Gene identification for risk of relapse in stage I lung adenocarcinoma patients. A combined methodology of gene expression profiling and computational gene network analysis

Risk assessment and treatment choice remains a challenge in early non-smallcell lung cancer (NSCLC). The aim of this study was to identify novel genes involved in the risk of early relapse (ER) compared to no relapse (NR) in resected lung adenocarcinoma (AD) patients using a combination of high throughput technology and computational analysis. We identified 18 patients (n.13 NR and n.5 ER) with stage I AD. Frozen samples of patients in ER, NR and corresponding normal lung (NL) were subjected to Microarray technology and quantitative-PCR (Q-PCR). A gene network computational analysis was performed to select predictive genes. An independent set of 79 ADs stage I samples was used to validate selected genes by Q-PCR. From microarray analysis we selected 50 genes, using the fold change ratio of ER versus NR. They were validated both in pool and individually in patient samples (ER and NR) by Q-PCR. Fourteen increased and 25 decreased genes showed a concordance between two methods. They were used to perform a computational gene network analysis that identified 4 increased (HOXA10, CLCA2, AKR1B10, FABP3) and 6 decreased (SCGB1A1, PGC, TFF1, PSCA, SPRR1B and PRSS1) genes. Moreover, in an independent dataset of ADs samples, we showed that both high FABP3 expression and low SCGB1A1 expression was associated with a worse disease-free survival (DFS). Our results indicate that it is possible to define, through gene expression and computational analysis, a characteristic gene profiling of patients with an increased risk of relapse that may become a tool for patient selection for adjuvant therapy

Preprocedural Level of Soluble CD40L Is Predictive of Enhanced Inflammatory Response and Restenosis After Coronary Angioplasty

Background— Inflammation plays a pathogenic role in the development of restenosis after percutaneous transluminal coronary angioplasty (PTCA). CD40–CD40L interaction is involved in the pathogenesis of atherosclerosis; however, its role in the pathophysiology of restenosis is still unclear. We tested the hypothesis that soluble CD40L (sCD40L) may be involved in the process of restenosis and that it exerts its effect by triggering a complex group of inflammatory reactions on endothelial and mononuclear cells. Methods and Results— We studied 70 patients who underwent PTCA and who had repeated angiograms at 6-month follow-up. Plasma sCD40L was measured before and 1, 5, 15, and 180 days after PTCA, whereas plasma soluble intercellular adhesion molecule-1, soluble vascular cell adhesion molecule-1, E-selectin, and monocyte chemoattractant protein (MCP)-1 were measured before and 24 hours after PTCA. Furthermore, the release of adhesion molecules and MCP-1 and the ability to repair an injury in endothelial cells, as well as the generation of O 2 − in monocytes, were analyzed in vitro after stimulation with serum from patients or healthy control subjects. Restenosis occurred in 18 patients (26%). Restenotic patients had preprocedural sCD40L significantly higher than patients with favorable outcomes (2.13±0.3 versus 0.87±0.12 ng/mL, P <0.0001). Elevated sCD40L at baseline was significantly correlated with adhesion molecules and MCP-1 generation after PTCA and with lumen loss at 6-month follow-up. Furthermore, high sCD40L was directly associated in vitro with adhesion molecules and MCP-1 generation and impaired migration in endothelial cells and with enhanced O 2 − generation in monocytes. Conclusions— We conclude that increased sCD40L is associated with late restenosis after PTCA. This may provide an important biochemical link between restenosis and aspirin-insensitive platelet activation. These results provide a rationale for studies with new antiplatelet treatments in patients who underwent PTCA

Phenolic extraction of Moringa oleifera leaves in DES: characterization of the extracts and their application in methylcellulose films for food packaging

