0 pg/ml in group A1. At day 7 all heart allografts in the treated groups B1 and B2 were beating, whereas they stopped beating at 5 +/- 2 days in groups A1 and A2 vs. 14 +/- 2 days in group B2 (p = 0.0012). CONCLUSIONS: Electroporation-mediated intramuscular overexpression of hIL-10 reduces acute rejection and improves survival of heterotopic heart allografts in rats. This study demonstrates that peripheral overexpression of specific genes in skeletal muscle may reduce acute rejection after whole organ transplantation.

Influence of muscle fiber orientation on water and metabolite relaxation times, magnetization transfer, and visibility in human skeletal muscle

PURPOSE To gain a deeper understanding of the influence of skeletal muscle fiber orientation on metabolite visibility, magnetization transfer from water, and water proton relaxation rates in (1) H MR spectra. METHODS Non-water-suppressed MR spectroscopy was performed in tibialis anterior muscle (TA) of 10 healthy adults, with the TA oriented either parallel or at the magic angle to the 3T field. Spectra were acquired with metabolite-cycled PRESS, and water inversion from 50 to 2510 ms before excitation. Water proton T2 relaxation was sampled with STEAM with echo times from 12 to 272 ms. RESULTS Apparent concentrations of total creatine (tCr), taurine, and trimethylammonium compounds were reduced by 29% to 67% when TA was parallel to B0 . Both tCr peak areas were strongly correlated to the methylene peak splitting. Magnetization transfer rates from water to tCr CH3 were not significantly different between orientations. Water T1 s were similar between orientations, but T2 s were statistically significantly shorter by 1 ms in the parallel orientation (P = 0.002). CONCLUSION Muscle metabolite visibilities in MR spectroscopy and water T2 times depend substantially on muscle fiber orientation relative to B0 . In contrast, magnetization transfer rates appear to depend on muscle composition, rather than fiber orientation. Magn Reson Med, 2015. © 2015 Wiley Periodicals, Inc.

Physiological time structure of the tibialis anterior motor activity during sleep in mice, rats and humans

The validation of rodent models for restless legs syndrome (Willis-Ekbom disease) and periodic limb movements during sleep requires knowledge of physiological limb motor activity during sleep in rodents. This study aimed to determine the physiological time structure of tibialis anterior activity during sleep in mice and rats, and compare it with that of healthy humans. Wild-type mice (n = 9) and rats (n = 8) were instrumented with electrodes for recording the electroencephalogram and electromyogram of neck muscles and both tibialis anterior muscles. Healthy human subjects (31 ± 1 years, n = 21) underwent overnight polysomnography. An algorithm for automatic scoring of tibialis anterior electromyogram events of mice and rats during non-rapid eye movement sleep was developed and validated. Visual scoring assisted by this algorithm had inter-rater sensitivity of 92-95% and false-positive rates of 13-19% in mice and rats. The distribution of the time intervals between consecutive tibialis anterior electromyogram events during non-rapid eye movement sleep had a single peak extending up to 10 s in mice, rats and human subjects. The tibialis anterior electromyogram events separated by intervals <10 s mainly occurred in series of two-three events, their occurrence rate in humans being lower than in mice and similar to that in rats. In conclusion, this study proposes reliable rules for scoring tibialis anterior electromyogram events during non-rapid eye movement sleep in mice and rats, demonstrating that their physiological time structure is similar to that of healthy young human subjects. These results strengthen the basis for translational rodent models of periodic limb movements during sleep and restless legs syndrome/Willis-Ekbom disease.

