A Novel Hybrid Platform for Live/Dead Bacteria Accurate Sorting by On-Chip DEP Device.

According to the World Health Organization (WHO) forecasts, Antimicrobial Resistance (AMR) will be the leading cause of death worldwide in the next decades. To prevent this phenomenon, rapid Antimicrobial Susceptibility Testing (AST) techniques are required to drive the selection of the most suitable antibiotic and its dosage. In this context, we propose an on-chip platform, based on a micromixer and a microfluidic channel, combined with a pattern of engineered electrodes to exploit the di-electrophoresis (DEP) effect. The role of the micromixer is to ensure the proper interaction of the antibiotic with the bacteria over a long time (≈1 h), and the DEP-based microfluidic channel enables the efficient sorting of live from dead bacteria. A sorting efficiency of more than 98%, with low power consumption (Vpp = 1 V) and time response of 5 s, within a chip footprint of ≈86 mm2, has been calculated, which makes the proposed system very attractive and innovative for efficient and rapid monitoring of the antimicrobial susceptibility at the single-bacterium level in next-generation medicine.

Tunable narrow band add-drop filter design based on apodized long period waveguide grating assisted co-directional coupler.

Tunable add/drop filter based optical interconnects are an integral part of data centers as well as optical communications. Although add/drop filters based on ring resonators and waveguide Bragg gratings are well developed, long period waveguide grating (LPWG) based add/drop filters have little been investigated so far. In this article, we propose an apodized LPWG assisted co-directional coupler for narrow band add/drop filtering by combining silicon (Si) waveguide with titanium dioxide (TiO2) waveguide geometry. The proposed structure has been analyzed by combining the finite element method (FEM) and transfer matrix method (TMM), showing a good side lobe suppression ratio (SLSR) equal to 25.7 dB and an insertion loss of 0.6 dB. Owing to the high group index difference of Si and TiO2 waveguides, a narrow band response of 1.4 nm has been achieved with 800µm long LPWG. The opposite thermo-optic coefficients of Si and TiO2 ensures a good thermal tunability of the central wavelength. Considering a thin metallic heater of titanium nitride (TiN) the thermal tuning efficiency is found to be 0.07 nm/mW. Further, two LPWGs have been cascaded to realize a tunable dual channel filter with a minimum channel spacing of 185 GHz and a channel crosstalk better than 20 dB, showing its potential application towards dense wavelength division multiplexing.

On the Use of Mechanistic Soil-Plant Uptake Models: A Comprehensive Experimental and Numerical Analysis on the Translocation of Carbamazepine in Green Pea Plants.

Food contamination is a major worldwide risk for human health. Dynamic plant uptake of pollutants from contaminated environments is the preferred pathway into the human and animal food chain. Mechanistic models represent a fundamental tool for risk assessment and the development of mitigation strategies. However, difficulty in obtaining comprehensive observations in the soil-plant continuum hinders their calibration, undermining their generalizability and raising doubts about their widespread applicability. To address these issues, a Bayesian probabilistic framework is used, for the first time, to calibrate and assess the predictive uncertainty of a mechanistic soil-plant model against comprehensive observations from an experiment on the translocation of carbamazepine in green pea plants. Results demonstrate that the model can reproduce the dynamics of water flow and solute reactive transport in the soil-plant domain accurately and with limited uncertainty. The role of different physicochemical processes in bioaccumulation of carbamazepine in fruits is investigated through Global Sensitivity Analysis, which shows how soil hydraulic properties and soil solute sorption regulate transpiration streams and bioavailability of carbamazepine. Overall, the analysis demonstrates the usefulness of mechanistic models and proposes a comprehensive numerical framework for their assessment and use.

Measured radiation effects on InGaAsP/InP ring resonators for space applications.

