This pilot study aimed at investigating how salivary oxytocin levels are affected by human interaction and isolation in eight guide dogs (six Labrador retrievers and two golden retrievers; four males and four females, 21.87 ± 1.36 months old) just before assignment to the blind person. Each dog engaged, at one-week intervals, in a positive (5 min of affiliative interaction with their trainer) and a negative (5 min of isolation) condition. Saliva samples used for Enzyme Immunoassay (EIA) quantification of salivary oxytocin were collected before and immediately after both experimental conditions. In order to assess potential hypothalamic pituitary adrenal (HPA) axis activation that could have affected oxytocin levels, saliva samples were collected 15 min after both experimental conditions for EIA quantification of salivary cortisol and a behavioral assessment was performed during the negative condition. The results were compared using the Wilcoxon test (p < 0.05). Oxytocin concentrations showed a statistically significant increase after the positive interaction (p = 0.036) and no difference after the negative one (p = 0.779). Moreover, no difference (p = 0.263) was found between the cortisol concentrations after each experimental condition and no signs of distress were observed during the isolation phase. These preliminary findings support the hypothesis that stroking dogs has positive effects on their emotional state independently of hypothalamic pituitary adrenal axis activation.

SummaryBrown hare (Lepus europaeus) populations in Europe have declined through decades due to several, but not clear yet, factors. Parasite infections and diseases are some of the causes that directly affected the survival and breeding rates of animal population.A study on the endoparasites of 70 hares (37 hunted free-living hares, and 33 bred on farms hares) was performed between 2015 - 2017 in the province of Grosseto (central Italy), an area where the impact of parasites in the hare population has never been investigated. During necroscopic analysis of hunted hares the following helminthes were found: Trichostrongylus retortaeformis (87.1 %), Passalurus ambiguus (12.9 %) and Andrya spp. (6.4 %) in the intestinal tract, Protostrongylus cuniculorum (8.3 %) in lungs and Dicrocoelium dendriticum (16.7 %) in livers. The prevalences of the intestinal helminthes in bred hares were: 12.1 % for Passalurus ambiguus and 3 % for Trichostrongylus retortaeformis. The coprological analysis showed prevalences of 64.9 % for coccidia in the 37 hunted hares and 45.5 % in the 33 bred hares. The relationship between the intensities of parasitic infections and body weight was evaluated.The results of the present study in the Grosseto area indicate that free-living hares have few species of parasites and that the intensities of parasitic infection did not affect their general condition and health, suggesting that endoparasites played no detectable role in the dynamics of this hare population.

In mammals, serotonin (5-HT) levels depend on the availability of tryptophan (TRP). Low 5-HT concentrations have been linked to behavioural disorders in dogs. This study aimed at investigating possible differences in dogs’ serum TRP and 5-HT concentrations according to their behavioural response to a potentially stressful procedure. Thirty-nine physically healthy shelter dogs, 15 females and 24 males, mean age = 5.6 years, were categorized by a certified veterinary behaviourist according to their behavioural response to medical examination and blood collection, in: relaxation, stress signals, tension without growling, tension with growling, escape attempts, and aggression attempts. Extraction and quantification of 5-HT and TRP were performed using a HLPC method. Data were statistically analysed, applying Chi-square and Spearman tests. Results showed no significant difference in TRP (χ2 = 2.084, p = 0.555) nor 5-HT (χ2 = 0.972, p = 0.808) serum concentrations among different categories of dogs; however, some categories were underrepresented (relaxation = 20.5%, stress signals = 30.8%, tension without growling = 43.6%, tension with growling = 5.1%, escape attempts = 0%, aggression attempts = 0%). No correlation between serum TRP and 5-HT concentrations was found (ρ = 0.086, p = 0.602). Serum 5-HT levels do not seem to be associated with dogs’ behavioural response to a stressful situation nor with serum TRP concentrations. The relationship between serum TRP and 5-HT concentrations and behaviour needs further research.

The receptor for advanced glycation end products (RAGE) is implicated in diabetes and obesity complications, as well as in breast cancer (BC). Herein, we evaluated whether RAGE contributes to the oncogenic actions of Insulin, which plays a key role in BC progression particularly in obese and diabetic patients. Analysis of the publicly available METABRIC study, which collects gene expression and clinical data from a large cohort (n = 1904) of BC patients, revealed that RAGE and the Insulin Receptor (IR) are co-expressed and associated with negative prognostic parameters. In MCF-7, ZR75 and 4T1 BC cells, as well as in patient-derived Cancer-Associated Fibroblasts, the pharmacological inhibition of RAGE as well as its genetic depletion interfered with Insulin-induced activation of the oncogenic pathway IR/IRS1/AKT/CD1. Mechanistically, IR and RAGE directly interacted upon Insulin stimulation, as shown by in situ proximity ligation assays and coimmunoprecipitation studies. Of note, RAGE inhibition halted the activation of both IR and insulin like growth factor 1 receptor (IGF-1R), as demonstrated in MCF-7 cells KO for the IR and the IGF-1R gene via CRISPR-cas9 technology. An unbiased label-free proteomic analysis uncovered proteins and predicted pathways affected by RAGE inhibition in Insulin-stimulated BC cells. Biologically, RAGE inhibition reduced cell proliferation, migration, and patient-derived mammosphere formation triggered by Insulin. In vivo, the pharmacological inhibition of RAGE halted Insulin-induced tumor growth, without affecting blood glucose homeostasis. Together, our findings suggest that targeting RAGE may represent an appealing opportunity to blunt Insulin-induced oncogenic signaling in BC.