Aliskiren inhibits prorenin-induced human aortic smooth muscle cell migration

Background: In the present study, we investigated the potential effect of aliskiren on smooth muscle cell (SMC) migration in response to prorenin. Methods: Cultured human SMCs were incubated with angiotensinogen (ANG) (1.5 10-7M) and increasing concentrations of aliskiren (10-610-5M). After 24 h, SMC migration was assessed by Boydens chamber chemotactic assay using prorenin as chemotactic factor (10-8M). The effect of aliskiren on RhoA and Rac activity was also determined by G-LISA assay and the lamellipodia formation by rhodaminephalloidin staining. Changes in cell morphology were recorded in real-time using the iCelligence system. Results: Aliskiren determined, at 10-5M, a significant inhibition of SMC migration induced by prorenin (-66.418.1%; p > 0.05), while no significant effect was observed when PDGF-BB was utilized as chemotactic agent. Aliskiren also reduced Rac-GTP levels in response to prorenin (-54.2\ub15.4%) without affecting the RhoA-GTP levels. Finally, aliskiren inhibited both the lamellipodia formation and morphological changes induced by prorenin with no significant effect on PDGF-BB activity. Conclusions: Taken together, we provide the first evidence of the inhibitory action of aliskiren on SMC migration induced by prorenin

Modulation of the intrinsic neuronal excitability by multifunctional liposomes tailored for the treatment of alzheimer’s disease

Purpose: Nanotechnologies turned out to be promising in the development of diagnostic and therapeutic approaches toward neurodegenerative disorders. However, only a very scant number of nanodevices until now proved to be effective on preclinical animal models. Although specific tests in vivo are available to assess the potential toxicity of these nanodevices on cognitive functions, those to evaluate their biosafety in vitro on neurons are still to be improved. Materials and methods: We utilized the patch-clamp technique on primary cultures of cortical neural cells isolated from neonatal rats, aiming to evaluate their electrical properties after the incubation with liposomes (mApoE-PA-LIPs), previously proved able to cross the blood\u2013brain barrier and to be effective on mouse models of Alzheimer\u2019s disease (AD), both in the absence and in the presence of \u3b2-amyloid peptide oligomers. Results: Data show a high degree of biocompatibility, evaluated by lactate dehydrogenase (LDH) release and MTT assay, and the lack of cellular internalization. After the incubation with mApoE-PA-LIPs, neuronal membranes show an increase in the input resistance (from 724.14\ub176 M\u3a9 in untreated population to 886.06\ub186 M\u3a9 in the treated one), a reduction in the rheobase current (from 29.6\ub13 to 24.2\ub13 pA in untreated and treated, respectively), and an increase of the firing frequency, consistent with an ultimate increase in intrinsic excitability. Data obtained after co-incubation of mApoE-PA-LIPs with \u3b2-amyloid peptide oligomers suggest a retention of liposome efficacy. Conclusion: These data suggest the ability of liposomes to modulate neuronal electrical properties and are compatible with the previously demonstrated amelioration of cognitive functions induced by treatment of AD mice with liposomes. We conclude that this electrophysiological approach could represent a useful tool for nanomedicine to evaluate the effect of nanoparticles on intrinsic neuronal excitability

New FTY720-docetaxel nanoparticle therapy overcomes FTY720-induced lymphopenia and inhibits metastatic breast tumour growth

Purpose: Combining molecular therapies with chemotherapy may offer an improved clinical outcome for chemoresistant tumours. Sphingosine-1-phosphate (S1P) receptor antagonist and sphingosine kinase 1 (SK1) inhibitor FTY720 (FTY) has promising anticancer properties, however, it causes systemic lymphopenia which impairs its use in cancer patients. In this study, we developed a nanoparticle (NP) combining docetaxel (DTX) and FTY for enhanced anticancer effect, targeted tumour delivery and reduced systemic toxicity. Methods: Docetaxel, FTY and glucosamine were covalently conjugated to poly(lactic-co-glycolic acid) (PLGA). NPs were characterised by dynamic light scattering and electron microscopy. The cellular uptake, cytotoxicity and in vivo antitumor efficacy of CNPs were evaluated. Results: We show for the first time that in triple negative breast cancer cells FTY provides chemosensitisation to DTX, allowing a four-fold reduction in the effective dose. We have encapsulated both drugs in PLGA complex NPs (CNPs), with narrow size distribution of ~ 100 nm and excellent cancer cell uptake providing sequential, sustained release of FTY and DTX. In triple negative breast cancer cells and mouse breast cancer models, CNPs had similar efficacy to systemic free therapies, but allowed an effective drug dose reduction. Application of CNPs has significantly reversed chemotherapy side effects such as weight loss, liver toxicity and, most notably, lymphopenia. Conclusions: We show for the first time the DTX chemosensitising effects of FTY in triple negative breast cancer. We further demonstrate that encapsulation of free drugs in CNPs can improve targeting, provide low off-target toxicity and most importantly reduce FTY-induced lymphopenia, offering potential therapeutic use of FTY in clinical cancer treatment

Effect of aliskiren on (pro)renin receptor expression and activity: in vitro determination of TGF-β, PAI-1 and type I collagen expression and smooth muscle cell migration

The recent discovery of a specific receptor for renin/prorenin (PRR) has added new interest to the pharmacological actions of aliskiren, the first direct renin inhibitor. In the present study we investigated the effect of aliskiren on PRR expression and activity in cultured human smooth muscle cells (SMCs). Co-incubation of SMCs with angiotensinogen (ANG) (1.5 7 10-7 M) and prorenin (10-8\u201310-7 M) resulted in an efficient production of angiotensin I, almost completely inhibited by 10-5 M aliskiren (-86.0 \ub1 14.0%). A 24 h incubation with aliskiren (10-6\u201310-5 M) resulted in a concentration-dependent reduction of PRR mRNA levels (IC50 4.6 7 10-6 M) and the cell surface expression of PRR (IC50 9.1 7 10-6 M). The lower levels of PRR were associated with a reduced expression of TGF-\u3b2, PAI-1 and type I collagen mRNA. The effect of prorenin on SMC migration was also investigated. Prorenin induced SMC migration in a dose-dependent manner, as assessed by Boyden chamber chemotaxis assay. The knockdown of PRR by small interfering RNA completely inhibited the migratory response to prorenin, demonstrating that the chemotactic action of prorenin is mediated by the PRR. Prorenin increased the intracellular levels of both RhoA-GTP (+47.8%) and Rac1-GTP (+36.7%) through PRR. A 24 h incubation with aliskiren (10-6 - 2.5x10-5 M) did not affect SMC migration and RhoA-GTO levels in response to prorenin. In conclusion aliskiren elicits a direct pharmacological action on PRR expression and its signaling pathway in SMCs, affecting gene expression of TGF-\u3b2, PAI-1 and type I collagen. However, this action is not sufficient to significantly reduced the chemotactic action of prorenin on SMCs. These results, although obtained from in vitro analysis, may help to better define the pharmacological properties of aliskiren on atherosclerosis. The present work was supported by a grant from Novartis Farma S.p.A., Origgio, Italy

Enhanced recovery pathways in thoracic surgery from Italian VATS group: Nursing care program

Enhanced recovery after surgery (ERAS) is an interprofessional program that can lead to hastened patient recovery and reduced time in hospital. Nursing staff play a key role in the implementation of enhanced recovery protocols. This issue focalizes the role of nurses in ERAS program for patients submitted to Thoracic Surgery, in particular for cases of major lung resection performed by a minimally invasive surgical approach (VATS, video assisted thoracic surgery)