Photonic ring resonators can be considered building blocks of new concept satellite payloads for implementing several functions, such as filtering and sensing. In particular, the use of a high Q-factor ring resonator as sensing element into a Resonant Micro Optic Gyroscope (RMOG), provides a remarkable improvement of the performance with respect to the competitive technologies. To qualify a ring resonator for Space applications, the radiation effects on it in the Space must be carefully evaluated. Here, we investigate the effects of gamma radiation on a high Q InGaAsP/InP ring resonator, for the first time, to our knowledge. The ring resonator under study has a footprint of about 530 mm2 and it is based on a InGaAsP/InP rib waveguide, with a width of 2 µm and a thickness of 0.3 µm, formed on a 0.7 µm thick slab layer on an InP substrate 625 µm thick. For a total dose of about 320 krad Co60 gamma irradiation, a mean variation of about 13% and 4% was measured for Q and extinction ratio (ER), respectively, with respect to the values before irradiation (Q = 1.36 × 106, ER = 6.24 dB). Furthermore, the resonance peak red-shifts with a linear behaviour was observed increasing the total dose of the absorbed radiation, with a maximum resonance detuning of about 810 pm. These non-significant effects of a quite high gamma radiation dose confirm the potential of high-Q InP-based ring resonators into Space systems or subsystems.

Design of an ultra-compact graphene-based integrated microphotonic tunable delay line.

The design of a continuously tunable optical delay line based on a compact graphene-based silicon Bragg grating is reported. High performance, in terms of electro-optical switching time (tswitch < 8 ns), delay range (Δτ = 200 ps), and figure of merit FOM = Δτ/A = 1.54x105 ps/mm2, has been achieved with an ultra-compact device footprint (A ~1.3 x 10-3 mm2), so improving the state-of-the-art of integrated optical delay lines. A continuous and complete tunability of the delay time can be achieved with a very low delay loss ( = 0.03 dB/ps) and a weak power consumption ( = 0.05 mW/ps). A flat bandwidth B = 1.19 GHz has been calculated by exploiting the slow-light effect in the device. This performance makes the proposed optical delay line suitable for several applications in Microwave Photonics (MWP), such as beamsteering/beamforming, for which large delay range, flat and wide bandwidth and small volume are required.

Long-term home noninvasive mechanical ventilation increases systemic inflammatory response in chronic obstructive pulmonary disease: a prospective observational study.

BACKGROUND: Long-term home noninvasive mechanical ventilation (NIV) is beneficial in COPD but its impact on inflammation is unknown. We assessed the hypothesis that NIV modulates systemic and pulmonary inflammatory biomarkers in stable COPD. METHODS: Among 610 patients referred for NIV, we shortlisted those undergoing NIV versus oxygen therapy alone, excluding subjects with comorbidities or non-COPD conditions. Sputum and blood samples were collected after 3 months of clinical stability and analyzed for levels of human neutrophil peptides (HNP), interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF-alpha). Patients underwent a two-year follow-up. Unadjusted, propensity-matched, and pH-stratified analyses were performed. RESULTS: Ninety-three patients were included (48 NIV, 45 oxygen), with analogous baseline features. Sputum analysis showed similar HNP, IL-6, IL-10, and TNF-alpha levels (P > 0.5). Conversely, NIV group exhibited higher HNP and IL-6 systemic levels (P < 0.001) and lower IL-10 concentrations (P < 0.001). Subjects undergoing NIV had a significant reduction of rehospitalizations during follow-up compared to oxygen group (P = 0.005). These findings were confirmed after propensity matching and pH stratification. CONCLUSIONS: These findings challenge prior paradigms based on the assumption that pulmonary inflammation is per se detrimental. NIV beneficial impact on lung mechanics may overcome the potential unfavorable effects of an increased inflammatory state.

Effect of particle density on microplastics transport in artificial and natural porous media.

