IGF-I induces upregulation of DDR1 collagen receptor in breast cancer cells by suppressing MIR-199a-5p through the PI3K/AKT pathway.

Discoidin Domain Receptor 1 (DDR1) is a collagen receptor tyrosine-kinase that contributes to epithelial-to-mesenchymal transition and enhances cancer progression. Our previous data indicate that, in breast cancer cells, DDR1 interacts with IGF-1R and positively modulates IGF-1R expression and biological responses, suggesting that the DDR1-IGF-IR cross-talk may play an important role in cancer.In this study, we set out to evaluate whether IGF-I stimulation may affect DDR1 expression. Indeed, in breast cancer cells (MCF-7 and MDA-MB-231) IGF-I induced significant increase of DDR1 protein expression, in a time and dose dependent manner. However, we did not observe parallel changes in DDR1 mRNA. DDR1 upregulation required the activation of the PI3K/AKT pathway while the ERK1/2, the p70/mTOR and the PKC pathways were not involved. Moreover, we observed that DDR1 protein upregulation was induced by translational mechanisms involving miR-199a-5p suppression through PI3K/AKT activation. This effect was confirmed by both IGF-II produced by cancer-associated fibroblasts from human breast cancer and by stable transfection of breast cancer cells with a human IGF-II expression construct. Transfection with a constitutively active form of AKT was sufficient to decrease miR-199a-5p and upregulate DDR1. Accordingly, IGF-I-induced DDR1 upregulation was inhibited by transfection with pre-miR-199a-5p, which also impaired AKT activation and cell migration and proliferation in response to IGF-I.These results demonstrate that, in breast cancer cells, a novel pathway involving AKT/miR-199a-5p/DDR1 plays a role in modulating IGFs biological responses. Therefore, this signaling pathway may represent an important target for breast cancers with over-activation of the IGF-IR axis

Protective effect of procyanidin-rich grape seed extract against Gram-negative virulence factors

Biofilm formation and lipopolysaccharide (LPS) are implicated in the pathogenesis of gastrointestinal (GI) diseases caused by Gram-negative bacteria. Grape seeds, wine industry by-products, have antioxidant and antimicrobial activity. In the present study, the protective effect of procyanidin-rich grape seed extract (prGSE), from unfermented pomace of Vitis vinifera L. cv Bellone, on bacterial LPS-induced oxidative stress and epithelial barrier integrity damage has been studied in a model of Caco-2 cells. The prGSE was characterized at the molecular level using HPLC and NMR. The in vitro activity of prGSE against formation of biofilm of Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli was investigated. In vivo, prGSE activity using infected Galleria mellonella larvae has been evaluated. The results show that the prGSE, if administered with LPS, can significantly reduce the LPS-induced permeability alteration. Moreover, the ability of the extract to prevent Reactive Oxygen Species (ROS) production induced by the LPS treatment of Caco-2 cells was demonstrated. prGSE inhibited the biofilm formation of E. coli and S. Typhimurium. In terms of in vivo activity, an increase in survival of infected G. mellonella larvae after treatment with prGSE was demonstrated. In conclusion, grape seed extracts could be used to reduce GI damage caused by bacterial endotoxin and biofilms of Gram-negative bacteria

Commissioning and improvements of the instrumentation and launch of the scientific exploitation of OARPAF, the Regional Astronomical Observatory of the Antola Park

The OARPAF telescope is an 80-cm-diameter optical telescope installed in the Antola Mount Regional Reserve, in Northern Italy. We present the results of the characterization of the site, as well as developments and interventions that have been implemented, with the goal of exploiting the facility for scientific and educational purposes. During the characterization of the site, an average background brightness of 22.40mAB (B filter) to 21.14mAB (I) per arcsecond squared, and a 1.5″ to 3.0″ seeing, have been measured. An estimate of the magnitude zero points for photometry is also reported. The material under commissioning includes three CCD detectors for which we provide the linearity range, gain, and dark current; a 31-orders échelle spectrograph with R ∼ 8500 to 15,000 and a dispersion of n = 1.39 × 10 − 6 px − 1λ + 1.45 × 10 − 4 nm / px, where λ is expressed in nm. The scientific and outreach potential of the facility is proven in different science cases, such as exoplanetary transits and active galactic nuclei variability. The determination of time delays of gravitationally lensed quasars, the microlensing phenomenon, and the tracking and the study of asteroids are also discussed as prospective science cases

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure < 100 mmHg (n = 1127), estimated glomerular filtration rate < 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

Insulin Resistance: Any Role in the Changing Epidemiology of Thyroid Cancer?

