Quantification of surface EMG signals to monitor the effect of a Botox treatment in six healthy ponies and two horses with stringhalt: preliminary study

Reasons for performing the study: Therapeutic options for stringhalt in horses are limited, whereas medical experiences with botulinum toxin type A (Botox) have been positive. To evaluate its effectiveness in horses, surface electromyography (sEMG) signals before and after injection need to be quantified. Hypothesis: Treatment of healthy ponies and cases with Botox should reduce muscle activity in injected muscles and reduce spastic movements without adverse side effects. Methods: Unilaterally, the extensor digitorum longus, extensor digitorum lateralis and lateral vastus muscles of 6 healthy mature Shetland ponies and 2 talented Dutch Warmblood dressage horses with stringhalt were injected (maximum of 400 iu per pony and 700 in per case; 100 in in 5 ml NaCl divided into 5 injections) with Botox under needle EMG guidance. Surface EMG data were evaluated using customised software, and in the individuals gait was analysed using Proreflex. Statistical analysis was performed using mixed models and independent sample t test (P<0.05). Results: Surface EMG signals were quantified using customised software. The area under the curve (integrated EMG) in time was used as variable. It became significantly reduced in injected muscles after injection of Botox in normal ponies (P<0.05). This effect was present from Day 1 until Day 84 after injection. In the 2 cases, after injection of 3 muscles, the integrated EMG in time became significantly reduced in all 3 muscles. Kinematic measurements confirmed reduction of frequency and amplitude of hyperflexing or hyperabducting strides of the affected hindlimbs. The duration of effect was also seen in the cases until around 12 weeks after injection. Conclusions and potential relevance: After EMG guided injections of Botox, sEMG signals recorded from injected muscle were reduced, which proves this to be a useful tool in statistically evaluating a treatment effect. The positive results of this pilot study encourage further research with a larger group of clinical cases

A pilot study on objective quantification and anatomical modelling of in vivo head and neck positions commonly applied in training and competition of sport horses

Reasons for performing study: Head and neck positions (HNP) in sport horses are under debate in the equine community, as they could interfere with equine welfare. HNPs have not been quantified objectively and no information is available on their head and neck loading. Objectives: To quantify in vivo HNPs in sport horses and develop o a model to estimate loading on the cervical vertebrae in these positions. Methods: Videos were taken of 7 Warmbloods at walk on a straight line in 5 positions, representing all HNPs during Warmblood training and competition. Markers were glued at 5 anatomical landmarks. Two-dimensional angles and distances were determined from video frames for the 5 HNPs and statistically compared (P < 0.05). A new simulation model was developed to estimate nuchal ligament cervical loading at these HNPs. Results: The mean angles were significantly different between the 5 HNPs for the line between C1 and T6 with the horizontal and for the line connecting the facial crest (CF) and C1 with the vertical, while the vertical distance from CF to the lateral styloid process of the radius (PS) was significantly different between all 5 positions (P < 0.05). The estimated nuchal ligament loading appeared to be largest at the origin of C2 for all HNPs, except for the 'hyperextended' HNP5; the 'hyperflexed' HNP4 showed the largest loading values on the nuchal ligament origins at all locations. Conclusions: HNPs can be accurately quantified in the sagittal plane from angles and distances based on standard anatomical landmarks and home-video captured images. Nuchal ligament loading showed the largest estimated values at its origin on C2 in hyperflexion (HNP4). Potential relevance: Modelling opens further perspectives to eventually estimate loading for individual horses and thus ergonomically optimise their HNP, which may improve the welfare of the sport horse during training and competition

Expanding Clinical Presentations Due to Variations in THOC2 mRNA Nuclear Export Factor.

Multiple TREX mRNA export complex subunits (e.g., THOC1, THOC2, THOC5, THOC6, THOC7) have now been implicated in neurodevelopmental disorders (NDDs), neurodegeneration and cancer. We previously implicated missense and splicing-defective THOC2 variants in NDDs and a broad range of other clinical features. Here we report 10 individuals from nine families with rare missense THOC2 variants including the first case of a recurrent variant (p.Arg77Cys), and an additional individual with an intragenic THOC2 microdeletion (Del-Ex37-38). Ex vivo missense variant testing and patient-derived cell line data from current and published studies show 9 of the 14 missense THOC2 variants result in reduced protein stability. The splicing-defective and deletion variants result in a loss of small regions of the C-terminal THOC2 RNA binding domain (RBD). Interestingly, reduced stability of THOC2 variant proteins has a flow-on effect on the stability of the multi-protein TREX complex; specifically on the other NDD-associated THOC subunits. Our current, expanded cohort refines the core phenotype of THOC2 NDDs to language disorder and/or ID, with a variable severity, and disorders of growth. A subset of affected individuals' has severe-profound ID, persistent hypotonia and respiratory abnormalities. Further investigations to elucidate the pathophysiological basis for this severe phenotype are warranted

αCaMKII Autophosphorylation Controls the Establishment of Alcohol Drinking Behavior

The α-Ca(2+)/calmodulin-dependent protein kinase II (αCaMKII) is a crucial enzyme controlling plasticity in the brain. The autophosphorylation of αCaMKII works as a ‘molecular memory' for a transient calcium activation, thereby accelerating learning. We investigated the role of αCaMKII autophosphorylation in the establishment of alcohol drinking as an addiction-related behavior in mice. We found that alcohol drinking was initially diminished in αCaMKII autophosphorylation-deficient αCaMKII(T286A) mice, but could be established at wild-type level after repeated withdrawals. The locomotor activating effects of a low-dose alcohol (2 g/kg) were absent in αCaMKII(T286A) mice, whereas the sedating effects of high-dose (3.5 g/kg) were preserved after acute and subchronic administration. The in vivo microdialysis revealed that αCaMKII(T286A) mice showed no dopamine (DA) response in the nucleus accumbens to acute or subchronic alcohol administration, but enhanced serotonin (5-HT) responses in the prefrontal cortex. The attenuated DA response in αCaMKII(T286A) mice was in line with altered c-Fos activation in the ventral tegmental area after acute and subchronic alcohol administration. In order to compare findings in mice with the human condition, we tested 23 single-nucleotide polymorphisms (SNPs) in the CAMK2A gene for their association with alcohol dependence in a population of 1333 male patients with severe alcohol dependence and 939 controls. We found seven significant associations between CAMK2A SNPs and alcohol dependence, one of which in an autophosphorylation-related area of the gene. Together, our data suggest αCaMKII autophosphorylation as a facilitating mechanism in the establishment of alcohol drinking behavior with changing the DA–5-HT balance as a putative mechanism