AAV8 vector induced gliosis following neuronal transgene expression.

Introduction: Expression of light sensitive ion channels by selected neurons has been achieved by viral mediated transduction with gene constructs, but for this to have therapeutic uses, for instance in treating epilepsy, any adverse effects of viral infection on the cerebral cortex needs to be evaluated. Here, we assessed the impact of adeno-associated virus 8 (AAV8) carrying DNA code for a soma targeting light activated chloride channel/FusionRed (FR) construct under the CKIIa promoter. Methods: Viral constructs were harvested from transfected HEK293 cells in vitro and purified. To test functionality of the opsin, cultured rodent neurons were transduced and the light response of transduced neurons was assayed using whole-cell patch-clamp recordings. In vivo expression was confirmed by immunofluorescence for FR. Unilateral intracranial injections of the viral construct were made into the mouse neocortex and non-invasive fluorescence imaging of FR expression made over 1-4 weeks post-injection using an IVIS Spectrum system. Sections were also prepared from injected mouse cortex for immunofluorescence staining of FR, alongside glial and neuronal marker proteins. Results: In vitro, cortical neurons were successfully transduced, showing appropriate physiological responses to light stimulation. Following injections in vivo, transduction was progressively established around a focal injection site over a 4-week period with spread of transduction proportional to the concentration of virus introduced. Elevated GFAP immunoreactivity, a marker for reactive astrocytes, was detected near injection sites associated with, and proportional to, local FR expression. Similarly, we observed reactive microglia around FR expressing cells. However, we found that the numbers of NeuN+ neurons were conserved close to the injection site, indicating that there was little or no neuronal loss. In control mice, injected with saline only, astrocytosis and microgliosis was limited to the immediate vicinity of the injection site. Injections of opsin negative viral constructs resulted in comparable levels of astrocytic reaction as seen with opsin positive constructs. Discussion: We conclude that introduction of an AAV8 vector transducing expression of a transgene under a neuron specific promotor evokes a mild inflammatory reaction in cortical tissue without causing extensive short-term neuronal loss. The expression of an opsin in addition to a fluorescent protein does not significantly increase neuroinflammation.

Injuries outside of horseracing: is it time to focus on injury prevention of jockeys outside of races?

Objectives: Falls and injuries are frequent in professional horseracing. However, professional jockeys spend a large part of their week in horse-related activities outside of racing such as schooling, exercise riding, and yard-related activities. The injury risk related to these activities remains largely unknown internationally. This study aimed to identify the injury prevalence and injury profile of flat and jump jockeys during non-racing activities. Methods: In total 45.6% and 38.5% of all licensed Irish professional flat and jump jockeys completed a cross-sectional self-recall questionnaire examining injuries that occurred outside of racing during 2018. Injury proportion, repeat incidence proportion, and descriptive statistics were calculated. Results: Fifteen percent of professional jockeys sustained an injury outside of a race, half of those injured receiving at least another injury in 2018 and 66.52 injuries per 1,000 falls were noted. Injuries frequently occurred to the upper limb (36.67%), with fractures common (32.00%). Most injuries occurred due to a fall (60.00%) and 77.27% occurred in the gallops. Half of injuries resulted in the jockey missing racing, with 31.00 ± 47.18 (4-180) days lost on average. Twenty-three percent of jockeys attended hospital and 16.67% required surgery due to injury. Interestingly, just under a third did not report their injury to anyone. Conclusion: Injuries to professional jockeys, whilst not as frequent outside of racing, tend to be serious and can affect jockeys financially and impact their availability to ride. Prioritizing injury prevention strategies to maximize availability of jockeys to race is important. Education on the importance of reporting all injuries regardless of where they occur is important to ensure their management and rehabilitation.

Injuries in Collegiate Ladies Gaelic Footballers: A 2-Season Prospective Cohort Study.

CONTEXT: Although Ladies Gaelic football is one of the most popular female sports in Ireland, just 2 previous injury surveillance studies have been completed, and both were retrospective in nature. OBJECTIVE: To prospectively examine the injury incidence and injury profile in collegiate Ladies Gaelic football over 2 seasons. DESIGN: Prospective cohort study. SETTING: College. Patients (or Other Participants): Adult Ladies Gaelic footballers from one collegiate institution (season 1: n = 50, season 2: n = 82). INTERVENTION(S): All time-loss injuries that occurred were recorded by certified athletic therapists and student-athletic therapists and trainers over 2 seasons. MAIN OUTCOME MEASURES: A standardized injury report form was used to record the injury onset, mechanism, location, nature, and outcome. Injury incidence proportion, repeat incidence proportion and total, match and training injury rates, and their 95% confidence intervals were calculated. The frequencies and proportions were also calculated. RESULTS: The match and training injury rates were 42.48 and 7.93 injuries per 1000 hours, respectively. A low repeat incidence proportion per season was noted (11.7% and 0.0%). The injuries were predominantly acute (74.68%) and noncontact (66.25%), with hamstring injuries (21.52%) and strains (36.71%) the most frequent location and nature of injuries noted. Strains (104.92 d absent per 1000 h) and knee injuries (106.46 d absent per 1000 h) led to the greatest injury burden. Further investigations were not frequently required, with an X-ray and magnetic resonance imaging ordered in 8.00% and 6.67% of the cases, respectively. Surgery was completed following one injury. CONCLUSIONS: This is the first study to provide prospective injury data on Ladies Gaelic football. Priority needs to be given to preventing hamstring and knee injuries due to their occurrence and negative impact on player availability to play. Collegiate Ladies Gaelic football teams should be encouraged to implement an injury-prevention warm-up, such as the GAA15+, at training and matches.