In the past few decades, the incidence of thyroid cancer (TC), namely of its papillary hystotype (PTC), has shown a steady increase worldwide, which has been attributed at least in part to the increasing diagnosis of early stage tumors. However, some evidence suggests that environmental and lifestyle factors can also play a role. Among the potential risk factors involved in the changing epidemiology of TC, particular attention has been drawn to insulin-resistance and related metabolic disorders, such as obesity, type 2 diabetes, and metabolic syndrome, which have been also rapidly increasing worldwide due to widespread dietary and lifestyle changes. In accordance with this possibility, various epidemiological studies have indeed gathered substantial evidence that insulin resistance-related metabolic disorders might be associated with an increased TC risk either through hyperinsulinemia or by affecting other TC risk factors including iodine deficiency, elevated thyroid stimulating hormone, estrogen-dependent signaling, chronic autoimmune thyroiditis, and others. This review summarizes the current literature evaluating the relationship between metabolic disorders characterized by insulin resistance and the risk for TC as well as the possible underlying mechanisms. The potential implications of such association in TC prevention and therapy are discussed

Insulin Receptor Isoform A Modulates Metabolic Reprogramming of Breast Cancer Cells in Response to IGF2 and Insulin Stimulation

Previously published work has demonstrated that overexpression of the insulin receptor isoform A (IR-A) might play a role in cancer progression and metastasis. The IR has a predominant metabolic role in physiology, but the potential role of IR-A in cancer metabolic reprogramming is unknown. We aimed to characterize the metabolic impact of IR-A and its ligand insulin like growth factor 2 (IGF2) in human breast cancer (BC) cells. To establish autocrine IGF2 action, we generated human BC cells MCF7 overexpressing the human IGF2, while we focused on the metabolic effect of IR-A by stably infecting IGF1R-ablated MCF7 (MCF7IGF1R-ve) cells with a human IR-A cDNA. We then evaluated the expression of key metabolism related molecules and measured real-time extracellular acidification rates and oxygen consumption rates using the Seahorse technology. MCF7/IGF2 cells showed increased proliferation and invasion associated with aerobic glycolysis and mitochondrial biogenesis and activity. In MCF7IGF1R-ve/IR-A cells insulin and IGF2 stimulated similar metabolic changes and were equipotent in eliciting proliferative responses, while IGF2 more potently induced invasion. The combined treatment with the glycolysis inhibitor 2-deoxyglucose (2DG) and the mitochondrial inhibitor metformin blocked cell invasion and colony formation with additive effects. Overall, these results indicate that IGF2 and IR-A overexpression may contribute to BC metabolic reprogramming

Discoidin domain receptor 1 modulates insulin receptor signaling and biological responses in breast cancer cells.

The fetal isoform A of the insulin receptor (IR-A) is frequently overexpressed in a variety of malignancies including breast cancer. IR overexpression has a recognized role in cancer progression and resistance to anticancer therapies. In particular, IR-A has a peculiar mitogenic potential and is activated not only by insulin but also by IGF-2. Previously, we identified discoidin domain receptor 1 (DDR1) as a new IR-A interacting protein. DDR1, a non-integrin collagen tyrosine kinase receptor, is overexpressed in several malignancies and plays a role in cancer progression and metastasis.We now evaluated whether DDR1 is able to exert a role in breast cancer biology by functionally cross-talking with IR. In MCF-7 human breast cancer cells, IR and DDR1 co-immunoprecipitated and co-localized after insulin or IGF-2 stimulation. In a panel of breast cancer cells, DDR1 knockdown by specific siRNAs markedly inhibited IR downstream signaling as well as proliferation, migration and colony formation in response to insulin and IGF-2. These effects were accompanied by reduction of IR protein and mRNA expression, which involved both transcriptional and post-transcriptional effects. DDR1 overexpression elicited opposite effects. Bioinformatics analysis of public domain databases showed that IR and DDR1 co-expression significantly correlates with several clinically relevant histopathological and molecular features of human breast carcinomas.These findings demonstrate that, in human breast cancer cells, DDR1 regulates IR expression and ligand dependent biological actions. This novel functional crosstalk is likely clinically relevant and may become a new molecular target in breast cancer

Short-chain fatty acids promote the effect of environmental signals on the gut microbiome and metabolome in mice

Gut microorganisms and the products of their metabolism thoroughly affect host brain development, function and behavior. Since alterations of brain plasticity and cognition have been demonstrated upon motor, sensorial and social enrichment of the housing conditions, we hypothesized that gut microbiota and metabolome could be altered by environmental stimuli, providing part of the missing link among environmental signals and brain effects. In this preliminary study, metagenomic and metabolomic analyses of mice housed in different environmental conditions, standard and enriched, identify environment-specific microbial communities and metabolic profiles. We show that mice housed in an enriched environment have distinctive microbiota composition with a reduction in gut bacterial richness and biodiversity and are characterized by a metabolomic fingerprint with the increase of formate and acetate and the decrease of bile salts. We demonstrate that mice treated with a mixture of formate and acetate recapitulate some of the brain plasticity effects modulated by environmental enrichment, such as hippocampal neurogenesis, neurotrophin production, short-term plasticity and cognitive behaviors, that can be further exploited to decipher the mechanisms involved in experience-dependent brain plasticity