Influence of fetal tissue transplant on the morphology of the neuromuscular junctions of tibialis anterior and medial gastrocnemius following spinal transection in the rat

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Influence of fetal tissue transplant on the morphology of the neuromuscular junctions of tibialis anterior and medial gastrocnemius following spinal transection in the rat

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Spinal cord injury therapy through active learning

Therapy employing epidural electrostimulation holds great potential for improving therapy for patients with spinal cord injury (SCI) (Harkema et al., 2011). Further promising results from combined therapies using electrostimulation have also been recently obtained (e.g., van den Brand et al., 2012). The devices being developed to deliver the stimulation are highly flexible, capable of delivering any individual stimulus among a combinatorially large set of stimuli (Gad et al., 2013). While this extreme flexibility is very useful for ensuring that the device can deliver an appropriate stimulus, the challenge of choosing good stimuli is quite substantial, even for expert human experimenters. To develop a fully implantable, autonomous device which can provide useful therapy, it is necessary to design an algorithmic method for choosing the stimulus parameters. Such a method can be used in a clinical setting, by caregivers who are not experts in the neurostimulator's use, and to allow the system to adapt autonomously between visits to the clinic. To create such an algorithm, this dissertation pursues the general class of active learning algorithms that includes Gaussian Process Upper Confidence Bound (GP-UCB, Srinivas et al., 2010), developing the Gaussian Process Batch Upper Confidence Bound (GP-BUCB, Desautels et al., 2012) and Gaussian Process Adaptive Upper Confidence Bound (GP-AUCB) algorithms. This dissertation develops new theoretical bounds for the performance of these and similar algorithms, empirically assesses these algorithms against a number of competitors in simulation, and applies a variant of the GP-BUCB algorithm in closed-loop to control SCI therapy via epidural electrostimulation in four live rats. The algorithm was tasked with maximizing the amplitude of evoked potentials in the rats' left tibialis anterior muscle. These experiments show that the algorithm is capable of directing these experiments sensibly, finding effective stimuli in all four animals. Further, in direct competition with an expert human experimenter, the algorithm produced superior performance in terms of average reward and comparable or superior performance in terms of maximum reward. These results indicate that variants of GP-BUCB may be suitable for autonomously directing SCI therapy.

Rôle du stress oxydant en période néonatale dans l'hypertension artérielle et la dysfonction vasculaire et métabolique de l'adulte

Thèse réalisée dans le cadre d'une cotutelle entre l'Université de Montréal et l'Université d'Auvergne en France

Análise da fadiga muscular localizada em atletas e sedentários através de parâmetros de freqüência do sinal eletromiográfico

Embora a análise no domínio da freqüência do sinal eletromiográfico (EMG) seja empregada na caracterização do processo de fadiga muscular localizada, sua aplicação, especificamente a da freqüência mediana (Fmed), é pouco explorada no âmbito esportivo. O objetivo do presente estudo foi verificar a viabilidade da aplicação do sinal EMG, através de sua análise no domínio da freqüência, como parâmetro para determinação e diferenciação no comportamento da fadiga muscular localizada. Dois grupos de sujeitos, um caracterizado como atletas (n =12) e outro como sedentários (n =12), foram submetidos a análises baseadas em procedimentos executados em três diferentes situações experimentais, todos envolvendo a modalidade de exercício isométrico: i) teste máximo para determinação da contração isométrica voluntária máxima (CIVM); ii) teste de fadiga, sustentado por 35 seg. a 80% da CIVM; iii) teste de recuperação, sustentado por 10 seg. a 80% da CIVM; neste ultimo foi monitorado o comportamento da Fmed nos três primeiros (Fmedi) e três últimos segundos (Fmedf) do sinal EMG no músculo tibial anterior durante o teste de fadiga. Durante os 10 segundos do teste de recuperação foi calculada a Fmed referente a todo o período (Fmedr). parâmetro utilizado no cálculo do índice de recuperação muscular (IRM). Os resultados apontam que a Fmedf apresentou valor menor em relação à Fmedi em ambos os grupos (p < 0,05). Quando comparado com o grupo de sedentários, o grupo de atletas apresentou valores maiores de Fmedi e Fmedf (p < 0,05). O valor médio e desvio-padrão do IRM para o grupo de atletas foram de 62,1% ± 28,7 e, para o grupo de sedentários, de 55,2% ± 27,8 (p > 0,05). Dessa forma, os resultados apresentados neste estudo permitem inferir a viabilidade na aplicação de parâmetros no domínio da freqüência do sinal EMG para a determinação e diferenciação do comportamento da fadiga muscular localizada.

