Large-scale bioreactor production of extracellular vesicles from mesenchymal stromal cells for treatment of acute radiation syndrome.

BACKGROUND: Hematopoietic acute radiation syndrome (H-ARS) occurring after exposure to ionizing radiation damages bone marrow causing cytopenias, increasing susceptibility to infections and death. We and others have shown that cellular therapies like human mesenchymal stromal cells (MSCs), or monocytes/macrophages educated ex-vivo with extracellular vesicles (EVs) from MSCs were effective in a lethal H-ARS mouse model. However, given the complexity of generating cellular therapies and the potential risks of using allogeneic products, development of an "off-the-shelf" cell-free alternative like EVs may have utility in conditions like H-ARS that require rapid deployment of available therapeutics. The purpose of this study was to determine the feasibility of producing MSC-derived EVs at large scale using a bioreactor and assess critical quality control attributes like identity, sterility, and potency in educating monocytes and promoting survival in a lethal H-ARS mouse model. METHODS: EVs were isolated by ultracentrifugation from unprimed and lipopolysaccharide (LPS)-primed MSCs grown at large scale using a hollow fiber bioreactor and compared to a small scale system using flasks. The physical identity of EVs included a time course assessment of particle diameter, yield, protein content and surface marker profile by flow-cytometry. Comparison of the RNA cargo in EVs was determined by RNA-seq. Capacity of EVs to generate exosome educated monocytes (EEMos) was determined by qPCR and flow cytometry, and potency was assessed in vivo using a lethal ARS model with NSG mice. RESULTS: Physical identity of EVs at both scales were similar but yields by volume were up to 38-fold more using a large-scale bioreactor system. RNA-seq indicated that flask EVs showed upregulated let-7 family and miR-143 micro-RNAs. EEMos educated with LPS-EVs at each scale were similar, showing increased gene expression of IL-6, IDO, FGF-2, IL-7, IL-10, and IL-15 and immunophenotyping consistent with a PD-L1 high, CD16 low, and CD86 low cell surface expression. Treatment with LPS-EVs manufactured at both scales were effective in the ARS model, improving survival and clinical scores through improved hematopoietic recovery. EVs from unprimed MSCs were less effective than LPS-EVs, with flask EVs providing some improved survival while bioreactor EVs provide no survival benefit. CONCLUSIONS: LPS-EVs as an effective treatment for H-ARS can be produced using a scale-up development manufacturing process, representing an attractive off-the-shelf, cell-free therapy.

Stress response gene family expansions correlate with invasive potential in teleost fish.

The bluegill sunfish Lepomis macrochirus and the closely related redear sunfish Lepomis microlophus have important ecological and recreational value and are widely used for research and aquaculture. While both species have been introduced outside of their native ranges, only the bluegill is considered invasive. Here, we report de novo transcriptome assemblies for these fish as a resource for sunfish biology. Comparative analyses of the transcriptomes revealed an unexpected, bluegill-specific expansion in the HSP70 and HSP90 molecular chaperone gene families. These expansions were not unique to the bluegill as expansions in HSP70s and HSP90s were identified in the genomes of other teleost fish using the NCBI RefSeq database. To determine whether gene family expansions are specific for thermal stress responses, GST and SOD gene families that are associated with oxidative stress responses were also analyzed. Species-specific expansions were also observed for these gene families in distinct fish species. Validating our approach, previously described expansions in the MHC gene family were also identified. Intriguingly, the number of HSP70 paralogs was positively correlated with thermotolerance range for each species, suggesting that these expansions can impact organismal physiology. Furthermore, fish that are considered invasive contained a higher average number of HSP70 paralogs than non-invasive fish. Invasive fish also had higher average numbers of HSP90, MHC and GST paralogs, but not SOD paralogs. Taken together, we propose that expansions in key cellular stress response gene families represent novel genetic signatures that correlate with invasive potential.

Identification of sample mix-ups and mixtures in microbiome data in Diversity Outbred mice.