The occurrence and persistence of microplastics (MPs) in natural environments are of increasing concern. Along with this, the transport of MPs in sediments has been investigated mainly focusing on the effect of plastic size and shape, media size effect, and solution chemistry. Yet, the influence of particle density is only partially understood. Therefore, column experiments on the transport of variably buoyant MPs in saturated natural sediments and glass beads were conducted, and transport parameters were quantified using a two-site kinetic transport model with a depth-dependent blocking function (the amount of retained MPs does not decrease at a constant rate with increasing depth, the majority of MPs were retained near the column inlet). Neutral, sinking, and buoyant MPs within the same size range were selected, with stable water isotope applied as conservative tracer to explore water and MP movement in the tested sediments. The results showed that 95.5 ± 1.4% of sinking MPs remained in columns packed with gravel, followed by buoyant and neutral MPs, thus indicating that particle density does affect MP mobility. Similar recovered amounts of MPs were found in columns packed with glass beads, indicating that tested sediment types do not affect the deposition behavior of MPs. The breakthrough curves of MPs were accurately described by the selected model. However, the simulated retention profiles overestimated the observed MP amount in layers closest to the column inlet. The coupled experimental and modeled results suggest an enhanced retention of sinking MPs, while neutrally and buoyant MPs exhibit a higher mobility in comparison. Thus, neutral or buoyant MPs can potentially pose a higher contamination risk to subsurface porous media environments compared to sinking MPs. Discrepancies between observed and simulated retention profiles indicate that future model development is needed for advancing the MP deposition as affected by particle density.

Experimental and simulated microplastics transport in saturated natural sediments: Impact of grain size and particle size.

Microplastics (MPs) present in terrestrial environments show potential leaching risk to deeper soil layers and aquifer systems, which threaten soil health and drinking water supply. However, little is known about the environmental fate of MPs in natural sediments. To examine the MPs transport mechanisms in natural sediments, column experiments were conducted using different natural sediments and MPs (10-150 µm) with conservative tracer. Particle breakthrough curves (BTCs) and retention profiles (RPs) were numerically interpreted in HYDRUS-1D using three different models to identify the most plausible deposition mechanism of MPs. Results show that the retention efficiency for a given particle size increased with decreasing grain size, and RPs exacerbated their hyper-exponential shape in finer sediments. Furthermore, the amounts of MPs present in the effluent increased to over 85 % as MPs size decreased to 10-20 µm in both gravel and coarse sand columns, while all larger MPs (125-150 µm) were retained in the coarse sand column. The modeling results suggested that the blocking mechanism becomes more important with increasing particle sizes. In particular, the attachment-detachment without blocking was the most suited parameterization to interpret the movement of small MPs, while a depth-dependent blocking approach was necessary to adequately describe the fate of larger particles.

Multiplexed Near-Field Optical Trapping Exploiting Anapole States.

Optical tweezers have had a major impact on bioscience research by enabling the study of biological particles with high accuracy. The focus so far has been on trapping individual particles, ranging from the cellular to the molecular level. However, biology is intrinsically heterogeneous; therefore, access to variations within the same population and species is necessary for the rigorous understanding of a biological system. Optical tweezers have demonstrated the ability of trapping multiple targets in parallel; however, the multiplexing capability becomes a challenge when moving toward the nanoscale. Here, we experimentally demonstrate a resonant metasurface that is capable of trapping a high number of nanoparticles in parallel, thereby opening up the field to large-scale multiplexed optical trapping. The unit cell of the metasurface supports an anapole state that generates a strong field enhancement for low-power near-field trapping; importantly, the anapole state is also more angle-tolerant than comparable resonant modes, which allows its excitation with a focused light beam, necessary for generating the required power density and optical forces. We use the anapole state to demonstrate the trapping of 100's of 100 nm polystyrene beads over a 10 min period, as well as the multiplexed trapping of lipid vesicles with a moderate intensity of <250 µW/µm2. This demonstration will enable studies relating to the heterogeneity of biological systems, such as viruses, extracellular vesicles, and other bioparticles at the nanoscale.

Estimating the impact of vadose zone heterogeneity on agricultural managed aquifer recharge: A combined experimental and modeling study.

