Endothelial cells, endoplasmic reticulum stress and oxysterols

Oxysterols are bioactive lipids that act as regulators of lipid metabolism, inflammation, cell viability and are involved in several diseases, including atherosclerosis. Mounting evidence linked the atherosclerosis to endothelium dysfunction; in fact, the endothelium regulates the vascular system with roles in processes such as hemostasis, cell cholesterol, hormone trafficking, signal transduction and inflammation. Several papers shed light the ability of oxysterols to induce apoptosis in different cell lines including endothelial cells. Apoptotic endothelial cell and endothelial denudation may constitute a critical step in the transition to plaque erosion and vessel thrombosis, so preventing the endothelial damaged has garnered considerable attention as a novel means of treating atherosclerosis. Endoplasmic reticulum (ER) is the site where the proteins are synthetized and folded and is necessary for most cellular activity; perturbations of ER homeostasis leads to a condition known as endoplasmic reticulum stress. This condition evokes the unfolded protein response (UPR) an adaptive pathway that aims to restore ER homeostasis. Mounting evidence suggests that chronic activation of UPR leads to cell dysfunction and death and recently has been implicated in pathogenesis of endothelial dysfunction. Autophagy is an essential catabolic mechanism that delivers misfolded proteins and damaged organelles to the lysosome for degradation, maintaining basal levels of autophagic activity it is critical for cell survival. Several evidence suggests that persistent ER stress often results in stimulation of autophagic activities, likely as a compensatory mechanism to relieve ER stress and consequently cell death. In this review, we summarize evidence for the effect of oxysterols on endothelial cells, especially focusing on oxysterols-mediated induction of endoplasmic reticulum stress

Right ventricular dysfunction parallels left ventricular functional involvement in women with breast cancer experiencing subclinical cardiotoxicity

Abstract Funding Acknowledgements Type of funding sources: None. Background Cancer therapy related cardiac toxicity disease (CRCTD) of the left ventricle (LV)can influence the outcome of oncologic patients. Little is known on CRCTD related right ventricular (RV)dysfunction even though RV involvement has been proven to be a remarkable prognosticator in heart failure. Purpose To analyse parallel changes in LV and RV function occurring during the course of cancer therapy in women affected by breast cancer by using both standard and speckle tracking echocardiography. Methods Fifty Her-2 positive breast cancer women (age = 53.6 ± 11.7 years) underwent sequential cancer therapy protocol including anthracycline (ANT) epirubicine + cyclophosphamide (4 cycles) followed by a total amount of 18 cycles with trastuzumab (TRZ) + paclitaxel. A complete echo-Doppler exam, including LV and RV global longitudinal strain (GLS)as well as RV septal and free wall longitudinal strain (SLS and FWLS respectively) assessment, was performed at baseline, after ANT end and after TRZ completion. Patients with overt heart failure and LV ejection fraction < 50%, coronary artery disease,atrial fibrillation, hemodinamically significant valve disease and inadequate echo were excluded. Overt CRCTD was defined according guidelines and both subclinical LV and RV CRCTD as a LV and RV GLS drop from baseline >15%. Results None of the patients experienced overt CTCRD but 6 patients (14%) showed subclinical LV dysfunction and 33 (66%) had a significant drop of RV longitudinal function.The comparison of standard echo-Doppler exam at baseline and after ANT and TRZ completion did not show significant changes of LV and RV systolic and diastolic parameters. Conversely, a progressive significant reduction of RV GLS (p < 0.002 after TRZ), SLS and FWLS and, with a lower extent, of LV GLS (p < 0.02 after TRZ) was observed after ANT and TRZ completion (Figure). Percentage reduction in RV GLS (DRV GLS) from baseline to ANT end correlated with LV GLS both at EC end (r=-0.40, p = 0.006) and after TRZ completion (r=-0.62, p < 0.0001). Conclusions Detrimental cardiac effects of cancer therapy involve both LV and RV systolic longitudinal function. Progressive RV dysfunction is evident through ANT and TRZ treatment. Early RV dysfunction parallels LV involvement and predicts subsequent LV subclinical dysfunction. A comprehensive LV and RV longitudinal function assessment might better predict the onset of CRCTD in breast cancer patients. Abstract Figure

