Extracellular vesicles from equine mesenchymal stem cells decrease inflammation markers in chondrocytes in vitro

Background: Mesenchymal stem cells (MSCs) have been used therapeutically in equine medicine. MSCs release extracellular vesicles (EVs), which affect cell processes by inhibiting cell apoptosis and regulating inflammation. To date, little is known about equine EVs and their regenerative properties. Objectives: To characterise equine MSC-derived extracellular vesicles (EVs) and evaluate their effect on equine chondrocytes treated with pro-inflammatory cytokines in vitro. Study design: In vitro experiments with randomised complete block design.Methods: Mesenchymal stem cells from bone marrow, adipose tissue, and synovial fluid were cultured in vitro. The MSC culture medium was centrifuged and filtered. Isolated particles were analysed for size and concentration (total number of particles per mL). Transmission electron microscopy analysis was performed to evaluate the morphology and CD9 expression of the particles. Chondrocytes from healthy equines were treated with the inflammatory cytokines interleukin (IL)-1β and tumour necrosis factor-alpha. MSC-derived EVs from bone marrow and synovial fluid cells were added as co-treatments in vitro. Gene expression analysis by real-time PCR was performed to evaluate the effects of EVs. Results: The particles isolated from MSCs derived from different tissues did not differ significantly in size and concentration. The particles had a round-like shape and positively expressed CD9. EVs from bone marrow cells displayed reduced expression of metalloproteinase-13. Main limitations: Sample size and characterisation of the content of EVs Conclusions: EVs isolated from equine bone marrow MSCs reduced metalloproteinase 13 gene expression; this gene encodes an enzyme related to cartilage degradation in inflamed chondrocytes in vitro. EVs derived from MSCs can reduce inflammation and could potentially be used as an adjuvant treatment to improve tissue and cartilage repair in the articular pathologies

A randomised controlled trial of a school-based intervention to prevent tobacco use among children and adolescents in Italy

Aim: A randomised field trial was conducted to evaluate a school-based programme to prevent tobacco use in children and adolescents. Subject and methods: The trial included 534 children and 308 adolescents who were randomly selected to receive or not to receive the prevention programme. The prevention programme included: (a) health facts and the effect of smoking, (b) analysis of the mechanisms underlying intiation of smoking and (c) refusal skills training to deal with the social pressures to smoke. A questionnaire was administered before the intervention programme and 2 years later. Results: The prevalence rates of smoking in both groups of children and adolescents were increased at the end of the study. Anyway, the difference of smoking prevalence between the intervention and control groups was statistically significant only for the children’s group (from 18.3 to 18.8% for the intervention group and from 17.8 to 26.9% in the control group) (p = 0.035). As regards reasons that induced the start of smoking, there was a significant increase of the issue "because smokers are fools" (p = 0.004 for children; p < 0.001 for adolescents) and "because smokers are irresponsible" (p ≤ 0.001 for both children and adolescents) in the experimental groups. Conclusion: The results suggest that a school-based intervention programme addressing tobacco use among children and adolescents, based on the development of cognitive and behavioural aspects, can be effective. After 1 year of intervention, smoking prevalence was significantly lower in children belonging to the intervention group than in children not randomised to intervention. Targeting young children before they begin to smoke can be a successful way of prevention

Knowledge, attitudes and behaviour about Sexually Transmitted Infections: a survey among Italian university female students

Aims: Improvements have been made in terms of STIs mortality, but young women still have a lack of attention to the STIs prevention. We investigated the knowledge and attitudes about STIs and their prevention among a sample of Italian university female students. Methods: We designed a questionnaire about knowledge and attitudes toward STIs, Papanicolau test (Pap test) and Human Papilloma Virus (HPV) and had it filled out by 285 young women. Chi-square test, Mann-Whitney test and a multivariable analysis were performed in order to identify associated factors. &nbsp; Results: Knowledge about STIs seems weak, oppositely to knowledge about preventive measures. In line with the results of the univariate analyses, the multivariable one confirmed a higher probability to answer rightly for women who knew the effective behaviours for preventing STIs, women who took information about STIs from Internet, books and TV and those using natural methods (OR 2.77 - 95%CI 1.47-5.23, OR 2.99 - 95%CI 1.62-5.53 and OR 2.55 - 95%CI 1.05-6.18, respectively) and a lower one for students from Cassino and those who knew is possible to recover from STIs (OR 0.38, 95%CI 0.17-0.87 and 0.42, 95%CI 0.20-0.88, respectively). Furthermore, about Pap test it also confirmed the role of a medium-high STIs level of knowledge (OR 2.11, 95%CI 1.02-4.37) and a lower probability to know about it for women from Cassino (OR 0.25, 95%CI 0.09-0.73). Conclusions: Young women are not completely aware of STIs risks and preventive measures. Informational and educational campaigns should be implemented to reach this target group and lighten the current and future burden of STIs

