Forelimb Force Research Articles

Heavy metal pollution alters reproductive performance and mate choice in an anuran, Strauchbufo raddei

Understanding the effects of pollution on reproductive performance and sexual selection is crucial for the conservation of biodiversity in an increasingly polluted world. The present study focused on the effect of environmental heavy metal pollution on sexually selected traits, including morphological characteristics and acoustic parameters, as well as mate choice in Strauchbufo raddei, an anuran species widely distributed in Northern China. The results showed that male courtship signals, including forelimb length, forelimb force, and advertisement calls, have evolved under the pressure of heavy metal pollution in young S. raddei. In addition, the breeding age was lower in the polluted areas, and younger individuals had more mating opportunities. However, males with heightened reproductive performance did not show the expected higher individual quality. The current study suggests that exposure to heavy metal pollution can induce stress in males, altering reproductive performance and further disrupting mate choice.

Growth hormone secretagogues modulate inflammation and fibrosis in mdx mouse model of Duchenne muscular dystrophy.

Growth hormone secretagogues (GHSs) exert multiple actions, being able to activate GHS-receptor 1a, control inflammation and metabolism, to enhance GH/insulin-like growth factor-1 (IGF-1)-mediated myogenesis, and to inhibit angiotensin-converting enzyme. These mechanisms are of interest for potentially targeting multiple steps of pathogenic cascade in Duchenne muscular dystrophy (DMD). Here, we aimed to provide preclinical evidence for potential benefits of GHSs in DMD, via a multidisciplinary in vivo and ex vivo comparison in mdx mice, of two ad hoc synthesized compounds (EP80317 and JMV2894), with a wide but different profile. 4-week-old mdx mice were treated for 8 weeks with EP80317 or JMV2894 (320 µg/kg/d, s.c.). In vivo, both GHSs increased mice forelimb force (recovery score, RS towards WT: 20% for EP80317 and 32% for JMV2894 at week 8). In parallel, GHSs also reduced diaphragm (DIA) and gastrocnemius (GC) ultrasound echodensity, a fibrosis-related parameter (RS: ranging between 26% and 75%). Ex vivo, both drugs ameliorated DIA isometric force and calcium-related indices (e.g., RS: 40% for tetanic force). Histological analysis highlighted a relevant reduction of fibrosis in GC and DIA muscles of treated mice, paralleled by a decrease in gene expression of TGF-β1 and Col1a1. Also, decreased levels of pro-inflammatory genes (IL-6, CD68), accompanied by an increment in Sirt-1, PGC-1α and MEF2c expression, were observed in response to treatments, suggesting an overall improvement of myofiber metabolism. No detectable transcript levels of GHS receptor-1a, nor an increase of circulating IGF-1 were found, suggesting the presence of a novel receptor-independent mechanism in skeletal muscle. Preliminary docking studies revealed a potential binding capability of JMV2894 on metalloproteases involved in extracellular matrix remodeling and cytokine production, such as ADAMTS-5 and MMP-9, overactivated in DMD. Our results support the interest of GHSs as modulators of pathology progression in mdx mice, disclosing a direct anti-fibrotic action that may prove beneficial to contrast pathological remodeling.

Relationship between Thoroughbred workloads in racing and the fatigue life of equine subchondral bone

Fatigue life (FL) is the number of cycles of load sustained by a material before failure, and is dependent on the load magnitude. For athletes, ‘cycles’ translates to number of strides, with load proportional to speed. To improve previous investigations estimating workload from distance, we used speed (m/s, x) per stride collected using 5 Hz GPS/800 Hz accelerometer sensors as a proxy for limb load to investigate factors associated with FL in a Thoroughbred race start model over 25,234 race starts, using a combination of mathematical and regression modelling. Fore-limb vertical force (NKg-1) was estimated using a published equation: Vertical force = 2.778 + 2.1376x − 0.0535x2. Joint load (σ) was estimated based on the vertical force, scaled according to the maximum speed and defined experimental loads for the expected variation in load distribution across a joint surface (54-90 MPa). Percentage FL (%FL) was estimated using a published equation for cycles to failure (Nf) summed across each race start: Nf = 10(σ-134.2)/−14.1. Multivariable mixed-effects linear regression models were generated on %FL, adjusting for horse-level clustering, presented as coefficients; 95%CI. Scaled to the highest joint load, individual starts accrued a mean of 9.34%FL (sd. 1.64). Older age (coef. 0.03; 0.002–0.04), longer race-distances (non-linear power transformed), and firmer track surfaces (ref. Heavy 10: Good 3 coef. 2.37; 2.26–2.48) were associated with greater %FL, and males accrued less than females (p < 0.01). Most variables associated with %FL are reported risk factors for injury. Monitoring strides in racehorses may therefore allow identification of horses at risk, enabling early detection of injury.

