Possibili relazioni tra disturbi comportamentali su base ansiosa nel cane e neurotrasmettitori

La risposta di stress diviene problematica quando un animale \ue8 incapace di controllare o di sottrarsi a una situazione tramite un\u2019appropriata risposta. La risposta ansiosa non solo inizia automaticamente ogniqualvolta il soggetto incontra uno stimolo particolare, non prevedibile o che presagisce una situazione non prevedibile, ma provoca spesso un problema di generalizzazione nei confronti di stimoli simili: in questi casi la risposta di stress diviene cronica e gli animali manifestano risposte comportamentali inappropriate o eccessive al fine di ridurre l\u2019effetto deleterio di una reazione prolungata. Relazioni tra i livelli di alcuni neurotrasmettitori e problemi comportamentali su base ansiosa sono state riscontrate in numerose specie. In particolare si riportano i principali risultati di una ricerca, condotta su 20 cani \u201cproblematici\u201d e 13 soggetti controllo, in cui sono state analizzate le relazioni tra i livelli di alcuni neurotrasmettitori e diverse problematiche comportamentali ansiose e aggressive su base ansiosa. I livelli plasmatici di dopamina (DA) e serotonina (5HT) risultano significativamente pi\uf9 elevati nei soggetti ansiosi rispetto al gruppo controllo. I livelli plasmatici degli altri neurotrasmettitori sono invece simili nei due gruppi, cos\uec come i livelli piastrinici di noradrenalina (NE) e dopamina (DA). \uc8 stato osservato un trend di concentrazione serotoninica nelle piastrine pi\uf9 elevata nei soggetti di controllo rispetto al gruppo dei \u201cproblematici\u201d

Automated Large-Scale Production of Paclitaxel Loaded Mesenchymal Stromal Cells for Cell Therapy Applications

Mesenchymal stromal cells (MSCs) prepared as advanced therapies medicinal products (ATMPs) have been widely used for the treatment of different diseases. The latest developments concern the possibility to use MSCs as carrier of molecules, including chemotherapeutic drugs. Taking advantage of their intrinsic homing feature, MSCs may improve drugs localization in the disease area. However, for cell therapy applications, a significant number of MSCs loaded with the drug is required. We here investigate the possibility to produce a large amount of Good Manufacturing Practice (GMP)-compliant MSCs loaded with the chemotherapeutic drug Paclitaxel (MSCs-PTX), using a closed bioreactor system. Cells were obtained starting from 13 adipose tissue lipoaspirates. All samples were characterized in terms of number/viability, morphology, growth kinetics, and immunophenotype. The ability of MSCs to internalize PTX as well as the antiproliferative activity of the MSCs-PTX in vitro was also assessed. The results demonstrate that our approach allows a large scale expansion of cells within a week; the MSCs-PTX, despite a different morphology from MSCs, displayed the typical features of MSCs in terms of viability, adhesion capacity, and phenotype. In addition, MSCs showed the ability to internalize PTX and finally to kill cancer cells, inhibiting the proliferation of tumor lines in vitro. In summary our results demonstrate for the first time that it is possible to obtain, in a short time, large amounts of MSCs loaded with PTX to be used in clinical trials for the treatment of patients with oncological diseases

Gemcitabine-releasing mesenchymal stromal cells inhibit in vitro proliferation of human pancreatic carcinoma cells