Goat milk extracellular vesicles: immuno-modulation effects on porcine monocyte-derived macrophages in vitro

IntroductionExtracellular vesicles (EVs) are nanometric-membrane-bound sub-cellular structures, which can be recovered from milk. Milk EVs have drawn increasing interest due to their potential biomedical applications, therefore it is important to investigate their impact on key immune cells, such as macrophages.MethodsIn this work, the immunomodulatory effects of goat milk EVs on untreated (moMФ) and classically activated (moM1) porcine monocyte-derived macrophages were investigated using flow cytometry, ELISA, and gene expression assays.ResultsThese particles were efficiently internalized by macrophages and high doses (60 mg protein weight) triggered the upregulation of MHC I and MHC II DR on moMФ, but not on moM1. In moMФ, exposure to low doses (0.6 mg) of mEVs enhanced the gene expression of IL10, EBI3, and IFNB, whereas high doses up-regulated several pro-inflammatory cytokines. These nanosized structures slightly modulated cytokine gene expression on moM1. Accordingly, the cytokine (protein) contents in culture supernatants of moMФ were mildly affected by exposure to low doses of mEVs, whereas high doses promoted the increased release of TNF, IL-8, IL-1a, IL-1b, IL-1Ra, IL-6, IL-10, and IL-12. The cytokines content in moM1 supernatants was not critically affected.DiscussionOverall, our data support a clinical application of these molecules: they polarized macrophages toward an M1-like phenotype, but this activation seemed to be controlled, to prevent potentially pathological over-reaction to stressors

Pharmacokinetics of cannabidiol following single oral and oral transmucosal administration in dogs

IntroductionIn the last few years, different formulations containing cannabidiol (CBD) were tested with regard to its efficacy on chronic pain, refractory epilepsy, anxiety, aggressive behavior and atopic dermatitis in dogs. CBD is generally administered orally, but its low bioavailability, probably due to a first-pass metabolism, represents a great limitation. The aim of this study was to evaluate if CBD bioavailability increases after oral transmucosal administration (OTM) compared to oral treatment.MethodsTwelve dogs diagnosed with mild chronic pain were enrolled in the study and treated once orally or OTM (6 dogs/group) with a pure CBD in oil formulation at a dosing rate of 1 mg/kg b.w. At prefixed time points, blood samples were collected to define CBD plasma concentrations vs. time profiles, and the main pharmacokinetics parameters were obtained by non-compartmental model.ResultsCBD Cmax, Tmax, terminal half-life and AUC0 − t were 206.77 ± 167 and 200.33 ± 158.33 ng/mL, 2.17 ± 0.98 and 1.92 ± 1.11 h, 2.67 ± 0.53 and 2.62 ± 0.64 h, 647.51 ± 453.17, and 536.05 ± 370.21 h*ng/mL, following oral and OTM administration, respectively. No significant difference in pharmacokinetic parameters were observed between treatments.DiscussionThe OTM administration did not increase cannabidiol bioavailability compared to oral treatment. The almost perfectly superimposable mean plasma concentrations of cannabidiol following the two treatments suggests that CBD is not able to be adsorbed by the oral mucosa or that its absorption is very scarce, and that CBD is swallowed and absorbed in the gastrointestinal tract

Socio-demographic determinants of coinfections by HIV, hepatitis B and hepatitis C viruses in central Italian prisoners

BACKGROUND: The coinfections HIV/HCV/HBV are an important health issue in penitentiary communities. The aim of the study was to examine HIV, HBV and HCV coinfections determinants amongst prisoners in the jails of Southern Lazio (Central Italy), in the period 1995-2000. METHODS: Diagnosis of seropositivities for HIV, HBV and HCV was made using ELISA method. A multiple logistic regression analysis was conducted to verify the influence of socio-demographic factors on the HIV/HBV/HCV coinfections. RESULTS: HIV/HCV, HBV/HCV and HIV/HBV coinfections were detected in 42 (4%), 203 (17.9%) and 31 (2.9%) inmates, respectively. These coinfections are significantly associated with the status of drug addiction (OR = 16.02; p = 0.012; OR = 4.15; p &lt; 0.001; OR = 23.57; p = 0.002), smoking habits (OR = 3.73; p = 0.033; OR = 1.42; p = 0.088; OR = 4.25; p = 0.053) and Italian nationality (OR = 7.05; p = 0.009; OR = 2.31; p &lt; 0.001; OR = 4.61; p = 0.04). CONCLUSION: The prevalence of HIV, HBV and HCV seropositivity in jails suggests that information and education programs for inmates could be useful to reduce the spread of such infections