Forelimb force direction and magnitude independently controlled by spinal modules in the macaque

Modular organization of the spinal motor system is thought to reduce the cognitive complexity of simultaneously controlling the large number of muscles and joints in the human body. Although modular organization has been confirmed in the hindlimb control system of several animal species, it has yet to be established in the forelimb motor system or in primates. Expanding upon experiments originally performed in the frog lumbar spinal cord, we examined whether costimulation of two sites in the macaque monkey cervical spinal cord results in motor activity that is a simple linear sum of the responses evoked by stimulating each site individually. Similar to previous observations in the frog and rodent hindlimb, our analysis revealed that in most cases (77% of all pairs) the directions of the force fields elicited by costimulation were highly similar to those predicted by the simple linear sum of those elicited by stimulating each site individually. A comparable simple summation of electromyography (EMG) output, especially in the proximal muscles, suggested that this linear summation of force field direction was produced by a spinal neural mechanism whereby the forelimb motor output recruited by costimulation was also summed linearly. We further found that the force field magnitudes exhibited supralinear (amplified) summation, which was also observed in the EMG output of distal forelimb muscles, implying a novel feature of primate forelimb control. Overall, our observations support the idea that complex movements in the primate forelimb control system are made possible by flexibly combined spinal motor modules.

P.321Diaphragm echodensity and function in mdx mice by non-invasive ultrasonography: validation and correlation with in vivo and ex vivo functional readouts

In the mdx mouse model of Duchenne muscular dystrophy (DMD), diaphragm (DIA) alterations occur at early age, with a progression which closely resembles patients' condition. High frequency ultrasound has been recently proposed as a longitudinal, non-invasive technique to monitor the progression of mdx DIA dysfunction. At present, data available are still limited, particularly in mdx mice at early disease stages. Moreover, no evidence has ever been provided about the usefulness of ultrasonography to measure echodensity, as an index of contractile tissue loss in mdx DIA. In light of this, and considering the importance assumed by an independent validation of new experimental procedures in preclinical studies on DMD, our study was aimed to perform an ultrasonography evaluation of DIA amplitude and echodensity, in 3- and 6-month-old mdx mice (the age window typically used for pharmacological studies), compared to wild type (wt). In parallel, in vivo (forelimb force, fatigability) and ex vivo (DIA contractile recordings, TGF-β1 levels) dystrophy-relevant outcomes were assessed. Ultrasonography showed a significant decrease of DIA amplitude in 3- and 6-month-old mdx mice compared to wt, which correlated with the isometric force reduction observed ex vivo. An age-dependent increase of mdx DIA echodensity was disclosed; this paralleled the high levels of muscular pro-fibrotic TGF-β1. DIA from 6-month-old mdx mice also showed a consistent force drop after eccentric contraction and a change in stiffness compared to wt, likely in relation to an increased fibrosis. Notably, no alteration of left ventricular function was found in mdx hearts, supporting the view that respiratory complications occur independently from cardiac ones. Our findings corroborate the usefulness of DIA ultrasonography as a tool to evaluate mdx mouse pathology progression since early stages, and this possibility opens important new perspectives to improve translational research on DMD (Supported by PRIN-MIUR n. 20108YB5W3_004).

Sagittal plane fore hoof unevenness is associated with fore and hindlimb asymmetrical force vectors in the sagittal and frontal planes.

