The addition of long-acting beta2-agonists (LABAs) to corticosteroids improves asthma control. Cigarette smoke exposure, increasing oxidative stress, may negatively affect corticosteroid responses. ...The anti-inflammatory effects of formoterol (FO) and fluticasone propionate (FP) in human bronchial epithelial cells exposed to cigarette smoke extracts (CSE) are unknown.
This study explored whether FP, alone and in combination with FO, in human bronchial epithelial cellline (16-HBE) and primary bronchial epithelial cells (NHBE), counteracted some CSE-mediated effects and in particular some of the molecular mechanisms of corticosteroid resistance.
16-HBE and NHBE were stimulated with CSE, FP and FO alone or combined. HDAC3 and HDAC2 activity, nuclear translocation of GR and NF-κB, pERK1/2/tERK1/2 ratio, IL-8, TNF-α, IL-1β mRNA expression, and mitochondrial ROS were evaluated. Actin reorganization in neutrophils was assessed by fluorescence microscopy using the phalloidin method.
In 16-HBE, CSE decreased expression/activity of HDAC3, activity of HDAC2, nuclear translocation of GR and increased nuclear NF-κB expression, pERK 1/2/tERK1/2 ratio, and mRNA expression of inflammatory cytokines. In NHBE, CSE increased mRNA expression of inflammatory cytokines and supernatants from CSE exposed NHBE increased actin reorganization in neutrophils. FP combined with FO reverted all these phenomena in CSE stimulated 16-HBE cells as well as in NHBE cells.
The present study provides compelling evidences that FP combined with FO may contribute to revert some processes related to steroid resistance induced by oxidative stress due to cigarette smoke exposure increasing the anti-inflammatory effects of FP.
•Cigarette smoke negatively affects corticosteroid responses.•Airway epithelium is a primary target of the anti-inflammatory actions of glucocorticosteroids.•Formoterol enhances the effects of Fluticasone Propionate and in this way counteracts some events related to steroid resistance induced by cigarette smoke.
Background
Alterations in the nasal epithelial barrier homeostasis and increased interleukin 33 (IL‐33) expression contribute to the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP).
...Aims
As Notch‐1 signaling is crucial in repair processes of mucosa, the current study assessed Notch‐1/Jagged‐1 signaling and IL‐33 in the epithelium of nasal polyps biopsies from allergic (A‐CRSwNP;
n = 9) and not allergic (NA‐CRSwNP;
n = 9) subjects by immunohistochemistry. We also assessed, in a model of nasal epithelial cells, the effects of stimulation of Notch‐1 with Jagged‐1 on the expression of IL‐33 (by flow cytometry, immunofluorescence, and immunocytochemistry), Jagged‐1 (by flow cytometry), and p‐CREB transcription factor (by western blot analysis).
Results
Ex vivo (a) in normal epithelium, the expression of Notch‐1 and IL‐33 were higher in NA‐CRSwNP than in A‐CRSwNP; (b) in metaplastic epithelium, the expression of Notch‐1, Jagged‐1, and IL‐33 were higher in NA‐CRSwNP than in A‐CRSwNP; (c) in hyperplastic epithelium, the expression of Notch‐1, Jagged‐1, and IL‐33 were higher in A‐CRSwNP than in NA‐CRSwNP; and (d) in basal epithelial cells, no differences were observed in the expression of Jagged‐1, IL‐33, and Notch‐1. The expression of Notch‐1 significantly correlated with the expression of IL‐33. In vitro, stimulation of Notch‐1 with Jagged‐1 induced the expression of (a) Jagged‐1; (b) IL‐33; and (c) p‐CREB transcription factor. The inhibitor of Notch‐1, DAPT, reduced all the effects of Jagged‐1 on nasal epithelial cells.
Conclusions
The data herein provided support, for the first time, a putative role of Notch‐1/Jagged‐1 signaling in the overexpression of IL‐33 in the epithelium of nasal polyps from patients with CRSwNP.
The current study assessed for the first time Notch‐1, Jagged‐1, and interleukin 33 expression in the epithelium of CRSwNP from allergic and not allergic exsmoker subjects. The data demonstrated a role of Notch‐1/Jagged‐1 signaling in a positive feedback upregulating Jagged‐1 expression and in promoting proinflammatory responses inducing p‐CREB and IL‐33 expression in nasal epithelial cells.