Evidence for immunomodulation and apoptotic processes induced by cationic polystyrene nanoparticles in the hemocytes of the marine bivalve Mytilus

none8sìPolymeric nanoparticles can reach the marine environment from different sources as weathering of plastic debris and nanowaste. Nevertheless, few data are available on their fate and impact on marine biota. Polystyrene nanoparticles (PS NPs) can be considered as a model for studying the effects of nanoplastics in marine organisms: recent data on amino-modified PS NPs (PS-NH2) toxicity in sea urchin embryos underlined that marine invertebrates can be biological targets of nanoplastics. Cationic PS NPs have been shown to be toxic to mammalian cells, where they can induce apoptotic processes; however, no information is available on their effects and mechanisms of action in the cells of marine organisms. In this work, the effects of 50 nm PS-NH2 were investigated in the hemocytes of the marine bivalve Mytilus galloprovincialis. Hemocytes were exposed to different concentrations (1, 5, 50 μg/ml) of PS-NH2 suspension in ASW. Clear signs of cytoxicity were evident only at the highest concentrations (50 μg/ml). On the other hand, a dose dependent decrease in phagocytic activity and increase in lysozyme activity were observed. PS-NH2 NPs also stimulated increase in extracellular ROS (reactive oxygen species) and NO (nitric oxide) production, with maximal effects at lower concentrations. Moreover, at the highest concentration tested, PS-NH2 NPs induced apoptotic process, as evaluated by Flow cytometry (Annexin V binding and mitochondrial parameters). The results demonstrate that in marine invertebrates the immune function can represent a significant target for PS-NPs. Moreover, in Mytilus hemocytes, PS-NH2 NPs can act through mechanisms similar to those observed in mammalian cells. Further research is necessary on specific mechanisms of toxicity and cellular uptake of nanoplastics in order to assess their impact on marine biota.openCanesi, L; Ciacci, Caterina; Bergami, E; Monopoli, M. P; Dawson, K. A; Papa, Stefano; Canonico, Barbara; Corsi, I.Canesi, L; Ciacci, Caterina; Bergami, E; Monopoli, M. P; Dawson, K. A; Papa, Stefano; Canonico, Barbara; Corsi, I

Neuropsychiatric symptoms and syndromes in a large cohort of newly diagnosed, untreated patients with Alzheimer disease.

Objectives: Neuropsychiatric symptoms are common in patients with Alzheimer disease (AD). Treatment for both AD and psychiatric disturbances may affect the clinical observed pattern and comorbidity. The authors aimed to identify whether particular neuropsychiatric syndromes occur in untreated patients with AD, establish the severity of syndromes, and investigate the relationship between specific neuropsychiatric syndromes and AD disease severity. Design: Cross-sectional, multicenter, clinical study. Participants: A total of 1,015 newly diagnosed, untreated outpatients with AD from five Italian memory clinics were consecutively enrolled in the study from January 2003 to December 2005. Measurements: All patients underwent thorough examination by clinical neurologists/geriatricians, including neuropsychiatric symptom evaluation with the Neuropsychiatric Inventory. Results: Factor analysis revealed five distinct neuropsychiatric syndromes: the apathetic syndrome (as unique syndrome) was the most frequent, followed by affective syndrome (anxiety and depression), psychomotor (agitation, irritability, and aberrant motor behavior), psychotic (delusions and hallucinations), and manic (disinhibition and euphoria) syndromes. More than three quarters of patients with AD presented with one or more of the syndromes (N 790, 77.8%), and more than half exhibited clinically significant severity of symptoms (N 603, 59.4%). With the exception of the affective one, all syndromes showed an increased occurrence with increasing severity of dementia. Conclusions: The authors’ study supports the use of a syndrome approach for neuropsychiatric evaluation in patients with AD. Individual neuropsychiatric symptoms can be reclassified into five distinct psychiatric syndromes. Clinicians should incorporate a thorough psychiatric and neurologic examination of patients with AD and consider therapeutic strategies that focus on psychiatric syndromes, rather than specific individual symptoms

Acellular dermal matrix used in diabetic foot ulcers: Clinical outcomes supported by biochemical and histological analyses