Asymmetry in forelimb dorsal hoof wall angles, termed unevenness, is associated with forelimb gait asymmetries, but compensatory mechanisms and out of plane ground reaction forces (GRFs) due to unevenness have yet to be documented. The aim of this study was therefore to investigate the effects of fore hoof unevenness on contralateral fore and hind limb force vectors patterns, in both sagittal and frontal planes. A group of n = 34 riding horses were classified into four groups: hoof angle difference of more than 1.5 degrees (UNEVEN; n = 27), including higher left fore (HIGH-LF; n = 12), higher right fore (HIGH-RF; n = 15), and hoof angle difference of less than 1.5 degrees (EVEN; n = 7). Three dimensional ground reaction forces GRFs were collected during trotting. GRF summary vectors representing the magnitude (VecMag) and angular direction (VecAng) of the entire stance phase in the sagittal and the frontal plane were calculated. The effects of unevenness on GRF production were explored using linear regression, repeated measures ANOVA and statistical parametric mapping (SPM) with significance at (P<0.05). In all uneven groups, increasing unevenness affected sagittal VecAng values in the forelimbs, with more propulsive GRF in the high hoof. In the HIGH-RF group, medial GRFs were also found in the high RF hoof compared to lateral GRFs in the low LF hoof (RF VecAng: 0.97±1.64 (deg); LF VecAng: -0.64±1.19 (deg); P<0.05). In both HIGH groups, compensatory associations to increasing unevenness were only found in the RH, but also a significantly greater lateral VecAng was found in the LH (P<0.05) compared to the RH limb. No significant differences (P>0.05) were found between hindlimb pairs in the EVEN group. Unbalanced sagittal and increased frontal plane GRFs in uneven horses suggest that they have greater locomotory challenges, as the equine musculoskeletal system is not constructed to withstand movement and loading in the frontal plane as effectively as it is in the sagittal plane.

Growth hormone secretagogues prevent dysregulation of skeletal muscle calcium homeostasis in a rat model of cisplatin-induced cachexia.

BackgroundCachexia is a wasting condition associated with cancer types and, at the same time, is a serious and dose‐limiting side effect of cancer chemotherapy. Skeletal muscle loss is one of the main characteristics of cachexia that significantly contributes to the functional muscle impairment. Calcium‐dependent signaling pathways are believed to play an important role in skeletal muscle decline observed in cachexia, but whether intracellular calcium homeostasis is affected in this situation remains uncertain. Growth hormone secretagogues (GHS), a family of synthetic agonists of ghrelin receptor (GHS‐R1a), are being developed as a therapeutic option for cancer cachexia syndrome; however, the exact mechanism by which GHS interfere with skeletal muscle is not fully understood.MethodsBy a multidisciplinary approach ranging from cytofluorometry and electrophysiology to gene expression and histology, we characterized the calcium homeostasis in fast‐twitch extensor digitorum longus (EDL) muscle of adult rats with cisplatin‐induced cachexia and established the potential beneficial effects of two GHS (hexarelin and JMV2894) at this level. Additionally, in vivo measures of grip strength and of ultrasonography recordings allowed us to evaluate the functional impact of GHS therapeutic intervention.ResultsCisplatin‐treated EDL muscle fibres were characterized by a ~18% significant reduction of the muscle weight and fibre diameter together with an up‐regulation of atrogin1/Murf‐1 genes and a down‐regulation of Pgc1‐a gene, all indexes of muscle atrophy, and by a two‐fold increase in resting intracellular calcium, [Ca2+]i, compared with control rats. Moreover, the amplitude of the calcium transient induced by caffeine or depolarizing high potassium solution as well as the store‐operated calcium entry were ~50% significantly reduced in cisplatin‐treated rats. Calcium homeostasis dysregulation parallels with changes of functional ex vivo (excitability and resting macroscopic conductance) and in vivo (forelimb force and muscle volume) outcomes in cachectic animals. Administration of hexarelin or JMV2894 markedly reduced the cisplatin‐induced alteration of calcium homeostasis by both common as well as drug‐specific mechanisms of action. This effect correlated with muscle function preservation as well as amelioration of various atrophic indexes, thus supporting the functional impact of GHS activity on calcium homeostasis.ConclusionsOur findings provide a direct evidence that a dysregulation of calcium homeostasis plays a key role in cisplatin‐induced model of cachexia gaining insight into the etiopathogenesis of this form of muscle wasting. Furthermore, our demonstration that GHS administration efficaciously prevents cisplatin‐induced calcium homeostasis alteration contributes to elucidate the mechanism of action through which GHS could potentially ameliorate chemotherapy‐associated cachexia.