BACKGROUND AIMS: Pancreatic cancer (pCa) is a tumor characterized by a fibrotic state and associated with a poor prognosis. The observation that mesenchymal stromal cells (MSCs) migrate toward inflammatory micro-environments and engraft into tumor stroma after systemic administration suggested new therapeutic approaches with the use of engineered MSCs to deliver and produce anti-cancer molecules directly within the tumor. Previously, we demonstrated that without any genetic modifications, MSCs are able to deliver anti-cancer drugs. MSCs loaded with paclitaxel by exposure to high concentrations release the drug both in vitro and in vivo, inhibiting tumor proliferation. On the basis of these observations, we evaluated the ability of MSCs (from bone marrow and pancreas) to uptake and release gemcitabine (GCB), a drug widely used in pCa treatment. METHODS: MSCs were primed by 24-h exposure to 2000 ng/mL of GCB. The anti-tumor potential of primed MSCs was then investigated by in vitro anti-proliferation assays with the use of CFPAC-1, a pancreatic tumor cell line sensitive to GCB. The uptake/release ability was confirmed by means of high-performance liquid chromatography analysis. A cell-cycle study and secretome evaluation were also conducted to better understand the characteristics of primed MSCs. RESULTS: GCB-releasing MSCs inhibit the growth of a human pCa cell line in vitro. CONCLUSIONS: The use of MSCs as a "trojan horse" can open the way to a new pCa therapeutic approach; GCB-loaded MSCs that integrate into the tumor mass could deliver much higher concentrations of the drug in situ than can be achieved by intravenous injection

Human Olfactory Bulb Neural Stem Cells (Hu-OBNSCs) Can Be Loaded with Paclitaxel and Used to Inhibit Glioblastoma Cell Growth

Exploitation of the potential ability of human olfactory bulb (hOB) cells to carry, release, and deliver an effective, targeted anticancer therapy within the central nervous system (CNS) milieu remains elusive. Previous studies have demonstrated the marked ability of several types of stem cells (such as mesenchymal stem cells (MSCs) to carry and release different anti-cancer agents such as paclitaxel (PTX). Herein we investigate the ability of human olfactory bulb neural stem cells (Hu-OBNSCs) to carry and release paclitaxel, producing effective cytotoxic effects against cancer cells. We isolated Hu-OBNSCs from the hOB, uploaded them with PTX, and studied their potential cytotoxic effects against cancer cells in vitro. Interestingly, the Hu-OBNSCs displayed a five-fold increase in their resistance to the cytotoxicity of PTX, and the PTX-uploaded Hu-OBNSCs were able to inhibit proliferation and invasion, and to trigger marked cytotoxic effects on glioblastoma multiforme (GBM) cancer cells, and Human Caucasian fetal pancreatic adenocarcinoma 1 (CFPAC-1) in vitro. Despite their ability to resist the cytotoxic activity of PTX, the mechanism by which Hu-OBNSCs acquire resistance to PTX is not yet explained. Collectively our data indicate the ability of the Hu-OBNSCs to resist PTX, and to trigger effective cytotoxic effects against GBM cancer cells and CFPAC-1. This indicates their potential to be used as a carrier/vehicle for targeted anti-cancer therapy within the CNS

Transmission of Chronic Wasting Disease Identifies a Prion Strain Causing Cachexia and Heart Infection in Hamsters

Chronic wasting disease (CWD) is an emerging prion disease of free-ranging and captive cervids in North America. In this study we established a rodent model for CWD in Syrian golden hamsters that resemble key features of the disease in cervids including cachexia and infection of cardiac muscle. Following one to three serial passages of CWD from white-tailed deer into transgenic mice expressing the hamster prion protein gene, CWD was subsequently passaged into Syrian golden hamsters. In one passage line there were preclinical changes in locomotor activity and a loss of body mass prior to onset of subtle neurological symptoms around 340 days. The clinical symptoms included a prominent wasting disease, similar to cachexia, with a prolonged duration. Other features of CWD in hamsters that were similar to cervid CWD included the brain distribution of the disease-specific isoform of the prion protein, PrPSc, prion infection of the central and peripheral neuroendocrine system, and PrPSc deposition in cardiac muscle. There was also prominent PrPSc deposition in the nasal mucosa on the edge of the olfactory sensory epithelium with the lumen of the nasal airway that could have implications for CWD shedding into nasal secretions and disease transmission. Since the mechanism of wasting disease in prion diseases is unknown this hamster CWD model could provide a means to investigate the physiological basis of cachexia, which we propose is due to a prion-induced endocrinopathy. This prion disease phenotype has not been described in hamsters and we designate it as the ‘wasting’ or WST strain of hamster CWD

