Effects of Nebivolol on Endothelial Gene Expression during Oxidative Stress in Human Umbilical Vein Endothelial Cells

The endothelium plays a key role in the development of atherogenesis and its inflammatory and proliferative status influences the progression of atherosclerosis. The aim of this study is to compare the effects of two beta blockers such as nebivolol and atenolol on gene expression in human umbilical vein endothelial cells (HUVECs) following an oxidant stimulus. HUVECs were incubated with nebivolol or atenolol (10 micromol/L) for 24 hours and oxidative stress was induced by the addition of oxidized (ox)-LDL. Ox-LDL upregulated adhesion molecules (ICAM-1, ICAM-2, ICAM-3, E-selectin, and P-selectin); proteins linked to inflammation (IL-6 and TNFalpha), thrombotic state (tissue factor, PAI-1 and uPA), hypertension such as endothelin-1 (ET-1), and vascular remodeling such as metalloproteinases (MMP-2, MMP-9) and protease inhibitor (TIMP-1). The exposure of HUVECs to nebivolol, but not to atenolol, reduced these genes upregulated by oxidative stress both in terms of protein and RNA expression. The known antioxidant properties of the third generation beta blocker nebivolol seem to account to the observed differences seen when compared to atenolol and support the specific potential protective role of this beta blocker on the expression of a number of genes involved in the initiation and progression of atherosclerosis

PIAZZE REALI E PIAZZE VIRTUALI VISUALIZZAZIONE INTERATTIVA DELLA RICOSTRUZIONE VIRTUALE DI PIAZZE FAMOSE COME STRUMENTO DI CONOSCENZA

<p>La piazza è un luogo di incontro, di confronto e di discussione, nel quale si creano nuovi rapporti o si rafforzano i rapporti esistenti tra le persone e il luogo stesso. Muoversi dentro una piazza consente di conoscere ‘fi sicamente’ la realtà nella quale stiamo agendo. La rappresentazione digitale di una piazza obbliga, invece, a dematerializzare la realtà e ad operare in una sua traduzione virtuale che consente di vivere l’esperienza di trovarsi in un luogo pur non essendoci. Tale esperienza non può sostituirsi al rapporto individuale che ciascuno di noi instaura con i luoghi reali, ma permette di ricreare con essi relazioni diverse, in accordo con i nostri sensi o addirittura ‘potenziandoli’, attraverso sensazioni nuove, ma comunque coinvolgenti. Un siffatto approccio può offrire un elevato contributo divulgativo perché può essere arricchito da numerose informazioni, su più livelli di conoscenza, fruibili mentre 'camminiamo' dentro la piazza ricostruita, proponendo dunque una capacità conoscitiva più intuitiva e diretta. Questo articolo si propone di esporre e discutere l’utilità e la fl essibilità di uso delle tecnologie virtuali attraverso la presentazione di una serie di ricostruzioni tridimensionali interattive di famose piazze, effettuate dal Laboratorio di Robotica Percettiva (PERCRO) della Scuola Superiore Sant’Anna di Pisa. Tali ricostruzioni, pur condividendo il medesimo linguaggio digitale, sono state utilizzate per un ampio ventaglio di scopi eterogenei, quali la pianifi cazione urbanistica, la comunicazione culturale, la promozione, l’informazione turistica, la fruizione artistica, nonché la divulgazione storica.</p><p> </p><p><strong>Virtual Square</strong></p><p>Squares  are  places  where  people  meet,  discuss,  confront  each other, where new relationships within the community are created or strengthened. Moving inside a square allows to physically know the reality in which we are acting. The digital representation of a square, instead, forces a dematerialization of the reality and makes us act in a virtual translation, allowing to experience a place with-out actually being there. This experience cannot replace the indi-vidual relationship established with real places, though it allows to create new and different connections with them, in agreement with our senses or even augmenting them through new involving sensa-tions. Such an approach may offer a relevant contribution for communication and dissemination, as it can provide several information on different levels of knowledge, accessible, while we ‘walk’ inside the reconstructed square, in a more direct and intuitive way. This article aims to describe and discuss the usefulness and the fl exibility of virtual technologies through the presentation of a series of 3D reconstructions of famous squares, produced by the Perceptual Ro-botics Laboratory (PERCRO) of Scuola Superiore Sant’Anna of Pisa. These reconstructions, although sharing the same digital language, have been used for a wide range of heterogeneous purposes, such as urban planning, cultural communication, promotion, tourist information and education.</p&gt

automatic creation of a virtual augmented gallery based on user defined queries on online public repositories