Effects of Morphine on Behavioral Task Performance in SIV-Infected Rhesus Macaques.

The abuse of opiates such as morphine in synergy with HIV infection not only exacerbates neuropathogenesis but significantly impacts behavioral attributes in HIV infected subjects. Thus, the goal of the current study was to characterize behavioral perturbations in rhesus macaques subjected to chronic morphine and SIV infection. Specifically, we assessed three behavioral tasks: motor skill (MS), forelimb force (FFT) and progressive ratio (PR) tasks. After collecting baseline control data (44weeks) and data during the morphine-only dependency period (26weeks), a subset of animals were productively infected with neurovirulent strains of SIVmac (R71/E17) for an additional 33weeks. A general pattern in the results is that behavioral decline occurred with high CSF viral loads but not necessarily with high plasma viral loads. Compared to saline controls, all treated animals showed significant decreases in performance on all three behavioral tasks during the morphine-only dependency period. During the post infection period, only the morphine plus SIV group showed a significant further decline and this only occurred for the MS task. Taken together, these data demonstrate a clear effect of morphine to produce behavioral deficits and also suggest that morphine can act synergistically with SIV/HIV to exacerbate behavioral deficits.

Robotic Rehabilitator of the Rodent Upper Extremity: A System and Method for Assessing and Training Forelimb Force Production after Neurological Injury.

Rodent models of spinal cord injury are critical for the development of treatments for upper limb motor impairment in humans, but there are few methods for measuring forelimb strength of rodents, an important outcome measure. We developed a novel robotic device--the Robotic Rehabilitator of the Rodent Upper Extremity (RUE)--that requires rats to voluntarily reach for and pull a bar to retrieve a food reward; the resistance of the bar can be programmed. We used RUE to train forelimb strength of 16 rats three times per week for 23 weeks before and 38 weeks after a mild (100 kdyne) unilateral contusion at the cervical level 5 (C5). We measured maximum force produced when RUE movement was unexpectedly blocked. We compared this blocked pulling force (BPF) to weekly measures of forelimb strength obtained with a previous, well-established method: the grip strength meter (GSM). Before injury, BPF was 2.6 times higher (BPF, 444.6 ± 19.1 g; GSM, 168.4 ± 3.1 g) and 4.9 times more variable (p < 0.001) than pulling force measured with the GSM; the two measurement methods were uncorrelated (R(2) = 0.03; p = 0.84). After injury, there was a significant decrease in BPF of 134.35 g ± 14.71 g (p < 0.001). Together, our findings document BPF as a repeatable measure of forelimb force production, sensitive to a mild spinal cord injury, which comes closer to measuring maximum force than the GSM and thus may provide a useful measure for quantifying the effects of treatment in rodent models of SCI.

Abstract T MP24: Vagus Nerve Stimulation Improves Motor Function After Chronic Stroke in Adult Rats