Biological and hydraulic monitoring of sediment flushing from an alpine reservoir

The first results of the biological and hydraulic monitoring programme carried out during the sediment flushing of an Alpine Italian reservoir are presented. The flushing and the monitoring activities were planned and executed by several private and public subjects, including the hydroelectric company in charge of the reservoir management (AEM S.p.A.), local environmental agencies, an angler association, a company of environmental consultancy (Graia S.r.l.), the Insubria university. The Italian normative requires that the sediment removal from a reservoir should follow a Reservoir Management Plan compiled by the reservoir manager (which is also in charge for the execution of the flushing operation). Difficulties arise in planning such operations since no threshold limits for the suspended solid (SS) concentration of the outflow waters are defined at National level. For this reason the present research was addressed to provide experimental evidence of the SS effects on the aquatic biota present in the creek downstream of the flushed reservoir, aiming at establish ecologically-based threshold limits of SS concentration. A second objective of the research was to verify how much the effluent SS concentration could be controlled by means of technical operations for flushing management

Determination of 5-chloro-7-iodo-8-quinolinol (clioquinol) in plasma and tissues of hamsters by high-performance liquid chromatography and electrochemical detection

This paper describes a method of determining clioquinol levels in hamster plasma and tissue by means of HPLC and electrochemical detection. Clioquinol was separated on a Nucleosil C18 300 x 3.9 mm I.D. 7 \ub5m column at 1 ml/min using a phosphate/citrate buffer 0.1 M (400 ml) with 600 ml of a methanol:acetonitrile (1:1, v/v) mobile phase. The retention times of clioquinol and the IS were respectively 11.6 and 8.1 min; the quantitation limit (CV>8%) was 5 ng/ml in plasma and 10 ng/ml in tissues. The intra- and inter-assay accuracies of the method were more than 95%, with coefficients of variation between 3.0% to 7.7%, and plasma and tissue recovery rates of 72-77%. There was a linear response to clioquinol 5-2000 ng/ml in plasma, and 10-1000 ng/g in tissues. The method is highly sensitive and selective, makes it possible to study the pharmacokinetics of plasma clioquinol after oral administration and the distribution of clioquinol in tissues, and could be used to monitor plasma clioquinol levels in humans

Levodopa and 3-OMD levels in Parkinson patients treated with Duodopa

We studied 19 patients (14 men, 5 women, Hoen and Yahr>3) with advanced Parkinson's disease(PD) attending the Parkinson Institute,Milan, whose motor fluctuation and dyskinesia were not contrlled by oral medication. After all oral PD medications had been withdrawn, they received a duodenal levodopa infusions (Duodopa, Solvay Pharmaceutical) for 14h/day through a transabdominal port; levodopa boluses were administered in the morning and during "off" peeriod. The patient were evaluated by means of the UPDRS in the morning ("off") and 60-120 min after the infusion ("on") at baseline and for a mean follow-up of 13.5+12.5 months(up to 36 months in 10 patients:). Levodopa (L-DOPA) and its metabolites were determined by HPLC with electrochemical detection. L-Dopa concentration tended to higher in the afternoon (2008+ 345 vs 1713+ 274 ng/ml) and correlated with the daily dose. O-methyldopa (OMD) levels correlated with L-Dopa levels, and the OMD/L-DOPA ratios were stable over the day. There was a relationship between the decreasing UPDRS III scores and decreasing OMD/L-DOPA ratios.Dyskinesia (UPDRS IV, items 32-34) showed a clear improvement over time but there was no clear relationship with L-DOPA and OMD levels, or the OMD/L-DOPA ratio. The L-DOPA/dose ratio was stable over time, whereas OMD levels and the OMD/L-DOPA ratio decresed. It is conceivable that continuos infusion decreases metabolism possibly due to a reduction in methyl donor availability, as demonstrated by the increase in total homocysteine levels. Our results do not support the development of tolerance even after several months of continuous infusion, and indicate that pharmacodynamic factors play a role in afternoon off periods