In a Diversity Outbred mouse project with genotype data on 500 mice, including 297 with microbiome data, we identified three sets of sample mix-ups (two pairs and one trio) as well as at least 15 microbiome samples that appear to be mixtures of pairs of mice. The microbiome data consisted of shotgun sequencing reads from fecal DNA, used to characterize the gut microbial communities present in these mice. These sequence reads included sufficient reads derived from the host mouse to identify the individual. A number of microbiome samples appeared to contain a mixture of DNA from two mice. We describe a method for identifying sample mix-ups in such microbiome data, as well as a method for evaluating sample mixtures in this context.

Efeitos de um treino em ambiente virtual sobre o desempenho da marcha e funções cognitivas em idosos saudáveis / Effects of a training in virtual environment about gait performance and cognitive functions in healthy elderly

O objetivo do presente estudo foi comparar os efeitos de dois tipos de programas de treinamento do equilíbrio e marcha, um baseado no Nintendo Wii Fit® e o outro baseado em exercícios convencionais sem a utilização do videogame, na eficiência da marcha, funções cognitivas e funcionalidade de idosos saudáveis. Trata-se de um ensaio clínico, cego e randomizado realizado no Departamento de Fonoaudiologia, Fisioterapia e Terapia Ocupacional no Laboratório de Aprendizagem Sensório-Motora (LASM) da Universidade de São Paulo. Participaram do estudo 32 idosos saudáveis que foram randomizados em grupo experimental (n=16) e grupo controle (n=16). Ambos os grupos realizaram 14 sessões individuais de treinamento, duas vezes na semana por sete semanas. As sessões eram divididas em duas partes: a primeira parte era composta de 30 minutos de exercícios globais que incluíam alongamento e fortalecimento muscular além de mobilidade axial; a segunda parte era composta de 30 minutos de exercícios de equilíbrio e marcha. O grupo experimental realizou os exercícios de equilíbrio e marcha por meio de 10 jogos do vídeogame Nintendo Wii Fit®, que promoviam a estimulação cognitiva e motora por meio da realimentação visual e auditiva. Já o grupo controle realizou os exercícios sem nenhuma estimulação cognitiva associada. As principais medidas do estudo foram: (1) Functional Gait Assessment (FGA); (2) Montreal Cognitive Assessment (MoCA); (3) Índice do Andar Dinâmico (IAD); (4) Falls Efficacy Scale (FES-I); (5) Teste da marcha de 30 segundos em condição de simples tarefa e dupla tarefa. A análise estatística foi realizada por meio da ANOVA de medidas repetidas e pós hoc teste de Tukey para a verificação de possíveis diferenças entre grupos e avaliações, que foram realizadas antes, depois e após 60 dias do término do treinamento(AU)

The aim of this study was to compare the effects of two types of balance and gait training programs, one based on Nintendo Wii Fit® and the other based on conventional exercises without the use of video games, in the gait efficiency, cognitive functions and functionality in healthy elderly. This is a blind and randomized clinical trial, conducted at the Department of Speech-Language Pathology, Physiotherapy and Occupational Therapy at the Laboratory for Sensory-Motor Learning (LASM) at the University of São Paulo. The study included 32 healthy elderly individuals who were randomized into an experimental group (n = 16) and control group (n = 16). Both groups underwent 14 sessions of individual training, twice a week for seven weeks. The sessions were divided into two parts: the first part consisted of 30-minute global exercise that included stretching and muscular strengthening exercises as well as axial mobility; the second part consisted of 30 minutes of balance and gait exercises. The experimental group performed balance and gait exercises through 10 Nintendo Wii Fit® video games, which promoted cognitive and motor stimulation through visual and auditory feedback. The control group performed the exercises without any cognitive stimulation associated. The main measures of the study were: (1) Functional Gait Assessment (FGA), (2) Montreal Cognitive Assessment (MoCA), (3) Dynamic Gait Index (DGI), (4) Falls Efficacy Scale (FES-I); (5) 30-second gait test in simple and dual-task condition. Statistical analysis was performed by repeated measures ANOVA and post hoc Tukey test to check for possible differences among groups and evaluations, which were performed before, after and 60 days after the end of the training(AU)

Effect of Nintendo Wii™-based motor and cognitive training on activities of daily living in patients with Parkinson's disease: a randomised clinical trial.