Background: Stroke is the leading cause of serious long-term disability. Currently, there is no effective treatment for chronic stroke patients. Neuroplasticity within motor circuitry is believed to support recovery of function after stroke. We have developed a method using vagus nerve stimulation (VNS) paired with motor training to drive robust, specific plasticity in the motor cortex. Our recent studies indicated that VNS paired with rehab training significantly enhances recovery of forelimb function after cortical ischemic stroke. To further the translation potential of our therapy, we accessed the hypothesis that delivering VNS paired with rehab may improve functional recovery in rats that demonstrated chronic forelimb impairments. Methods: All female Sprague Dawley rats were trained on the Isometric Pull Task, which quantifiably measures forelimb force generation. Rats that achieved 5 consecutive days of over 85% hit rate on this task were given a unilateral cortical-subcortical ischemic lesion via injections of a vasoconstrictive peptide, endothelin-1. Following the lesion, rats returned to their home cage, and did not begin rehab training until 5 weeks post-lesion. Upon return, post-lesion forelimb impairment was accessed with the same task parameters used during pre-lesion training, which allowed for a direct comparison of performance. Rats were assigned to balanced treatment groups based on post-lesion baseline hit rate. Treatment groups consisted of VNS delivered during rehab training (Paired VNS; n=10), VNS delivered two hours after rehab training (Delayed VNS; n=10), and rehab training without VNS (Rehab; n=10). Results: At five weeks post-lesion, the unilateral ischemic insult significantly worsened performance in all three groups compared to pre-lesion (Paired VNS: 29.8 ± 5.7%, paired t-test, P &lt; 0.001; Delayed VNS: 24.6 ± 2.7%, P &lt; 0.001; Rehab: 30.4 ± 4.7%, P &lt; 0.001). Following our therapy, the Paired VNS group demonstrated significantly better performance than both control groups (Paired VNS: 81.6 ± 2.3%, P &lt; 0.01; Delayed VNS: 53.2 ± 6.5%, P &lt; 0.01; Rehab: 49.8 ± 6.8%, P &lt; 0.01). Conclusion: Our results indicate that VNS paired with rehab training can further enhance recovery of forelimb function in chronically impaired rats.

G.P.90: Effects of S48168/Arm210, a new rycal® compound, on pathology related signs of exercised dystrophic mdx mouse

The progressive myofiber degeneration in Duchenne muscular dystrophy (DMD) is caused by the complex cascade of events triggered by the absence of dystrophin. Studies in the mdx mouse model of DMD have shown that pathology-related post-translational modifications of the Ryanodine receptor subtype 1 (RyR1) result in the dissociation of the stabilizing subunit calstabin1, leading to leaky channels and altered Ca2+homeostasis, a key event in DMD (Bellinger, Nat Med 2009). S48168, also known as ARM210, is a small molecule stabilizer of the calstabin-RyR channel complex that prevents Ca 2+ leak without altering RyR1 post-translational modifications. We assessed the effects of 4weeks oral administration of S48168 (10 and 50mg/kg/day) on treadmill-exercised mdx mice, starting at 4–5weeks of age. A functional improvement was observed in vivo in S48168 -treated mdx mice, with a 50% recovery score of maximal forelimb force at 50mg/kg. Both doses prevented the decline in running performance, observed in vehicle-treated mdx mice, as assessed in an exhaustion test. Ex vivo assessment showed a dose-dependent improvement of diaphragm specific force, with a significant 30% increase of maximal force (100–140Hz) at 50mg/kg, while minor, if any, effects were observed in hind-limb EDL muscle. However, and consistently with the S48168 mechanism of action, the mechanical threshold EDL myofibers, an index of calcium handling during contraction, was significantly shifted toward the wild type (wt) values at the highest dose. In parallel, the resting cytosolic calcium level in FDB myofibers was significantly reduced by 25% in the S48168-treated mice. S48168 did not lower CK or LDH plasma levels. However a reduction of damaged area was detected in diaphragm and in gastrocnemius muscle of 50mg/kg treated mdx mice. Dose-proportional increases in plasma and muscle drug content were also observed, without any signs of toxicity. These results support S48168 as a potential therapy for DMD.

Vagus nerve stimulation during rehabilitative training improves forelimb strength following ischemic stroke

Upper limb impairment is a common debilitating consequence of ischemic stroke. Physical rehabilitation after stroke enhances neuroplasticity and improves limb function, but does not typically restore normal movement. We have recently developed a novel method that uses vagus nerve stimulation (VNS) paired with forelimb movements to drive specific, long-lasting map plasticity in rat primary motor cortex. Here we report that VNS paired with rehabilitative training can enhance recovery of forelimb force generation following infarction of primary motor cortex in rats. Quantitative measures of forelimb function returned to pre-lesion levels when VNS was delivered during rehab training. Intensive rehab training without VNS failed to restore function back to pre-lesion levels. Animals that received VNS during rehab improved twice as much as rats that received the same rehabilitation without VNS. VNS delivered during physical rehabilitation represents a novel method that may provide long-lasting benefits towards stroke recovery.