Agricultural managed aquifer recharge (Ag-MAR) is a promising approach to replenish groundwater resources using flood water and cropland as spreading grounds. However, site selection, particularly the layering of sediment deposits in the subsurface, can greatly influence Ag-MAR efficacy as it controls water flow and solute transport in the vadose zone. In this study, we use the HYDRUS-1D software to simulate water flow and solute transport from the land surface to the groundwater table in three vadose zone profiles (LS, MS, HS) characterized by differing fractions of sand (44 %, 47 %, and 64 %). For each profile, the single- and dual-porosity models (i.e., considering or not nonequilibrium water flow and solute transport) were calibrated using observed surface ponding, soil water content, and KBr breakthrough data. Water flow and bromide transport in the profile with the lowest sand fraction (LS) were best captured using the model that considered both preferential flow and nonequilibrium bromide transport. Water flow and bromide transport in the profile with the highest sand fraction (HS) was best simulated with the model that considered preferential flow and equilibrium bromide transport. Uniform water flow and nonequilibrium bromide transport provided the best fit for the third profile (MS). The degree of preferential flow was highest in the profile with the largest sand fraction (HS), which also showed the largest flow velocities compared to the profiles with lower sand amounts (LS and MS). Preferential flow did not significantly impact the overall water balance (within 3 %), but caused a significant decrease in vadose zone travel times (bromide) by up to 38 %, relative to a single-porosity model fit. Recharge efficiency varied between 88 % and 90 %, while the average travel times from the soil surface to groundwater varied up to 119 % (from 3.6 to 7.9 days) between the three sites. This study demonstrates that similar recharge efficiency can be achieved at sites with differing soil texture profiles, but subsurface heterogeneity can substantially affect contaminant transport processes and their travel times.

Highly Sensitive Refractive Index Sensor Based on Polymer Bragg Grating: A Case Study on Extracellular Vesicles Detection.

The measurement of small changes in the refractive index (RI) leads to a comprehensive analysis of different biochemical substances, paving the way to non-invasive and cost-effective medical diagnosis. In recent times, the liquid biopsy for cancer detection via extracellular vesicles (EV) in the bodily fluid is becoming very popular thanks to less invasiveness and stability. In this context, here we propose a highly sensitive RI sensor based on a compact high-index-coated polymer waveguide Bragg grating with a metal under cladding. Owing to the combined effect of a metal under cladding and a high-index coating, a significant enhancement in the RI sensitivity as well as the dynamic range has been observed. The proposed sensor has been analyzed by combining finite element method (FEM) and coupled-mode theory (CMT) approaches, demonstrating a sensitivity of 408-861 nm/RIU over a broad dynamic range of 1.32-1.44, and a strong evanescent field within a 150 nm proximity to the waveguide surface compliant with EV size. The aforementioned performance makes the proposed device suitable for performing real-time and on-chip diagnoses of cancer in the early stage.

A novel multiscale biophysical model to predict the fate of ionizable compounds in the soil-plant continuum.

Soil pollution from emerging contaminants poses a significant threat to water resources management and food production. The development of numerical models to describe the reactive transport of chemicals in both soil and plant is of paramount importance to elaborate mitigation strategies. To this aim, in the present study, a multiscale biophysical model is developed to predict the fate of ionizable compound in the soil-plant continuum. The modeling framework connects a multi-organelles model to describe processes at the cell level with a semi-mechanistic soil-plant model, which includes the widely used Richards-based solver, HYDRUS. A Bayesian probabilistic framework is used to calibrate and assess the capability of the model in reproducing the observations from an experiment on the translocation of five pharmaceuticals in green pea plants. Results show satisfactory fitting performance and limited predictive uncertainty. The subsequent validation with the cell model indicates that the estimated soil-plant parameters preserve a physically realistic meaning, and their calibrated values are comparable with the existing literature values, thus confirming the overall reliability of the analysis. Model results further suggest that pH conditions in both soil and xylem play a crucial role in the uptake and translocation of ionizable compounds.

Novel Micro-Nano Optoelectronic Biosensor for Label-Free Real-Time Biofilm Monitoring.