The sensitivity of gravitational wave interferometric detectors is ultimately limited by the quantum noise, which arises from the quantum nature of light and it is driven by vacuum fluctuations of ...the optical field entering from the dark port of the interferometer. One way to improve the sensitivity of gravitational wave interferometers is to inject squeezed vacuum into the dark port. This has been already demonstrated for the main gravitational wave detectors (GEO, Advanced LIGO and Advanced VIRGO). We are studying tricks to produce a "frequency- dependent squeezing": a standard method is to filter the squeezed optical field with one or more optical cavities (300 m long cavities). An alternative method using a pair of squeezed EPR (Einstein-Podolsky-Rosen) entangled beams to produce frequency-dependent squeezing by a non-degenerate OPO (Optical Parametric Oscillator) will be discussed in this paper. This method promises to achieve a frequency-dependent optimization of the injected squeezed light fields without the need for an external filter cavity.
Gram-negative bacteria are an uncommon etiology of spontaneous community-acquired adult meningitis and meningoencephalitis. Escherichia coli is a Gram-negative bacterium that is normally present in ...the intestinal microbial pool. Some Escherichia coli strains can cause diseases in humans and animals, with both intestinal and extraintestinal manifestations (extraintestinal pathogenic Escherichia coli) such as urinary tract infections, bacteremia with sepsis, and, more rarely, meningitis. Meningitis continues to be an important cause of mortality throughout the world, despite progress in antimicrobial chemotherapy and supportive therapy. The mortality rate fluctuates between 15% and 40%, and about 50% of the survivors report neurological sequelae. The majority of Escherichia coli meningitis cases develop as a result of hematogenous spread, with higher degrees of bacteremia also being related to worse prognosis. Cases presenting with impaired consciousness (that is, coma) are also reported to have poorer outcomes.
We describe the case of a 48-year-old caucasian woman with meningoencephalitis, with a marked alteration of consciousness on admission, and septic shock secondary to pyelonephritis caused by Escherichia coli, treated with targeted antimicrobial therapy and immunoglobulin-M-enriched immunoglobulin (Pentaglobin) preparation as adjuvant therapy.
Despite the dramatic presentation of the patient on admission, the conflicting data on the use of immunoglobulins in septic shock, and the lack of evidence regarding their use in adult Escherichia coli meningoencephalitis, we obtained a remarkable improvement of her clinical condition, accompanied by partial resolution of her neurological deficits.
•Amperometric sensor for detection of H2O2 released by living cells (human THP-1 macrophages) was developed.•H2O2 diffuses through cell membrane and can be checked in the extracellular ...space.•Macrophages highly provide Reactive Oxygen Species generation in inflamed tissues.•Macrophages were exposed to pro- and anti-oxidant treatments and H2O2 release was measured in conditioned medium.•Results of the electrochemical sensor agree with those obtained by flow cytometry using the same cells.
Increased oxidative burden contributes to the pathogenesis of most inflammatory diseases and is associated with aging and chronic inflammation. Macrophages contribute to the generation of reactive oxygen species (ROS) within inflamed tissues. Currently, ROS generation is measured using fluorescent probes and colorimetric/fluorimetric biochemical assays. Hydrogen peroxide (H2O2) diffuses through the cell membrane and can be monitored in the extracellular space. Herein, we present a sensor for H2O2 detection released by cells in culture supernatants. H2O2 sensing performance was evaluated using chronoamperometric detection. A sensitivity of 0.0641 μA μM−1 cm−2 with a limit of detection of 6.55 μM and excellent selectivity against many interferents was found. H2O2 release was also measured in conditioned medium from human THP-1 macrophages exposed to pro-oxidant and anti-oxidant treatments. The results were compared with those obtained by flow cytometry using the same cells stained with carboxy-H2DCFDA and MitoSOX Red, which detect intracellular ROS and mitochondrial superoxide, respectively. The addition of pro-oxidants lipopolysaccharide (LPS) and nigericin resulted in a significant increase in the cathodic current due to the H2O2 reduction, indicating an increased release of H2O2. The addition of 17-oxo-DHA, which inhibits LPS- and nigericin-dependent responses, decreased the LPS- and nigericin-induced release of H2O2. All the results obtained with the sensor were consistent with those obtained using flow cytometry. The operation of the sensor directly in the cell culture growth medium had no impact on cell viability. The sensor is highly sensitive, fast, and cost effective, and it can potentially be used for real time monitoring of oxidative stress.
In this paper, we summarize the present state-of-the-art on the proof-of-principle experiment of frequency-dependent squeezing implemented through EPR entanglement for Virgo gravitational-wave ...detector and we introduce Virgo subsystem proposal for frequency-dependent squeezing, obtained with a compact apparatus and without the costs required by the infrastructure for the filter cavity.