Dynamic motor compensations with permanent, focal loss of forelimb force after cervical spinal cord injury.

Incomplete cervical lesion is the most common type of human spinal cord injury (SCI) and causes permanent paresis of arm muscles, a phenomenon still incompletely understood in physiopathological and neuroanatomical terms. We performed spinal cord hemisection in adult rats at the caudal part of the segment C6, just rostral to the bulk of triceps brachii motoneurons, and analyzed the forces and kinematics of locomotion up to 4 months postlesion to determine the nature of motor function loss and recovery. A dramatic (50%), immediate and permanent loss of extensor force occurred in the forelimb but not in the hind limb of the injured side, accompanied by elbow and wrist kinematic impairments and early adaptations of whole-body movements that initially compensated the balance but changed continuously over the follow-up period to allow effective locomotion. Overuse of both contralateral legs and ipsilateral hind leg was evidenced since 5 days postlesion. Ipsilateral foreleg deficits resulted mainly from interruption of axons that innervate the spinal cord segments caudal to the lesion, because chronic loss (about 35%) of synapses was detected at C7 while only 14% of triceps braquii motoneurons died, as assessed by synaptophysin immunohistochemistry and retrograde neural tracing, respectively. We also found a large pool of propriospinal neurons projecting from C2-C5 to C7 in normal rats, with topographical features similar to the propriospinal premotoneuronal system of cats and primates. Thus, concurrent axotomy at C6 of brain descending axons and cervical propriospinal axons likely hampered spontaneous recovery of the focal neurological impairments.

The isometric pull task: A novel automated method for quantifying forelimb force generation in rats

Reach-to-grasp tasks are commonly used to assess forelimb function in rodent models. While these tasks have been useful for investigating several facets of forelimb function, they are typically labor-intensive and do not directly quantify physiological parameters. Here we describe the isometric pull task, a novel method to measure forelimb strength and function in rats. Animals were trained to reach outside the cage, grasp a handle attached to a stationary force transducer, and pull with a predetermined amount of force to receive a food reward. This task provides quantitative data on operant forelimb force generation. Multiple parameters can be measured with a high degree of accuracy, including force, success rate, pull attempts, and latency to maximal force. The task is fully automated, allowing a single experimenter to test multiple animals simultaneously with usually more than 300 trials per day, providing more statistical power than most other forelimb motor tasks. We demonstrate that an ischemic lesion in primary motor cortex yields robust deficits in all forelimb function parameters measured with this method. The isometric pull task is a significant advance in operant conditioning systems designed to automate the measurement of multiple facets of forelimb function and assess deficits in rodent models of brain damage and motor dysfunction.

Exposure to hypergravity during specific developmental periods differentially affects metabolism and vestibular reactions in adult C57BL /6j mice

The development of the posturo-motor control of movement is conditioned by Earth's gravity. Missing or altered gravity during the critical periods of development delays development and induces durable changes in the vestibular, cerebellar, or muscular structures, but these are not consistently mirrored at a functional level. The differences in the time schedule of vestibular and motor development could contribute to this inconstancy. To investigate the influence of gravity on the development of vestibular and locomotor functions, we analysed the performance of adult mice subjected to hypergravity during the time covering either the vestibular or locomotor development. The mice were centrifuged at 2 g from embryonic day (E) 0 to postnatal day (P) 10 (PRE), from P10 to P30 (POST), from E0 to P30 (FULL), and from E7 to P21. Their muscular force, anxiety level, vestibular reactions, and aerobic capacity during treadmill training were then evaluated at the age of 2 and 6 months. The performance of young adults varied in relation to the period of exposure to hypergravity. The mice that acquired locomotion in hypergravity (POST and FULL) showed a lower forelimb force and delayed vestibular reactions. The mice centrifuged from conception to P10 (PRE) showed a higher aerobic capacity during treadmill training. The differences in muscular force and vestibular reactions regressed with age, but the metabolic changes persisted. These results confirmed that early exposure to hypergravity induces qualitative changes depending on the period of exposure. They validated, at a functional level, the existence of several critical periods for adaptation to gravity.