According to the World Health Organization forecasts, AntiMicrobial Resistance (AMR) is expected to become one of the leading causes of death worldwide in the following decades. The rising danger of AMR is caused by the overuse of antibiotics, which are becoming ineffective against many pathogens, particularly in the presence of bacterial biofilms. In this context, non-destructive label-free techniques for the real-time study of the biofilm generation and maturation, together with the analysis of the efficiency of antibiotics, are in high demand. Here, we propose the design of a novel optoelectronic device based on a dual array of interdigitated micro- and nanoelectrodes in parallel, aiming at monitoring the bacterial biofilm evolution by using optical and electrical measurements. The optical response given by the nanostructure, based on the Guided Mode Resonance effect with a Q-factor of about 400 and normalized resonance amplitude of about 0.8, allows high spatial resolution for the analysis of the interaction between planktonic bacteria distributed in small colonies and their role in the biofilm generation, calculating a resonance wavelength shift variation of 0.9 nm in the presence of bacteria on the surface, while the electrical response with both micro- and nanoelectrodes is necessary for the study of the metabolic state of the bacteria to reveal the efficacy of antibiotics for the destruction of the biofilm, measuring a current change of 330 nA when a 15 µm thick biofilm is destroyed with respect to the absence of biofilm.

Characteristics of COVID-19 Pneumonia Survivors With Resting Normoxemia and Exercise-Induced Desaturation.

BACKGROUND: Survivors of coronavirus disease 2019 (COVID-19) associated pneumonia may show exercise-induced desaturation. We wondered whether these individuals show physiologic and symptom characteristics similar to individuals with chronic respiratory diseases with exercise-induced desaturation, namely COPD or interstitial lung diseases (ILD). We evaluated lung function, exercise capacity, and symptoms in these individuals compared with individuals with COPD or ILD and exercise-induced desaturation. METHODS: Survivors of COVID-19 associated pneumonia (study individuals), normoxemic at rest with exercise-induced desaturation, underwent assessment of dyspnea, dynamic lung volumes, carbon monoxide diffusion capacity, and the 6-min walk test. Data of individuals with COPD or with ILD and exercise-induced desaturation were also retrospectively analyzed. RESULTS: FVC was lower in individuals with COVID-19 or ILD than in those with COPD. Individuals who had COVID-19 walked < 70% of predicted and, as a whole, had a 6-min walk test performance similar to individuals with ILD but walked significantly less, showed more severe leg fatigue and dyspnea during exercise, and more exercise-induced desaturation than individuals with COPD. CONCLUSIONS: Survivors of COVID-19 associated pneumonia, who were normoxemic at rest with exercise-induced desaturation, had alterations in lung function, exercise capacity, and symptoms similar to individuals with ILD but more severe than individuals with COPD and exercise-induced desaturation.

Pulmonary rehabilitation in patients with interstitial lung diseases: Correlates of success.

BACKGROUND AND AIM: Benefits of pulmonary rehabilitation in Interstitial Lung Diseases (ILD) have been reported. The aim of this large multicenter study was to identify the success predictors of pulmonary rehabilitation in a real-life setting. METHODS: Data of 240 in-patients (110 idiopathic pulmonary fibrosis (IPF), 106 ILD other than IPF and 24 undetermined ILD) undergoing pulmonary rehabilitation in a 10-year period were retrospectively evaluated. Six minute walking distance (6MWT), body weight-walking distance product tests, dyspnoea and arterial blood gases were assessed at admission and discharge. Differences in post rehabilitation changes in outcome measures as function of baseline characteristics were evaluated. RESULTS: After rehabilitation, patients showed improvements in all outcome measures (p < 0.05), regardless of the underlying diagnosis or disease severity. Patients needing oxygen therapy at rest showed reduced benefits. Baseline 6MWD inversely correlated with its changes at discharge. Non-significant greater benefits after rehabilitation were found in IPF patients under antifibrotic therapy. In a subset of 50 patients assessed on average 10.3 ± 3.5 months after discharge, the benefits in 6MWD were not maintained (312.9 ± 139.4, 369.7 ± 122.5 and 310.8 ± 139.6 m at admission, discharge and follow up respectively: p < 0.0001). CONCLUSION: Pulmonary rehabilitation may improve dyspnoea, exercise capacity and fatigue in patients with ILD of different aethiologies and level of severity. The long-term effects need to be established.