Protective Effect of Procyanidin-Rich Grape Seed Extract against Gram-Negative Virulence Factors

Biofilm formation and lipopolysaccharide (LPS) are implicated in the pathogenesis of gastrointestinal (GI) diseases caused by Gram-negative bacteria. Grape seeds, wine industry by-products, have antioxidant and antimicrobial activity. In the present study, the protective effect of procyanidin-rich grape seed extract (prGSE), from unfermented pomace of Vitis vinifera L. cv Bellone, on bacterial LPS-induced oxidative stress and epithelial barrier integrity damage has been studied in a model of Caco-2 cells. The prGSE was characterized at the molecular level using HPLC and NMR. The in vitro activity of prGSE against formation of biofilm of Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli was investigated. In vivo, prGSE activity using infected Galleria mellonella larvae has been evaluated. The results show that the prGSE, if administered with LPS, can significantly reduce the LPS-induced permeability alteration. Moreover, the ability of the extract to prevent Reactive Oxygen Species (ROS) production induced by the LPS treatment of Caco-2 cells was demonstrated. prGSE inhibited the biofilm formation of E. coli and S. Typhimurium. In terms of in vivo activity, an increase in survival of infected G. mellonella larvae after treatment with prGSE was demonstrated. In conclusion, grape seed extracts could be used to reduce GI damage caused by bacterial endotoxin and biofilms of Gram-negative bacteria

Valutazione in vitro dell'attività antibatterica e antitumorale della frazione peptidica estratta dall'emolinfa di Hermetia illucens

La resistenza agli antibiotici rappresenta una delle emergenze sociali e sanitarie più urgenti a livello globale insieme alla resistenza alle terapie contro il cancro: l'uso di agenti chemioterapici convenzionali e l'uso indiscriminato e prolungato di antibiotici ha contribuito rispettivamente allo sviluppo di cellule tumorali resistenti e microrganismi (virus, batteri e funghi) resistenti alle terapie convenzionali. Nella ricerca di nuovi farmaci sicuri ed efficaci, è stata recentemente prestata maggiore attenzione a una classe di composti biologicamente attivi: i peptidi antimicrobici (AMP), piccole proteine bioattive, naturalmente prodotte da tutti gli organismi viventi come componenti dell’immunità innata. Gli AMP possono avere numerosi vantaggi rispetto agli antibiotici convenzionali, quali bassi livelli di resistenza, attività ad ampio spettro con tossicità ospite minima, effetti sinergici e rapida uccisione del microrganismo. Molti studi hanno anche dimostrato che alcuni AMP presentano attività citotossica contro le cellule tumorali e sono definiti come peptidi antitumorali (ACP). Le interazioni fra le membrane dei microrganismi e gli AMP dipendono dalle forze elettrostatiche fra gli AMP carichi positivamente e la membrana dei microorganismi carica negativamente per la presenza di acido teicoico sulla parete di batteri Gram positivi e del lipolisaccaride sulla parete esterna dei batteri Gram negativi, con un meccanismo di azione che può essere membranolitico e non membranolitico. Similmente ai batteri, le cellule tumorali hanno una carica netta negativa dovuta all’elevata espressione di molecole anioniche, quali fosfoatidilserina e mucine O-glicosilate, sul lato esterno della membrana; questo permette interazioni elettrostatiche tra gli ACP e la superficie di molte cellule tumorali. Inoltre, gli ACP possono destabilizzare più facilmente la membrana delle cellule tumorali data la sua maggiore fluidità e la maggiore presenza, rispetto alle cellule sane, di microvilli sulla superficie cellulare e possono, una volta penetrati all’interno della cellula, interrompere l'integrità delle membrane mitocondriali cariche negativamente, con conseguente rilascio di potenti proteine pro-apoptotiche. Gli AMP sono prodotti da tutti gli organismi, ma gli insetti sono tra le fonti più ricche e innovative. Lo scopo della nostra ricerca è quello di individuare principi farmacologicamente attivi di origine naturale a partire dall'insetto Hermetia illucens (Diptera, Stratiomyidae), da utilizzare per lo sviluppo di farmaci antimicrobici e antitumorali alternativi o a sostegno di terapie convenzionali già in uso. La componente peptidica è stata isolata dall'emolinfa di larve immunizzate (con batteri Gram-positivi o Gram-negativi) e non immunizzate e analizzato per valutare l’attività antimicrobica e anticancro. Le proprietà antimicrobiche sono state analizzate mediante saggi di diffusione in agar e microdiluizioni scalari contro batteri Gram-positivi e Gram-negativi, mentre l'effetto antitumorale è stato valutato mediante saggio MTT in diverse linee cellulari tumorali