Virtual museums are becoming increasingly popular, especially thanks to the recent spread of low-cost immersive technologies enabling a richer technology-based cultural offer. However, creating a virtual museum commonly requires a lot of effort, especially if a certain visual quality is required

Repeated administration of the spasmolytic otilonium bromide counteracts functional and neurotransmitters’ changes in the colon of rats underwent to wrap restraint stress

Otilonium bromide (OB) is a quaternary ammonium derivative successfully used for the treatment of irritable bowel syndrome (IBS). Several in vitro experiments in human and rat colon demonstrated its spasmolytic capability due to the block of muscarinic and tachykinin receptors and L-type Ca2+ channels. Moreover, in vivo OB administrations showed interesting interaction with the enteric nervous system in healthy rats (1). The wrap restrain stress (WRS) is considered an adequate model of psychosocial stressor, able to induce most of the IBS signs and symptoms. WRS leads to important changes in the enteric neurotransmitters of rat colon, as recently demonstrated (2). Consequently, we chose this animal model to investigate whether a repeated, oral treatment with OB prevented the functional and neurotransmitters’ changes reported in rats underwent to WRS. The results obtained by using multiple experimental approaches (in vivo colonic functional evaluations, routine histology, immunohistochemistry and western blot) showed that OB is able to counteract most of the morphological changes caused by WRS in the colonic wall. In particular, the drug prevents the decrease in SP-, NK1r-, nNOS-, VIP- and S100β-immunoreactivity (IR) and the increase in CGRP- and CRF1r-IR detected in WRS rats. On the contrary, OB does not interfere with the mild mucosal inflammation and does not affect the increase in CRF2r-immunoreactive neurons observed in WRS rats. Moreover, OB per se increases the muscarinic receptor 2 expression in the muscle wall and decreases the number of the myenteric ChAT-immunoreactive neurons. Functional findings show a significantly reduction in the number of spontaneous abdominal contraction in OB treated rats. The ability of OB to block L-type Ca2+ channels, also expressed by enteric neurons, might explain the drug efficacy in preventing excessive neuronal response to stress. This work was supported by grants from Menarini I.F.R

The wrap partial restrain stress, an animal model of the irritable bowel syndrome: immunohistochemical and functional characterization

Several animal models have been proposed to mimic the human irritable bowel syndrome (IBS) all based on two etio-pathogenic hypotheses: infection and stress, both responsible for the development of a local inflammation. We investigated the wrap partial restrain stress (WRS) animal model with the aim to evaluate its validity in understanding the human IBS. Male Wistar rats were used and WRS was maintained for 2h. Abdominal contractions (AC) were recorded by a distension balloon in the colon-rectum. The number of faecal pellets and their total weight were determined. Colonic specimens from both groups were examined by routine histology, immunohistochemistry and western blot (WB). WRS animals were characterized by: 1) a statistically significant increase in the number of AC and in the mean number and mean weight of faecal pellets; 2) the presence of large clusters of mononucleated cells and a significant increase in eosinophilic granulocytes and mast cells in the mucosa; 3) reduction of GLP1r-immunoreactivity (IR) located at the basolat- eral periphery and the Golgi level of the cells of the glandular funds; 4) an increase in CGRP-IR in the lamina propria; 5) no significant difference in the muscle wall for Cav1, L- type Ca+2-channels, Mr2, NK1r and NK2r; 6) a significant decrease in the myenteric and a significant increase in the submucous NK1-IR neuron number; 7) a significant decrease in Substance P-IR in the myenteric plexus and muscle coat; 8) a significant decrease in myenteric and submucous nNOS-IR neuron number; 9) no difference in ChAT-IR neurons of both enteric plexuses; 10) a reduction in S-100-IR in the entire colonic wall; 11) no difference in the total number of neurons evaluated by the pan-neuronal marker PGP 9.5; 12) no change of all the ICC populations. The functional data are in favor of a lowering in the colonic wall distention threshold; the morphological results obtained in the lamina propria demonstrate the presence of a local inflammation, particularly intense at the level of the mucosa. Both of these findings agree with the hypothesis that inflammation might have a main role in the insurgence and maintenance of the typical IBS symptoms and support the validity of our WRS model. Moreover, while the smooth muscle cells do not show any significant variation, numerous and consistent changes in the excitatory, inhibitory and NK1r-IR neurons are detected