The cAMP-HMGA1-RBP4 system: a novel biochemical pathway for modulating glucose homeostasis.

BACKGROUND: We previously showed that mice lacking the high mobility group A1 gene (Hmga1-knockout mice) developed a type 2-like diabetic phenotype, in which cell-surface insulin receptors were dramatically reduced (below 10% of those in the controls) in the major targets of insulin action, and glucose intolerance was associated with increased peripheral insulin sensitivity. This particular phenotype supports the existence of compensatory mechanisms of insulin resistance that promote glucose uptake and disposal in peripheral tissues by either insulin-dependent or insulin-independent mechanisms. We explored the role of these mechanisms in the regulation of glucose homeostasis by studying the Hmga1-knockout mouse model. Also, the hypothesis that increased insulin sensitivity in Hmga1-deficient mice could be related to the deficit of an insulin resistance factor is discussed. RESULTS: We first show that HMGA1 is needed for basal and cAMP-induced retinol-binding protein 4 (RBP4) gene and protein expression in living cells of both human and mouse origin. Then, by employing the Hmga1-knockout mouse model, we provide evidence for the identification of a novel biochemical pathway involving HMGA1 and the RBP4, whose activation by the cAMP-signaling pathway may play an essential role for maintaining glucose metabolism homeostasis in vivo, in certain adverse metabolic conditions in which insulin action is precluded. In comparative studies of normal and mutant mice, glucagon administration caused a considerable upregulation of HMGA1 and RBP4 expression both at the mRNA and protein level in wild-type animals. Conversely, in Hmga1-knockout mice, basal and glucagon-mediated expression of RBP4 was severely attenuated and correlated inversely with increased Glut4 mRNA and protein abundance in skeletal muscle and fat, in which the activation state of the protein kinase Akt, an important downstream mediator of the metabolic effects of insulin on Glut4 translocation and carbohydrate metabolism, was simultaneously increased. CONCLUSION: These results indicate that HMGA1 is an important modulator of RBP4 gene expression in vivo. Further, they provide evidence for the identification of a novel biochemical pathway involving the cAMP-HMGA1-RBP4 system, whose activation may play a role in glucose homeostasis in both rodents and humans. Elucidating these mechanisms has importance for both fundamental biology and therapeutic implications.

Disentangling model complexity in green roof hydrological analysis: A Bayesian perspective.

Green Roofs (GRs) have proven to be a sustainable solution to stormwater management in urban areas. To boost their adoption at the large scale, there is a need to develop numerical models, which are accurate, computationally cheap, and as complex as needed to reproduce the hydrological behavior of GRs. Alternative conceptual and mechanistic approaches have been proposed and tested, however the most appropriate level of model complexity for GRs' analysis is still unknown. To cover this scientific gap, we provide a Bayesian comprehensive perspective of GR hydrological modeling, which includes a statistically rigorous Bayesian comparison of one conceptual and multiple Richards-based mechanistic GR models, and a probabilistic assessment of the information content of different observations. The analysis of the marginal likelihoods reveals that the conceptual and the unimodal van Genuchten - Mualem models are the most appropriate parameterizations, and that further layers of model complexity are not fully supported by the measurements. In addition to that, the estimated Kullback-Leibler divergences suggest that the measured volumetric water content outperforms the measured subsurface outflow and tracer concentrations in terms of informativeness, leading to the lowest model predictive uncertainty for the simulation of water fluxes. The findings of this study represent a first step to clarify the role of model complexity in GRs' analysis, and open new perspective on GRs' model-based experimental design.

Monitoring of individual bacteria using electro-photonic traps.

