Generation of Spike-Extracellular Vesicles (S-EVs) as a Tool to Mimic SARS-CoV-2 Interaction with Host Cells

Extracellular vesicles (EVs) and viruses share common features: size, structure, biogenesis and uptake. In order to generate EVs expressing the SARS-CoV-2 spike protein on their surface (S-EVs), we collected EVs from SARS-CoV-2 spike expressing human embryonic kidney (HEK-293T) cells by stable transfection with a vector coding for the S1 and S2 subunits. S-EVs were characterized using nanoparticle tracking analysis, ExoView and super-resolution microscopy. We obtained a population of EVs of 50 to 200 nm in size. Spike expressing EVs represented around 40% of the total EV population and co-expressed spike protein with tetraspanins on the surfaces of EVs. We subsequently used ACE2-positive endothelial and bronchial epithelial cells for assessing the internalization of labeled S-EVs using a cytofluorimetric analysis. Internalization of S-EVs was higher than that of control EVs from non-transfected cells. Moreover, S-EV uptake was significantly decreased by anti-ACE2 antibody pre-treatment. Furthermore, colchicine, a drug currently used in clinical trials, significantly reduced S-EV entry into the cells. S-EVs represent a simple, safe, and scalable model to study host-virus interactions and the mechanisms of novel therapeutic drugs.

Lower extremity kinematic coupling during single and double leg landing and gait in female junior athletes with dynamic knee valgus

Background Dynamic knee valgus (DKV) is a common lower extremity movement disorder among females. This study aimed to investigate kinematic couplings between lower extremity joints in female junior athletes with DKV during single and double-leg landing and gait. Methods Twenty-six physically active female junior athletes (10–14 years old) with DKV were recruited. Kinematic couplings between rearfoot, tibia, knee, and hip were extracted using eight Vicon motion capture cameras and two force plates. Zero-lag cross-correlation coefficient and vector coding were used to calculate kinematic couplings between joints during physical tasks. Paired t-test and Wilcoxon tests were run to find significant couplings between joint motions and coupling strengths. Bonferroni posthoc was used to determine significance with α ≤ 0.05. Results The results showed that the strongest kinematic relationship existed between rearfoot eversion/inversion and tibial internal/external rotation during all three tasks. Correlations of the rearfoot supination/pronation with tibial rotations, knee, and hip motions in sagittal, frontal, and transverse planes were very strong to strong during double-leg landing and moderate to weak during gait. A weak correlation was observed between rearfoot supination/pronation and hip adduction/abduction during single-leg landing. Conclusions Coupling relationships between rearfoot, knee, and hip vary by the task intensity and alignment profiles in female juniors with DKV.

Advancements in data analysis and visualisation techniques to support multiple single-subject analyses: an assessment of movement coordination and coordination variability

Vector coding is a data analysis technique that quantifies inter-segmental coordination and coordination variability of human movement. The usual reporting of vector coding time-series data can be difficult to interpret when multiple trials are superimposed on the same figure. This study describes and presents novel data visualisations for displaying data from vector coding that supports multiple single- subject analyses. The dataset used in this study describes the lumbar-pelvis coordination in the transverse plane during a gait cycle. The data visualisation techniques presented in this study consists of the use of colour and data bars to map and profile coordination pattern and coordination variability data. The use of colour mapping provides the option to classify commonalities and differences in patterns of coordination between segment couplings and between individuals across a big dataset. Data bars display segmental dominancy data that can provide an intuitive summary on coupling angle distribution over time. The data visualisation in this study may provide further insight on how people with adolescent idiopathic scoliosis perform goal-orientated movements following an intervention, which would support clinical management strategies.

Glucocerebrosidase Gene Therapy Induces Alpha-Synuclein Clearance and Neuroprotection of Midbrain Dopaminergic Neurons in Mice and Macaques

Mutations in the GBA1 gene coding for glucocerebrosidase (GCase) are the main genetic risk factor for Parkinson’s disease (PD). Indeed, identifying reduced GCase activity as a common feature underlying the typical neuropathological signatures of PD—even when considering idiopathic forms of PD—has recently paved the way for designing novel strategies focused on enhancing GCase activity to reduce alpha-synuclein burden and preventing dopaminergic cell death. Here we have performed bilateral injections of a viral vector coding for the mutated form of alpha-synuclein (rAAV9-SynA53T) for disease modeling purposes, both in mice as well as in nonhuman primates (NHPs), further inducing a progressive neuronal death in the substantia nigra pars compacta (SNpc). Next, another vector coding for the GBA1 gene (rAAV9-GBA1) was unilaterally delivered in the SNpc of mice and NHPs one month after the initial insult, together with the contralateral delivery of an empty/null rAAV9 for control purposes. Obtained results showed that GCase enhancement reduced alpha-synuclein burden, leading to improved survival of dopaminergic neurons. Data reported here support using GCase gene therapy as a disease-modifying treatment for PD and related synucleinopathies, including idiopathic forms of these disorders.