Seed storage allergens tackled via next-generation research assistant

The expanding consumption of plant proteins in the diet to overcome the environmental issues associated with animal proteins is increasing the incidence of food-induced allergic reactions. One of the 21st-century research drivers in agriculture sciences is the development and validation of concrete approaches for modulating the expression of allergenic proteins in crops before harvesting. The increasing incidence of plant food allergies is primarily induced by seed storage proteins that clinicians are experiencing recently because of the more predominant use of plant-derived proteins in the food industry. Increased availability of high-throughput technologies has generated an ever-growing number of omics data, allowing us to have better structural knowledge of SSPs and molecular properties that can inform the allergenicity assessment. The recent systems for targeted genome engineering, without double-strand DNA breaks, allow the introduction of precise modifications directly into commercial plant species. Artificial intelligence is significantly transforming scientific research across every stage, assisting scientists, processing large-scale data, making predictions, automating tasks. During this epochal change, marked by the encounter between artificial intelligence and synthetic biology, a next-generation research assistant (NGA) is coming alive. Here, we propose a new conceptual vision to facilitate and speed up the editing of cross-reactivity sites to obtain hypoallergenic cultivars and avoid pleiotropic effects. Finally, we discuss the potential applications of this new way to conceive the research. NGA may be undoubtedly capable of managing the evolution of SPP allergies through the prediction of novel epitopes, as well as the prediction of immunological response mechanisms

Ascorbic acid reduces Ropivacaine-induced myotoxicity in cultured human osteoporotic skeletal muscle cells

Background: Osteoporosis is a worldwide health issue. Loss of bone mass is a potential risk factor for fragility fractures, and osteoporotic fractures place a considerable burden on society. Bone and muscle represent a functional unit in which the two tissues are intimately interconnected. Ropivacaine is a potent local anesthetic used in clinical practice for intraoperative anesthesia and postoperative pain management, in particular for hip surgery. When injected, Ropivacaine can diffuse locally through, in particular in surrounding skeletal muscle tissue, causing dose-dependent cytotoxicity, oxidative stress and myogenesis impairment. Based on those evidences, we focused our attention on Ropivacaine-induced cytotoxicity on cultured human myoblasts. Methods: Primary human myoblasts and myotubes from healthy subjects, osteoarthritic and osteoporotic patients (OP) were cultured in the presence of Ropivacaine. In some experiments, ascorbic acid (AsA) was added as a potent antioxidant agent. Cell viability and ROS levels were evaluated to investigate the myotoxic activity and Real-Time PCR and Western blot analysis carried out to investigate the expression of proliferation and myogenic markers. Results: A dose-dependent decrease of cell viability was observed after Ropivacaine exposure in both OP myoblasts and myotubes cultures, whereas those effects were not observed in the presence of Propofol, a general anesthetic. The adding of AsA reduced Ropivacaine negative effects in OP myoblast cultures. In addition, Ropivacaine exposure also increased ROS levels and upregulated Nox4 expression, an enzyme primarily implicated in skeletal muscle ROS generation. AsA treatment counteracted the oxidant activity of Ropivacaine and partially restored the basal condition in cultures. Positive myogenic markers, such as MyoD and Myf5, were downregulated by Ropivacaine exposure, whereas myostatin, a negative regulator of muscle growth and differentiation, was upregulated. The phenotypic deregulation of myogenic controllers in the presence of Ropivacaine was counteracted by AsA treatment. Conclusions: Our findings highlight the oxidative stress-mediated myotoxic effect of Ropivacaine on human skeletal muscle tissue cell cultures, and suggest treatment with AsA as valid strategy to mitigate its negative effects and allowing an ameliorated functional skeletal muscle recovery in patients undergoing hip replacement surgery for osteoporotic bone fracture