Antimicrobial resistance (AMR) describes the ability of bacteria to become immune to antimicrobial treatments. Current testing for AMR is based on culturing methods that are very slow because they assess the average response of billions of bacteria. In principle, if tests were available that could assess the response of individual bacteria, they could be much faster. Here, we propose an electro-photonic approach for the analysis and the monitoring of susceptibility at the single-bacterium level. Our method employs optical tweezers based on photonic crystal cavities for the trapping of individual bacteria. While the bacteria are trapped, antibiotics can be added to the medium and the corresponding changes in the optical properties and motility of the bacteria be monitored via changes of the resonance wavelength and transmission. Furthermore, the proposed assay is able to monitor the impedance of the medium surrounding the bacterium, which allows us to record changes in metabolic rate in response to the antibiotic challenge. For example, our simulations predict a variation in measurable electrical current of up to 40% between dead and live bacteria. The proposed platform is the first, to our knowledge, that allows the parallel study of both the optical and the electrical response of individual bacteria to antibiotic challenge. Our platform opens up new lines of enquiry for monitoring the response of bacteria and it could lead the way towards the dissemination of a new generation of antibiogram study, which is relevant for the development of a point-of-care AMR diagnostics.

GPER Mediates a Feedforward FGF2/FGFR1 Paracrine Activation Coupling CAFs to Cancer Cells toward Breast Tumor Progression.

The FGF2/FGFR1 paracrine loop is involved in the cross-talk between breast cancer cells and components of the tumor stroma as cancer-associated fibroblasts (CAFs). By quantitative PCR (qPCR), western blot, immunofluorescence analysis, ELISA and ChIP assays, we demonstrated that 17ß-estradiol (E2) and the G protein estrogen receptor (GPER) agonist G-1 induce the up-regulation and secretion of FGF2 via GPER together with the EGFR/ERK/c-fos/AP-1 signaling cascade in (ER)-negative primary CAFs. Evaluating the genetic alterations from METABRIC and TCGA datasets, we then assessed that FGFR1 is the most frequently amplified FGFRs family member and its amplification/expression associates with shorter survival rates in breast cancer patients. Therefore, in order to assess the functional FGF2/FGFR1 interplay between CAFs and breast cancer cells, we generated the FGFR1-knockout MDA-MB-231 cells using CRISPR/Cas9 genome editing strategy. Using conditioned medium from estrogen-stimulated CAFs, we established that the activation of FGF2/FGFR1 paracrine signaling triggers the expression of the connective tissue growth factor (CTGF), leading to the migration and invasion of MDA-MB-231 cells. Our findings shed new light on the role elicited by estrogens through GPER in the activation of the FGF2/FGFR1 signaling. Moreover, our findings may identify further biological targets that could be considered in innovative combination strategies halting breast cancer progression.

Comparison between exhaled and bronchoalveolar lavage levels of hydrogen peroxide in patients with diffuse interstitial lung diseases.

The aim of the present study was to compare hydrogen peroxide (H2O2) levels resulting from oxidative stress in exhaled breath condensate (EBC) and bronchoalveolar lavage (BAL) supernatants of subjects with different diffuse interstitial lung diseases (DILDs). Twenty-one patients who underwent BAL procedure for various DILDs were studied. EBC, which was collected the day before bronchoscopy, was obtained by cooling exhaled breath; BAL was processed for differential cellular count and supernatant was stored. H2O2 in both fluids was measured using a commercial fluorimetric kit with a limit of detection of 0.01 microM. No difference in H2O2 levels in EBC and in BAL was observed [median (range), 0.07 microM (0.01-0.6) and 0.08 microM (0.01-0.8), respectively]. No correlation was observed between BAL and EBC levels. H2O2 in BAL did not correlate with differential cellular count, whereas H2O2 EBC did correlate positively with the percentage of epithelial cells (r = 0.5, p = 0.007) and negatively with the number of macrophages (r = -0.4, p = 0.03). No correlation was observed between H2O2 levels either in EBC or BAL and lung function data. The data show that in DILDs H2O2 can be detected in both EBC and supernatants of BAL and that their relative concentrations are similar but not correlated with each other. The positive correlation between H2O2 levels in EBC and percentage of epithelial cells leads to the speculation that airway epithelia may play a relevant contribution in H2O2 production in the airway lumen.