Extracellular Vesicles under Oxidative Stress Conditions: Biological Properties and Physiological Roles

Under physio-pathological conditions, cells release membrane-surrounded structures named Extracellular Vesicles (EVs), which convey their molecular cargo to neighboring or distant cells influencing their metabolism. Besides their involvement in the intercellular communication, EVs might represent a tool used by cells to eliminate unnecessary/toxic material. Here, we revised the literature exploring the link between EVs and redox biology. The first proof of this link derives from evidence demonstrating that EVs from healthy cells protect target cells from oxidative insults through the transfer of antioxidants. Oxidative stress conditions influence the release and the molecular cargo of EVs that, in turn, modulate the redox status of target cells. Oxidative stress-related EVs exert both beneficial or harmful effects, as they can carry antioxidants or ROS-generating enzymes and oxidized molecules. As mediators of cell-to-cell communication, EVs are also implicated in the pathophysiology of oxidative stress-related diseases. The review found evidence that numerous studies speculated on the role of EVs in redox signaling and oxidative stress-related pathologies, but few of them unraveled molecular mechanisms behind this complex link. Thus, the purpose of this review is to report and discuss this evidence, highlighting that the analysis of the molecular content of oxidative stress-released EVs (reminiscent of the redox status of originating cells), is a starting point for the use of EVs as diagnostic and therapeutic tools in oxidative stress-related diseases

Antioxidant systems and lymphocyte proliferation in the horse, sheep and dog

To better define the species-specific antioxidant systems and to ascertain the influence of the intracellular redox status on the immune system of different animal species, we determined lymphocyte glutathione peroxidase (GSHPx) activity, plasmatic glutathione levels (GSH) and the effect of H$_2$O$_2$ on the responsiveness of lymphocytes to proliferative stimuli. Among the three species considered, sheep presented the lowest plasmatic GSH and the highest lymphocyte GSHPx activity. On the contrary, dogs showed an inverted pattern (high GSH - low GSHPx). Horses displayed intermediate values for both parameters analysed. The effect of H$_2$O$_2$ on the proliferative capacity of lymphocytes was the same for all three species; the 200 $\mu$M dose in particular was strongly inhibiting. Each species, however, showed different rates of inhibition: sheep exhibited the highest sensitivity to the antiproliferative effect of H$_2$O$_2$. Our results confirmed that high H$_2$O$_2$ concentrations (200 $\mu$M) are noxious for the cellular functions of all animals; however this effect is mediated by a rigorously species-specific relationship between the intracellular reactive oxygen species (ROS) and the molecular systems involved in cell proliferation.Systèmes anti-oxydants et prolifération lymphocytaire chez le cheval, le mouton et le chien. Afin de définir l'influence de l'activité oxydo-réductrice intracellulaire sur le système immunitaire de différentes espèces animales, nous avons évalué l'activité de la glutathione peroxydase (GSHPx) lymphocytaire, de la glutathione (GSH) plasmatique et l'effet du peroxyde d'hydrogène (H$_2$O$_2$) sur la réponse des lymphocytes aux stimulations prolifératives. Parmi les trois espèces considérées, les moutons présentaient les plus faibles concentrations mineures en GSH plasmatique et l'activité du GSHPx lymphocytaire la plus élevée. Par contre, les chiens montraient un profil inverse (faible activité du GSHPx et concentration de GSH élevée). Les chevaux présentaient des valeurs moyennes pour les deux paramètres évalués. Les effets du H$_2$O$_2$ sur la capacité proliférative des lymphocytes ont été les mêmes dans les trois espèces étudiées, en particulier une dose de 200 $\mu$M s'est révélée fortement inhibitrice. Cependant, chaque espèce a montré des taux différents d'inhibition ; les moutons ont présenté une sensibilité maximale aux effets antiprolifératifs du H$_2$O$_2$. Nos résultats confirment que les concentrations élevées de H2O2 sont nocives pour la fonction cellulaire de tous les animaux. Par ailleurs, cet effet dépend, dans le contexte de la spécificité d'espèce, de la relation existant entre la concentration des dérivés réactifs à l'oxygène (ROS) intracellulaire et les systèmes moléculaires mis en jeu lors de la prolifération cellulaire