Chronic treatment with otilonium bromide affects the tachykinergic and nitrergic systems in the rat colon

Otilonium bromide (OB), a quaternary ammonium derivative used for the treatment of intestinal motility disorders such as the irritable bowel syndrome (IBS). It exerts several actions, among which the ability to bind to the neurokin-2 receptor (NK2r) inhibiting NK2r mediated contraction and, in the human colon, NK2r internalization in the smooth muscle cells (SMC) (Cipriani et al., 2011). Substance P (SP) is an excitatory neurotransmitter that, interacting mainly with the neurokinin-1 receptor (NK1r), can stimulate bowel motility by SMC direct activation or inhibit it by an indirect action through enteric neural circuits. In an IBS rat model, the increase in NK1rmediated colonic motor response was associated to a decrease in the nitrergic activity. On these basis, we tested whether OB modifies NK1r, NK2r, SP and neuronal nitric oxide synthase (nNOS) expression in rat colon after chronical administration of the drug (2 or 20mg/Kg/daily) for 10 or 30 days. At the end of the treatments, specimens of proximal colon were collected and the expression of NK1r, NK2r, SP and neurogenic and myogenic nNOS were evaluated by immunohistochemistry and Western blot. Our data show that SP expression was significantly decreased in 10 and 30 days treated rats in myenteric ganglia and, in 30 days treated rats, also in the intramuscular nerve fibres. No quantitative change of the two NKr was observed, whereas, after 30 days, the NK1r was concentrated in the SMC cytoplasm. In parallel, the neurogenic nNOS expression increased and reached the significance after 30 days of treatment; the myogenic nNOS expression increased, but these increase reached the significance only at 10 days. Our findings suggest that the main target of the OB chronically administered is the NO-mediated system that is stimulated earlier at the muscular level, later at the neuronal level. We interpret the systemic decrease in the SP expression as consequence of the potentiated NO availability in the ganglia and muscle coat. If true, the late concentration of NK1r in the cytoplasm could represent an attempt of the SMC to overcome the deficit of its main ligand SP

Inner and outer portions of colonic circular muscle: ultrastructural and immunohistochemical changes in rat chronically treated with Otilonium Bromide

Rat colonic circular muscle layer (cml), main target of otilonium bromide (OB) spasmolytic activity [1], is subdivided in an inner (icl) and an outer (icl) portion. The icl is particularly rich in organelles such as caveolae, smooth endoplasmic reticulum, mitochondria [2]. The expression of specific markers in comparison with the ocl was investigated. The possible changes of these organelles and related markers, and of muscarinic receptors (Mr2) were studied after OB chronic exposition (2-20mg/kg for 10 or 30 days). Colon specimens were processed for electron microscopy, immunohistochemistry and western blot. In colonic strips the contractility response to Mr2 agonist was investigated. The results show that in controls the icl displays a higher expression of Caveolin-1 and Mr2, but not of eNOS, calreticulin and calsequestrin, compared to ocl. Chronic OB treatment caused similar ultrastructural and immunohistochemical changes in both portions. Organelles, Caveolin-1 and eNOS were increased at 10 days; Mr2 expression and muscle contractility induced by metacholine were increased at 30 days. Our findings 1) provide new information on the immunohistochemical properties of the icl suggesting a distinct role for this portion in colonic motility; 2) demonstrate that chronically administered OB interferes with cell structures and molecules responsible for calcium handling and storage, and modifies cholinergic transmission