Análise qualitativa dos efeitos da sonoforese com Arnica montana sobre o processo inflamatório agudo do músculo esquelético de ratos

Este estudo visou verificar o efeito da sonoforese com Arnica montana sobre a fase inflamatória aguda de uma lesão muscular. Para isso, 40 ratos Wistar machos, lesados cirurgicamente, foram divididos em 4 grupos: controle (C), 10 ratos lesados e não tratados; grupo ultra-som (US), 10 lesados, tratados com US; grupo ultra-som com arnica (US+A), 10 ratos lesados, tratados com sonoforese de gel de arnica; grupo arnica (A), 10 ratos lesados, tratados com massagem de gel de arnica. O tratamento dos três grupos foi iniciado 24h após a lesão, sendo aplicado uma vez ao dia durante 3 minutos, por três dias. Quatro dias após a lesão, os animais foram sacrificados e o terço médio do músculo tibial anterior lesado foi removido e tratado histologicamente. Os resultados da análise qualitativa mostram que, no grupo C, formou-se um intenso infiltrado de células inflamatórias no espaço intersticial e um processo de regeneração apenas iniciado. Nos grupos US e US+A foi detectado um avançado processo inflamatório, com tecido conjuntivo mais organizado e consistente. No grupo A foi detectada diminuição no número de células inflamatórias e uma desorganização em sua disposição, o que poderia levar a um atraso no processo de regeneração. Conclui-se que os grupos que receberam a aplicação do ultra-som e ultra-som com arnica apresentaram semelhante aceleração do processo inflamatório agudo, sugerindo ineficácia da sonoforese quando comparada à aplicação de apenas ultra-som.

Effects of phonophoresis with Arnica montana onto acute inflammatory process in rat skeletal muscles: An experimental study

This study aimed at verifying the effects of phonophoresis associated with Arnica montana on the acute phase of an inflammatory muscle lesion. Forty Wistar male rats (300 +/- 50 g), of which the Tibialis Anterior muscle was surgically lesioned, were divided into four groups (n = 10 each): control group received no treatment; the ultrasound group (US) was treated in pulsed mode with 1-MHz frequency, 0.5 W/cm(2) intensity (spatial and temporal average - SATA), duty cycle of 1: 2 (2 ms on, 4 ms off, 50%), time of application 3 min per session, one session per day, for 3 days; the phonophoresis or ultrasound plus arnica (US+A) group was treated with arnica with the same US parameters plus arnica gel; and the arnica group (A) was submitted to massage with arnica gel, also for 3 min, once a day, for 3 days. Treatment started 24 h after the surgical lesion. on the 4th day after lesion creation, animals were sacrificed and sections of the lesioned, inflamed muscle were removed for quantitative (mononuclear and polymorphonuclear cell count) and qualitative histological analysis. Collected data from the 4 groups were statistically analyzed and the significance level set at p < 0.05. Results show higher mononuclear cell density in all three treated groups with no significant difference between them, but values were significantly different (p < 0.0001) when compared to control group's. As to polymorphonuclear cell density, significant differences were found between control group (p = 0.0134) and US, US+A and A groups; the arnica group presented lesser density of polymorphonuclear cells when compared (p = 0.0134) to the other groups. No significant difference was found between US and US+A groups. While the massage with arnica gel proved to be an effective anti-inflammatory on acute muscle lesion in topic use, these results point to ineffectiveness of Arnica montana phonophoresis, US having seemingly checked or minimized its anti-inflammatory effect. (C) 2008 Elsevier B. V. All rights reserved.