Young and middle-aged rats exhibit isometric forelimb force control deficits in a model of early-stage Parkinson's disease

Deficits in manual motor control often accompany the early stages of Parkinson's disease (PD), and are often revealed through isometric force tasks. In order to determine whether similar deficits occur in a rat model of early-stage PD, young (8 months) and middle-aged (18 months) rats were trained to produce sustained press-hold-release isometric forelimb responses that allowed for analyses of force output and spectral analysis of forelimb stability and tremor. Rats then received a 6-hydroxydopamine (6-OHDA) infusion into the striatum contralateral to the trained forelimb and were tested for 4 weeks post-lesion. The resulting partial striatal dopamine depletions (which at 41 ± 12% and 43 ± 6% in young and middle-aged rats, respectively, did not differ between the two groups) resulted in isometric forelimb deficits. Specifically, rats exhibited significantly diminished force stability and increased high frequency (10–25 Hz) tremor, indicating potential postural disturbances and increased postural tremor, respectively. Durations of press-hold-release bouts were also increased post-lesion, suggesting difficulty in task disengagement. Despite pre-lesion differences in some of the force measures, the effects of partial nigrostriatal DA depletion did not differ between the two age groups. These results support the use of the press-while-licking task in preclinical studies modeling isometric force control deficits in PD.

Role of tumour necrosis factor α, but not of cyclo‐oxygenase‐2‐derived eicosanoids, on functional and morphological indices of dystrophic progression in mdx mice: a pharmacological approach

The role of tumour necrosis factor (TNF)-alpha or cyclo-oxygenase-2 (COX-2) eicosanoids in dystrophinopathies has been evaluated by chronically treating (4-8 weeks) adult dystrophic mdx mice with the anti-TNF-alpha etanercept (0.5 mg/kg) or the COX-2 inhibitor meloxicam (0.2 mg/kg). Throughout the treatment period the mdx mice underwent a protocol of exercise on treadmill in order to worsen the pathology progression; gastrocnemious muscles from exercised mdx mice showed an intense staining for TNF-alpha by immunohistochemistry. In vivo, etanercept, but not meloxicam, contrasted the exercise-induced forelimb force drop. Electrophysiological recordings ex vivo, showed that etanercept counteracted the decrease in chloride channel function (gCl), a functional index of myofibre damage, in both diaphragm and extensor digitorum longus (EDL) muscle, meloxicam being effective only in EDL muscle. None of the drugs ameliorated calcium homeostasis detected by electrophysiology and/or spectrofluorimetry. Etanercept, more than meloxicam, effectively reduced plasma creatine kinase (CK). Etanercept-treated muscles showed a reduction of connective tissue area and of pro-fibrotic cytokine TGF-beta1 vs. untreated ones; however, the histological profile was weakly ameliorated. In order to better evaluate the impact of etanercept treatment on histology, a 4-week treatment was performed on 2-week-old mdx mice, so to match the first spontaneous degeneration cycle. The histology profile of gastrocnemious was significantly improved with a reduction of degenerating area; however, CK levels were only slightly lower. The present results support a key role of TNF-alpha, but not of COX-2 products, in different phases of dystrophic progression. Anti-TNF-alpha drugs may be useful in combined therapies for Duchenne patients.

Measuring forelimb force control and movement in Fischer 344/Brown Norway rats: effects of age and lorazepam

The purpose of this study was to measure forelimb force control and movement kinetics in rats, as they are affected by normal aging and the benzodiazepine lorazepam. Young (6 months), middle-aged (18 months), and aged (24 months) rats were trained to emit discrete forelimb responses on an isometric force disk within a 20-25 g force band for water reinforcement. Dependent variables included number of responses, percentage of reinforced responses, peak response forces, and inter-response times. Inter-response times were divided into two categories: inter-response times <0.5 s (reflecting rapid, discrete forelimb responses) and inter-response times 4-8 s (reflecting movement sequences). Aged rats exhibited no apparent deficits in forelimb force control. Although older rats emitted fewer responses than younger rats, their response accuracy was greater. Peak forces did not differ among the groups. Both categories of inter-response times were slower in the aged group, reflecting slowed discrete movements and movement sequencing. Lorazepam increased the number of responses and peak forces, decreased response accuracy, and lengthened inter-response times within the 4-8 s range (but not the <0.5 s range) in all age groups. The results suggest that movement sequences may be more sensitive to the effects of acute benzodiazepines than rapid discrete movements.

