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Aims: The angiotensin receptor neprilysin inhibitor (ARNI) sacubitril/valsartan reduces mortality and hospitalizations in patients with heart failure and reduced ejection fraction (HFrEF). Favourable effects on haemodynamic and functional parameters have been observed in patients with HFrEF undergoing ARNI therapy, using standard transthoracic echocardiography. Global longitudinal strain (GLS) assessment uses a semi-automatic procedure to provide a reliable and repeatable method that improves the detection of early changes of contractile function. We aimed to assess the effects of ARNI on GLS and myocardial mechanics in patients with HFrEF., Methods and Results: Thirty patients with New York Heart Association class II-III HFrEF were treated with ARNI and monitored using standard echocardiographic examination and GLS measurements at baseline, 3 months, and 6 months. ARNI therapy resulted in a significant reduction of ventricular volumes and a significant increase in left ventricular ejection fraction at 6 months but not 3 months by standard transthoracic echocardiography (left ventricular ejection fraction from 28 ± 8% at baseline to 34 ± 12% at 6 months, P < 0.001). Non-significant differences in the size of the left atrium, right ventricular function, and pulmonary pressures were found at 6 months. By using GLS, there was a progressive improvement of all strain parameters by 3 months. The improvement showed a progressive trend over time and maintained significance at 6 months: GLS 4ch -7.2 ± 4.8% at baseline vs. -7.5 ± 3.9% at 3 months (P = 0.025) and - 9.2 ± 5.2% at 6 months (P = 0.0001); AVG GLS -6.9 ± 4.3 at baseline vs. -7.9 ± 4.2 at 3 months (P = 0.04) and - 8.8 ± 4.4 at 6 months (P = 0.035); GLS endo 8.2 ± 4.8 at baseline vs. -9.0 ± 4.8 at 3 months (P = 0.05) and - 10.1 ± 5.1 at 6 months (P = 0.001)., Conclusions: Sacubitril/valsartan induces an early benefit on left ventricular remodelling, which is captured by myocardial strain and not by standard echocardiography. Strain method represents a practical tool to assess early and minimal variations of left ventricular systolic function., (© 2020 The Authors. ESC Heart Failure published by John Wiley & Sons Ltd on behalf of the European Society of Cardiology.)

Background: Using antibodies against intracellular phosphoproteins, flow cytometry can be used to monitor simultaneously multiple signaling pathways. Here, we tested a recently released procedure to analyze phosphorylation events in human monocytes upon different types of stimulation., Methods: Whole blood was treated by lipopolysaccharide (LPS) or granulocyte-macrophage colony-stimulating factor (GM-CSF), then cells were labeled by antibodies recognizing cell surface and cytosolic proteins. Human monocytes were identified by a CD14 - CD45 staining and three phosphorylated proteins such as AKT, ERK-1/2, and STAT5, were simultaneously detected by multicolor phosphoflow analysis., Results: By this rapid method, we are able to detect directly from a blood sample several signaling events in human monocytes where LPS stimulation induces preferentially ERK-1/2 phosphorylation where as GM-CSF stimulation induces STAT5 phosphorylation., Conclusions: This procedure provides a simultaneous measurement of multiple activated signaling molecules using a simplified and rapid protocol. © 2015 International Clinical Cytometry Society., (© 2015 Clinical Cytometry Society.)

Drug discovery often requires the screening of compound libraries on tissue cultured cells. Some major targets in drug discovery belong to signal transduction pathways, and PerFix EXPOSE* allows easy flow cytometry phospho assays. We thus investigated the possibility to further simplify and automate this assay, to allow the direct screening of drugs targeting signaling pathways. We show here the sensitivity of this fully automated assay on human growth hormone (hGH)-driven JAK/STAT5-activated IM-9 cells, and we discuss the throughput of this system, which is compatible with medium-throughput drug screening. Because the kit works directly on whole blood samples, ex-vivo assays are also possible with this approach, which could allow for the screening of drugs under more physiological conditions., (© 2014 Society for Laboratory Automation and Screening.)

Purpose: Oxidative damage and inflammation are expected to be involved in age-related functional decline of lachrymal gland, that induces lachrymal dysfunction; this resulting in dry eye disease. Therefore, we investigated the potential antioxidant effect of 0.2% xanthan gum (XNT) in human corneal epithelial cells (HCE), in comparison with other widely used tear substitute polymers, such as 0.2% hydroxyethylcellulose (HEC), 0.2% hyaluronic acid (HA) and 0.5% carboxymethylcellulose (CMC)., Methods: Subconfluent (80%) HCE (Human Corneal Epithelial) cultures were treated with the different polysaccharides at the above reported concentrations. The effect of every polymer was investigated with and without 0.5 mM H2O2 In detail, hydrogen peroxide was added 1 hour after the addition of polysaccharides. Twelve hours later, reactive oxygen species (ROS) production (dichlorofluorescein diacetate spectrofluorimetric test) was assessed and their values were normalized versus protein content. Morphological analysis was performed by optical microscopy., Results: No morphological differences in HCE compared to control cells (CTRL, cells treated with the buffer used for polymer solubilization) were observed in any of the tested polymers, whereas, in the presence of 0.5 mM H2O2 HCE clearly showed signs of cytotoxicity. Polymers did protect cultures from oxidative stress with XNT>HA = HEC>CMC, as evidenced by microscopic analysis. These results were confirmed from ROS measurements, which showed XNT as the only polysaccharide to restore the levels of ROS comparable to CTRL, in presence of H2O2., Conclusions: 0.2% xanthan gum was able to protect HCE by oxidative stress, bringing the ROS level down to CTRL values. Considering that in dry eye syndrome oxidative stress sustains inflammation and apoptotic cell death, the use of xanthan gum in ophthalmic preparations could be beneficial.