Static balance and function in children with cerebral palsy submitted to neuromuscular block and neuromuscular electrical stimulation: Study protocol for prospective, randomized, controlled trial

Background: The use of botulinum toxin A (BT-A) for the treatment of lower limb spasticity is common in children with cerebral palsy (CP). Following the administration of BT-A, physical therapy plays a fundamental role in potentiating the functionality of the child. The balance deficit found in children with CP is mainly caused by muscle imbalance (spastic agonist and weak antagonist). Neuromuscular electrical stimulation (NMES) is a promising therapeutic modality for muscle strengthening in this population. The aim of the present study is to describe a protocol for a study aimed at analyzing the effects of NMES on dorsiflexors combined with physical therapy on static and functional balance in children with CP submitted to BT-A.Methods/Design: Protocol for a prospective, randomized, controlled trial with a blinded evaluator. Eligible participants will be children with cerebral palsy (Levels I, II and III of the Gross Motor Function Classification System) between five and 12 years of age, with independent gait with or without a gait-assistance device. All participants will receive BT-A in the lower limbs (triceps surae). The children will then be randomly allocated for either treatment with motor physical therapy combined with NMES on the tibialis anterior or motor physical therapy alone. The participants will be evaluated on three occasions: 1) one week prior to the administration of BT-A; 2) one week after the administration of BT-A; and 3) four months after the administration of BT-A (end of intervention). Spasticity will be assessed by the Modified Ashworth Scale and Modified Tardieu Scale. Static balance will be assessed using the Medicapteurs Fusyo pressure platform and functional balance will be assessed using the Berg Balance Scale.Discussion: The aim of this protocol study is to describe the methodology of a randomized, controlled, clinical trial comparing the effect of motor physical therapy combined with NMES on the tibialis anterior muscle or motor physical therapy alone on static and functional balance in children with CP submitted to BT-A in the lower limbs. This study describes the background, hypotheses, methodology of the procedures and measurement of the results.

Fator de crescimento nervoso em cola de fibrina no reparo término-lateral de nervos em ratos

Purpose: To determine the effects of end-to-side nerve repair performed only with fibrin glue containing nerve growth in rats. Methods: Seventy two Wistar rats were divided into six equal groups: group A was not submitted to nerve section; group B was submitted to nerve fibular section only. The others groups had the nerve fibular sectioned and then repaired in the lateral surface of an intact tibial nerve, with different procedures: group C: ETS with sutures; group D: ETS with sutures and NGF; group E: ETS with FG only; group F: ETS with FG containing NGF. The motor function was accompanied and the tibial muscle mass, the number and diameter of muscular fibers and regenerated axons were measured. Results: All the analyzed variables did not show any differences among the four operated groups (p>0.05), which were statistically superior to group B (p<0.05), but inferior to group A (p>0.05). Conclusion: The end-to-side nerve repair presented the same recovery pattern, independent from the repair used, showing that the addition of nerve growth factor in fibrin glue was not enough for the results potentiating.

Motor readiness and joint torque production in lower limbs of older women fallers and non-fallers

This study aimed to evaluate the motor response time and ability to develop joint torque at the knee and ankle in older women with and without a history of falls, in addition to investigating the effect of aging on these capacities. We assessed 18 young females, 21 older female fallers and 22 older female non-fallers. The peak torque, rate of torque development, rate of electromyography (EMG) rise, reaction time, premotor time and motor time were obtained through a dynamometric assessment and simultaneous electromyography. Surface EMGs of the rectus femoris (RF), vastus lateralis (VL), biceps femoris (BF), gastrocnemius lateralis (GL) and tibialis anterior (TA) muscles were recorded. Knee extension and flexion peak torques were lower in older fallers than in non-fallers. Knee extension and flexion and ankle plantarflexion and dorsiflexion peak torques were lower in both older groups than in the younger group. The rate of EMG rise of the BF and the motor time of the TA were lower and higher, respectively, in older fallers than in the younger adults. The time to reach peak torque in knee extension/flexion and ankle plantarflexion/dorsiflexion and the motor times of the RF, VL, BF and GL were higher in both older groups than in the younger groups. The motor time of the TA during ankle dorsiflexion and the knee extension peak torque were the major predictors of falls in older women, accounting for approximately 28% of the number of falls. Thus, these results further reveal the biomechanical parameters that affect the risk of falls and provide initial findings to support the prescription of exercises in fall prevention programs. © 2013 Elsevier Ltd.