Diabetic foot ulcer (DFU) is a diabetes complication which greatly impacts the patient’s quality of life, often leading to amputation of the affected limb unless there is a timely and adequate management of the patient. DFUs have a high economic impact for the national health system. Data have indeed shown that DFUs are a major cause of hospitalization for patients with diabetes. Based on that, DFUs represent a very important challenge for the national health system. Especially in developed countries diabetic patients are increasing at a very high rate and as expected, also the incidence of DFUs is increasing due to longevity of diabetic patients in the western population. Herein, the surgical approach focused on the targeted use of the acellular dermal matrix has been integrated with biochemical and morphological/histological analyses to obtain evidence-based information on the mechanisms underlying tissue regeneration. In this research report, the clinical results indicated decreased postoperative wound infection levels and a short healing time, with a sound regeneration of tissues. Here we demonstrate that the key biomarkers of wound healing process are activated at gene expression level and also synthesis of collagen I, collagen III and elastin is prompted and modulated within the 28-day period of observation. These analyses were run on five patients treated with Integra® sheet and five treated with the injectable matrix Integra® Flowable, for cavitary lesions. In fact, clinical evaluation of improved healing was, for the first time, supported by biochemical and histological analyses. For these reasons, the present work opens a new scenario in DFUs treatment and follow-up, laying the foundation for a tailored protocol towards complete healing in severe pathological conditions

PREVALENCE OF CANINE DISTEMPER VIRUS IN CANIDS IN CENTRAL ITALY AND FIRST IDENTIFICATION OF ARCTIC LINEAGE IN MARCHE AND UMBRIA REGIONS: A PRELIMINARY STUDY

Canine distemper virus (CDV) is one of the most commonly virus implicated in outbreaks in wild and domestic carnivores. CDV causes severe systemic diseases which normally involves the respiratory, gastrointestinal and nervous systems. To our knowledge the literature about the real incidence of such disease is scarce, particularly in wild animals population. Recently, outbreaks of CDV have been documented in Italian grey wolves (Canis lupus italicus) [1], a least concerned species in IUCN Red List. Therefore, the surveillance of CDV is a priority for the conservation of the wolves and, more generally, for the protection of wild carnivores which are widespread in Central Italy, especially in the National Parks. In total, 215 samples, belonging to 148 canids for CDV presence, were analysed from November 2012 to December 2016 in the laboratory of IZSUM. Of these, 37.2% were dogs, 33% wolves and 29.8% foxes. Animals were collected in 12 different provinces of 6 Regions: Umbria, Marche, Emilia Romagna, Tuscany, Lazio and Apulia. All samples were collected from dead animals which were sent to the Diagnostic Units of Istituto Zoprofilattico Sperimentale Umbria e Marche and subjected to autopsy. The RNA was extracted from organ pools and swabs with a commercial kit, retrotranscripted to cDNA and amplified by the real time PCR with QuantiFast SYBR Green RT PCR kit (Qiagen GmbH, Hilden, Germany) using primers for a fragment of 278bp in CDV nucleoprotein (NP) gene [2]. Samples having a melting temperature (TM) value ±0.5°C versus TM value of positive control were considered positive. Moreover, samples were visualized by UV rays with GelRed TM (Biotium Inc.) after electrophoresis in agarose gels and bands of appropriate sizes were excised, extracted and sequenced. Sequences obtained (n=11) were aligned with NP gene sequences of CDV available in GenBank by MUSCLE. Molecular phylogenetic analysis (MEGA 7.0) was carried out by using Maximum Likelihood method based on the Tamura 3-parameter model. The CDV RNA was identified in 20.3% of the analysed animals. A high positivity rate was identified in dogs with 10.1% of 148 sample tested positive followed by wolves (6.08%) and foxes (5.11%). The Artic Lineage of CDV was identified in 9 out of 11 sequenced samples, in both wild and domestic canids. This strain was identified in 3 different provinces (PU, AP, PG), raising concerns given the vastness of the affected area. Two Onderstepoort strains were also identified. In conclusion, this study shows a wide CDV circulation involving different ecotypes and species in the investigated area. Further studies, based on epidemiological and genetic analysis, will be carried out in order to assess the phylogenetic correlation among the identified strains. This follow up will be important in order to highlight potential risk factors associated with the introduction of this new genotype and to better understand the role played by domestic and wild carnivores interactions in virus spreading. These additional studies should be carried out as soon as possible in order to prevent virus dissemination and to perform ad hoc vaccination campains