•Cigarette smoke is considered a risk factor for lung and colorectal cancer.•Cigarette smoke induces epithelial mesenchymal transition features in Caco2.•Cigarette smoke increases the invasive ...potential of Caco2.
Cigarette smoke is considered a risk factor for lung and colorectal cancer. A convincing link between epithelial-to-mesenchymal transition (EMT) with colorectal cancer progression and therapeutic resistance has emerged. Deregulated expression of E-Cadherin and Claudin-1 and increased miR-21 expression and invasiveness represent hallmarks of EMT. The effects of cigarette smoke exposure on EMT in colorectal adenocarcinoma cells are largely unknown.
The aim of the study is to evaluate the effect of cigarette smoke extract (CSE) on miR-21, Claudin-1 and E-Cadherin, molecules associated to EMT in colorectal cancer cells.
A human colorectal adenocarcinoma cell line (Caco-2) was treated with CSE at different concentration (5% and 10%) and for different time points (3 h and 24 h). Metabolic activity (by MTS assay), cell necrosis/cell apoptosis (evaluating Propidium Iodide/Annexin V expression by flow cytometry), miR-21, Claudin-1 and E-Cadherin gene expression were evaluated by Real time PCR. Cell permeability, actin polymerization and cancer cell migration was assessed by Trans-Epitelial Electrical Resistance (TEER), Phalloidin expression and matrigel system, respectively.
CSE at all the tested concentrations and at all time points reduced cell necrosis. CSE at 10% increased miR-21 and reduced the metabolic activity, cell necrosis, Claudin-1 and E-cadherin mRNA at 3 h. Cell permeability, actin polymerization and cancer cell migration were all increased upon CSE exposure.
These results showed that CSE increasing miR-21, Claudin-1 and E-Cadherin and enhancing the aggressiveness of cancer cells, may concur to colorectal cancer progression.
Third-generation gravitational wave observatories will extend the lower frequency limit of the observation band toward 2 Hz, where new sources of gravitational waves, in particular intermediate-mass ...black holes (IMBH), will be detected. In this frequency region, seismic noise will play an important role, mainly through the so-called Newtonian noise, i.e., the gravity-mediated coupling between ground motion and test mass displacements. The signal lifetime of such sources in the detector is of the order of tens of seconds. In order to determine whether a candidate site to host the Einstein Telescope observatory is particularly suitable to observe such sources, it is necessary to estimate the probability distributions that, in the characteristic time scale of the signal, the sensitivity of the detector is not perturbed by Newtonian noise. In this paper, a first analysis is presented, focused on the Sos Enattos site (Sardinia, Italy), a candidate to host the Einstein Telescope. Starting from a long data set of seismic noise, this distribution is evaluated considering both the presently designed triangular ET configuration and also the classical ”L” configuration.
High concentrations of H2O2, indicative of increased oxidative stress in the lung, are observed in the exhaled breath of individuals affected by different respiratory diseases. Therefore, measuring ...H2O2 in exhaled breath represents a promising and non-invasive approach for monitoring the onset and progression of these diseases. Herein, we have developed an innovative, inexpensive, and easy-to-use device for the measurement of H2O2 in exhaled breath. The device is based on a silver layer covered with an electrodeposited thin film of chitosan, that ensures the wettability of the sensor in a humid atmosphere. The s-ensor was calibrated in the aerosol phase using both phosphate buffer solution and cell culture medium. In the buffer, a sensitivity of 0.110 ± 0.0042 μA μM−1 cm−2 (RSD: 4%) and a limit of detection of 30 μM were calculated, while in the cell culture medium, a sensitivity of 0.098 ± 0.0022 μA μM−1 cm−2 (RSD 2%) and a limit of detection of 40 μM were obtained. High selectivity to different interfering species was also verified. The sensor was further tested versus an aerosol phase obtained by nebulizing the culture medium derived from human bronchial epithelial cells that had been exposed to pro-oxidant and antioxidant treatments. The results were comparable with those obtained using the conventional cytofluorimetric method. Finally, sensor was tested in real exhaled breath samples and even after undergoing physical deformations. Data herein presented support that in future applications this device can be integrated into face masks allowing for easy breath monitoring.
•H2O2 is a good biomarker of oxidative stress related to respiratory diseases.•An innovative, cheap and non-invasive device for monitoring H2O2 was proposed.•An electrochemical sensor, easily integrable inside a face mask, was developed.•A complete device was obtained from the silver layer of waste compact discs.•H2O2 was quantified in the nebulized phase of the medium of cellular growth.
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