First evaluation of the potential effectiveness in muscular dystrophy of a novel chimeric compound, BN 82270, acting as calpain-inhibitor and anti-oxidant

BN 82270 is a membrane-permeable prodrug of a chimeric compound (BN 82204) dually acting as calpain inhibitor and anti-oxidant. Acute in vivo injection of dystrophic mdx mice (30 mg/kg, s.c.) fully counteracted calpain overactivity in diaphragm. A chronic 4–6 weeks administration significantly prevented in vivo the fore limb force drop occurring in mdx mice exercised on treadmill. Ex vivo electrophysiological recordings showed that BN 82270 treatment contrasted the decrease in chloride channel function (gCl) in diaphragm, an index of spontaneous degeneration, while it was less effective on both exercise-impaired gCl and calcium-dependent mechanical threshold of the hind limb extensor digitorum longus (EDL) muscle fibres. The BN 82270 treated mdx mice showed a marked reduction of plasma creatine kinase and of the pro-fibrotic cytokine TGF-β1 in both hind limb muscles and diaphragm; however, the histopathological profile of gastrocnemious muscle was poorly ameliorated. In hind limb muscles of treated mice, the active form was detected by HPLC in the low therapeutic concentration range. In vitro exposure to 100 μM BN 82270 led to higher active form in diaphragm than in EDL muscle. This is the first demonstration that this class of chimeric compounds, dually targeting pathology-related events, exerts beneficial effects in muscular dystrophy. The drug/prodrug system may require posology adjustment to produce wider beneficial effects on all muscle types.

Gait mechanics of lemurid primates on terrestrial and arboreal substrates

Aspects of gait mechanics of two lemurid species were explored experimentally. Substrate reaction forces were recorded for three animals each of L. catta and E. fulvus walking and running at voluntary speeds either on a wooden runway with an integrated force platform or on elevated pole supports with a section attached to the force platform. The average height of the back over these substrates and fluctuations in this height were evaluated using video-analysis. Animals preferred walking gaits and lower speeds on the poles, and gallops and higher speeds on the ground. At overlapping speeds, few adjustments to substrate types were identified. Hind limb peak forces are usually lower on the poles than on the ground, and the caudal back is closer to the substrate. This suggests that greater hind limb flexion and reduced limb stiffness occurred on the poles. The support phases for both limbs at higher speeds are slightly elongated on the poles. Forelimb peak forces are not lower, and the trajectory of the caudal back does not follow a smoother path, i.e., not all elements of a compliant gait are present on the simulated arboreal substrates. The horizontal, rigid poles, offered as substitutes for branchlike supports in the natural habitat, may not pose enough of a challenge to require more substantial gait adjustments. Across substrates, forelimb peak forces are generally lower than hind limb peak forces. The interlimb force distribution is similar to that of most other primates with more even limb lengths. Walking gaits present a greater divergence in fore- and hind limb forces than galloping gaits, which are associated with higher forces. The more arboreal E. fulvus has higher forelimb forces than the more terrestrial L. catta, unlike some anthropoid species in which the arborealists have lower forelimb forces than the terrestrialists. As in other primate and nonprimate quadrupeds, the major propulsive thrust comes from the hind limbs in both lemurs. While our data confirm certain aspects of primate gait mechanics (e.g., generally higher hind limb forces), they do not fully support the notion of greater limb compliance. Neither a compliant forelimb on branchlike supports, nor a negative correlation of forelimb force magnitudes with degree of arboreality were observed. Increasing forelimb-to-hind-limb-force-ratios with increasing speed and force magnitudes are also not expected under this paradigm.