Global observational needs and resources for marine biodiversity

Otros autores: Best, B., Brandt, A., Goodwin, K., Iken, A., Marques, A., Miloslavich, P., Ostrowski, M., Turner, W., Achterberg, E., Barry, T., Bigatti, G., Henry, L.A., Ramiro-Sánchez, B., Durán, P., Morato, T., Murray Roberts, J., García-Alegre, A., Cuadrado, M., Murton, B.The diversity of life in the sea is critical to the health of ocean ecosystems that support living resources and therefore essential to the economic, nutritional, recreational, and health needs of billions of people. Yet there is evidence that the biodiversity of many marine habitats is being altered in response to a changing climate and human activity. Understanding this change, and forecasting where changes are likely to occur, requires monitoring of organism diversity, distribution, abundance, and health. It requires a minimum of measurements including productivity and ecosystem function, species composition, allelic diversity, and genetic expression. These observations need to be complemented with metrics of environmental change and socio-economic drivers. However, existing global ocean observing infrastructure and programs often do not explicitly consider observations of marine biodiversity and associated processes. Much effort has focused on physical, chemical and some biogeochemical measurements. Broad partnerships, shared approaches, and best practices are now being organized to implement an integrated observing system that serves information to resource managers and decision-makers, scientists and educators, from local to global scales. This integrated observing system of ocean life is now possible due to recent developments among satellite, airborne, and in situ sensors in conjunction with increases in information system capability and capacity, along with an improved understanding of marine processes represented in new physical, biogeochemical, and biological models

Fas Signalling Promotes Intercellular Communication in T Cells

Cell-to-cell communication is a fundamental process for development and maintenance of multicellular organisms. Diverse mechanisms for the exchange of molecular information between cells have been documented, such as the exchange of membrane fragments (trogocytosis), formation of tunneling nanotubes (TNTs) and release of microvesicles (MVs). In this study we assign to Fas signalling a pivotal role for intercellular communication in CD4+ T cells. Binding of membrane-bound FasL to Fas expressing target cells triggers a well-characterized pro-apoptotic signalling cascade. However, our results, pairing up flow cytometric studies with confocal microscopy data, highlight a new social dimension for Fas/FasL interactions between CD4+ T cells. Indeed, FasL enhances the formation of cell conjugates (8 fold of increase) in an early time-frame of stimulation (30 min), and this phenomenon appears to be a crucial step to prime intercellular communication. Our findings show that this communication mainly proceeds along a cytosolic material exchange (ratio of exchange >10, calculated as ratio of stimulated cells signal divided by that recorded in control cells) via TNTs and MVs release. In particular, inhibition of TNTs genesis by pharmacological agents (Latruculin A and Nocodazole) markedly reduced this exchange (inhibition percentage: >40% and >50% respectively), suggesting a key role for TNTs in CD4+ T cells communication. Although MVs are present in supernatants from PHA-activated T cells, Fas treatment also leads to a significant increase in the amount of released MVs. In fact, the co-culture performed between MVs and untreated cells highlights a higher presence of MVs in the medium (1.4 fold of increase) and a significant MVs uptake (6 fold of increase) by untreated T lymphocytes. We conclude that Fas signalling induces intercellular communication in CD4+ T cells by different mechanisms that seem to start concomitantly with the main pathway (programmed cell death) promoted by FasL

Preventive and curative effect of melatonin on mammary carcinogenesis induced by dimethylbenz[a]anthracene in the female Sprague–Dawley rat

INTRODUCTION: It has been well documented that the pineal hormone, melatonin, which plays a major role in the control of reproduction in mammals, also plays a role in the incidence and growth of breast and mammary cancer. The curative effect of melatonin on the growth of dimethylbenz [a]anthracene-induced (DMBA-induced) mammary adenocarcinoma (ADK) has been previously well documented in the female Sprague–Dawley rat. However, the preventive effect of melatonin in limiting the frequency of cancer initiation has not been well documented. METHODS: The aim of this study was to compare the potency of melatonin to limit the frequency of mammary cancer initiation with its potency to inhibit tumor progression once initiation, at 55 days of age, was achieved. The present study compared the effect of preventive treatment with melatonin (10 mg/kg daily) administered for only 15 days before the administration of DMBA with the effect of long-term (6-month) curative treatment with the same dose of melatonin starting the day after DMBA administration. The rats were followed up for a year after the administration of the DMBA. RESULTS: The results clearly showed almost identical preventive and curative effects of melatonin on the growth of DMBA-induced mammary ADK. Many hypotheses have been proposed to explain the inhibitory effects of melatonin. However, the mechanisms responsible for its strong preventive effect are still a matter of debate. At least, it can be envisaged that the artificial amplification of the intensity of the circadian rhythm of melatonin could markedly reduce the DNA damage provoked by DMBA and therefore the frequency of cancer initiation. CONCLUSION: In view of the present results, obtained in the female Sprague–Dawley rat, it can be envisaged that the long-term inhibition of mammary ADK promotion by a brief, preventive treatment with melatonin could also reduce the risk of breast cancer induced in women by unidentified environmental factors