RESUMEN
Exosomes are nanosized extracellular vesicles secreted by all cell types, including canine adipose-derived stem cells (cADSCs). By mediating intercellular communication, exosomes modulate the biology of adjacent and distant cells by transferring their cargo. In the present work after isolation and characterization of exosomes derived from canine adipose tissue, we treated the same canine donors affected by hepatopathies with the previously isolated exosomes. We hypothesize that cADSC-sourced miRNAs are among the factors responsible for a regenerative and anti-inflammatory effect in the treatment of hepatopathies in dogs, providing the clinical veterinary field with an effective and innovative cell-free therapy. Exosomes were isolated and characterized for size, distribution, surface markers, and for their miRNomic cargo by microRNA sequencing. 295 dogs affected with hepatopathies were treated and followed up for 6 months to keep track of their biochemical marker levels. Results confirmed that exosomes derived from cADSCs exhibited an average diameter of 91 nm, and positivity to 8 known exosome markers. The administration of exosomes to dogs affected by liver-associated inflammatory pathologies resulted in the recovery of the animal alongside the normalization of biochemical parameters of kidney function. In conclusion, cADSCs-derived exosomes are a promising therapeutic tool for treating inflammatory disorders in animal companions.
Asunto(s)
Exosomas , Vesículas Extracelulares , MicroARNs , Perros , Animales , MicroARNs/genética , Exosomas/genética , Exosomas/metabolismo , Vesículas Extracelulares/metabolismo , Hepatitis Crónica/metabolismo , Células Madre/metabolismoRESUMEN
Monoamine transporters are of great interest for their role in the physiological activity of the body and their link to mental and behavioural disorders. Currently, static well-plate assays or manual perfusion systems are used to characterize the interaction of psychostimulants, antidepressants and drugs of abuse with the transporters but still suffer from significant drawbacks caused by lack of automation, for example, low reproducibility, non-comparability of results. An automated microfluidic platform was developed to address the need for more standardized procedures for cell-based assays. An automated system was used to control and drive the simultaneous perfusion of 12 channels on a microfluidic chip, establishing a more standardized protocol to perform release assays to study monoamine transporter-mediated substrate efflux. D-Amphetamine, GBR12909 (norepinephrine transporter) and p-chloroamphetamine, paroxetine (serotonin transporter) were used as control compounds to validate the system. The platform was able to produce the expected releasing (D-Amphetamine, p-chloroamphetamine) or inhibiting (GBR12909, paroxetine) profiles for the two transporters. The reduction of manual operation and introduction of automated flow control enabled the implementation of stronger standardized protocols and the possibility of obtaining higher throughput by increasing parallelization.
Asunto(s)
Microfluídica , p-Cloroanfetamina , Paroxetina , Reproducibilidad de los Resultados , Proteínas de Transporte de Membrana , Perfusión , DextroanfetaminaRESUMEN
The epidemiological characteristics of spontaneous hypercortisolism (HC) were derived from 21,281 client-owned dogs selected from four private veterinary clinics and one university reference center for endocrinology. The odds ratio (OR) method was employed to investigate the risk of developing HC related to breed, gender, and sexual status. The estimated prevalence of HC in the four private clinics was 0.20% [95% confidence interval (CI), 0.13-0.27] and was significantly different compared to the university reference center (1.46%; 95% CI, 1.12-1.80). Sex, breed, and age resulted in risk factors for HC. Mean (± SD) age for dogs with HC was 9.8 (± 2.5) yr. Females had higher risk for HC compared to males (OR 1.85; 95% CI, 1.24-2.75); all neutered dogs (both males and females) had higher risk than intact dogs (OR 2.54; 95% CI, 1.72-3.73); and neutered females had higher risk compared to intact females (OR 2.61; 95% CI, 1.54-4.42). Using the mixed breed dogs as a control population (OR = 1), the risk of developing HC was significantly higher in the Standard Schnauzer (OR 58.1; p < 0.0001) and Fox Terrier (OR 20.33; p < 0.0001). With regard to HC, this study identified an overall prevalence of 0.20%. The data support the existence of sex predisposition, with the highest risk for neutered females.