OBJECTIVES: To investigate the effect of Nintendo Wii™-based motor cognitive training versus balance exercise therapy on activities of daily living in patients with Parkinson's disease. DESIGN: Parallel, prospective, single-blind, randomised clinical trial. SETTING: Brazilian Parkinson Association. PARTICIPANTS: Thirty-two patients with Parkinson's disease (Hoehn and Yahr stages 1 and 2). INTERVENTIONS: Fourteen training sessions consisting of 30 minutes of stretching, strengthening and axial mobility exercises, plus 30 minutes of balance training. The control group performed balance exercises without feedback or cognitive stimulation, and the experimental group performed 10 Wii Fit™ games. MAIN OUTCOME MEASURE: Section II of the Unified Parkinson's Disease Rating Scale (UPDRS-II). RANDOMISATION: Participants were randomised into a control group (n=16) and an experimental group (n=16) through blinded drawing of names. STATISTICAL ANALYSIS: Repeated-measures analysis of variance (RM-ANOVA). RESULTS: Both groups showed improvement in the UPDRS-II with assessment effect (RM-ANOVA P<0.001, observed power=0.999). There was no difference between the control group and the experimental group before training {8.9 [standard deviation (SD) 2.9] vs 10.1 (SD 3.8)}, after training [7.6 (SD 2.9) vs 8.1 (SD 3.5)] or 60 days after training [8.1 (SD 3.2) vs 8.3 (SD 3.6)]. The mean difference of the whole group between before training and after training was -0.9 (SD 2.3, 95% confidence interval -1.7 to -0.6). CONCLUSION: Patients with Parkinson's disease showed improved performance in activities of daily living after 14 sessions of balance training, with no additional advantages associated with the Wii-based motor and cognitive training. Registered on http://www.clinicaltrials.gov (identifier: NCT01580787).

Motor learning, retention and transfer after virtual-reality-based training in Parkinson's disease--effect of motor and cognitive demands of games: a longitudinal, controlled clinical study.

OBJECTIVES: To evaluate the learning, retention and transfer of performance improvements after Nintendo Wii Fit™ training in patients with Parkinson's disease and healthy elderly people. DESIGN: Longitudinal, controlled clinical study. PARTICIPANTS: Sixteen patients with early-stage Parkinson's disease and 11 healthy elderly people. INTERVENTIONS: Warm-up exercises and Wii Fit training that involved training motor (shifts centre of gravity and step alternation) and cognitive skills. A follow-up evaluative Wii Fit session was held 60 days after the end of training. Participants performed a functional reach test before and after training as a measure of learning transfer. MAIN OUTCOME MEASURES: Learning and retention were determined based on the scores of 10 Wii Fit games over eight sessions. Transfer of learning was assessed after training using the functional reach test. RESULTS: Patients with Parkinson's disease showed no deficit in learning or retention on seven of the 10 games, despite showing poorer performance on five games compared with the healthy elderly group. Patients with Parkinson's disease showed marked learning deficits on three other games, independent of poorer initial performance. This deficit appears to be associated with cognitive demands of the games which require decision-making, response inhibition, divided attention and working memory. Finally, patients with Parkinson's disease were able to transfer motor ability trained on the games to a similar untrained task. CONCLUSIONS: The ability of patients with Parkinson's disease to learn, retain and transfer performance improvements after training on the Nintendo Wii Fit depends largely on the demands, particularly cognitive demands, of the games involved, reiterating the importance of game selection for rehabilitation purposes.