In this work, a qualitative study of the phenolic content of Moringa oleifera leaves (MO), extracted with deep eutectic solvents (DES) based on choline chloride (ChCl) with lactic acid (LA) or glycerol (GLY), was performed by high-resolution mass spectrometry (HPLC-DAD-ESI-MSn). The two solvents (DES-LA and DES-GLY) extract similar classes of phenolics, and ten compounds were identified. The antioxidant profile was also studied (TPC, TFC, DPPH, FRAP, ORAC, and ABTS). Both solvents show an efficient extraction of phenolic compounds and high antioxidant capacity was verified for the extracts. However, the DES-Gly have a higher capacity for polyphenolic extraction (TPC led to 38.409 ± 0.095 mg GAE.g−1 and 2.259 ± 0.023 mg QE.g−1 for TFC). Films based on methylcellulose (MC) containing different amounts of DES or MO extracts, acting as plasticizers, were developed and characterized regarding their mechanical, optical, water vapor permeability, and microstructural properties. All films are uniform, clear, and transparent with smooth, homogeneous surfaces. It was found that the presence of more than 10% of MO extract and/or DES provided more flexible films (Eb for MC 2%_DES 20% achieved 4.330 ± 0.27 %, and 8.15 ± 0.39 % for MC 2%_MO 20%) with less mechanical and barrier resistance. The ultimate objective of this study was to provide information that could assist in the development of antimicrobial active methylcellulose films for sliced wheat bread packaging.Projects UIDB/50006/2020 and UIDP/50006/2020, funded by FCT/MCTES through national funds

Intra-and extra-hospitalization monitoring of vital signs. Two sides of the same coin. Perspectives from Lims and Greenline study operators

Background: In recent years, due to the epidemiological transition, the burden of very complex patients in hospital wards has increased. Telemedicine usage appears to be a potential high-impact factor in helping with patient management, allowing hospital personnel to assess conditions in out-of-hospital scenarios. Methods: To investigate the management of chronic patients during both hospitalization for disease and discharge, randomized studies (LIMS and Greenline-HT) are ongoing in the Internal Medicine Unit at ASL Roma 6 Castelli Hospital. The study endpoints are clinical outcomes (from a patient’s perspective). In this perspective paper, the main findings of these studies, from the operators’ point of view, are reported. Operator opinions were collected from structured and unstructured surveys conducted among the staff involved, and their main themes are reported in a narrative manner. Results: Telemonitoring appears to be linked to a reduction in side-events and side-effects, which represent some of most commons risk factors for re-hospitalization and for delayed discharge during hospitalization. The main perceived advantages are increased patient safety and the quick response in case of emergency. The main disadvantages are believed to be related to low patient compliance and an infrastructural lack of optimization. Conclusions: The evidence of wireless monitoring studies, combined with the analysis of activity data, suggests the need for a model of patient management that envisages an increase in the territory of structures capable of offering patients subacute care (the possibility of antibiotic treatments, blood transfusions, infusion support, and pain therapy) for the timely management of chronic patients in the terminal phase, for which treatment in acute wards must be guaranteed only for a limited time for the management of the acute phase of their diseases

Time to Switch to Second-line Antiretroviral Therapy in Children With Human Immunodeficiency Virus in Europe and Thailand.

Background: Data on durability of first-line antiretroviral therapy (ART) in children with human immunodeficiency virus (HIV) are limited. We assessed time to switch to second-line therapy in 16 European countries and Thailand. Methods: Children aged <18 years initiating combination ART (≥2 nucleoside reverse transcriptase inhibitors [NRTIs] plus nonnucleoside reverse transcriptase inhibitor [NNRTI] or boosted protease inhibitor [PI]) were included. Switch to second-line was defined as (i) change across drug class (PI to NNRTI or vice versa) or within PI class plus change of ≥1 NRTI; (ii) change from single to dual PI; or (iii) addition of a new drug class. Cumulative incidence of switch was calculated with death and loss to follow-up as competing risks. Results: Of 3668 children included, median age at ART initiation was 6.1 (interquartile range (IQR), 1.7-10.5) years. Initial regimens were 32% PI based, 34% nevirapine (NVP) based, and 33% efavirenz based. Median duration of follow-up was 5.4 (IQR, 2.9-8.3) years. Cumulative incidence of switch at 5 years was 21% (95% confidence interval, 20%-23%), with significant regional variations. Median time to switch was 30 (IQR, 16-58) months; two-thirds of switches were related to treatment failure. In multivariable analysis, older age, severe immunosuppression and higher viral load (VL) at ART start, and NVP-based initial regimens were associated with increased risk of switch. Conclusions: One in 5 children switched to a second-line regimen by 5 years of ART, with two-thirds failure related. Advanced HIV, older age, and NVP-based regimens were associated with increased risk of switch

Coupling cAMP Signaling to Transcription in the Liver: Pivotal Role of CREB and CREM

International audienceTranscriptional factors binding to cAMP-responsive elements (CREs) in the promoters of various genes belong to the basic domain-leucine zipper superfamily and are composed of three genes in mammals, CREB, CREM, and ATF-1. A large number of CREB, CREM, and ATF-1 proteins are generated by posttranscriptional events, mostly alternative splicing, and regulate gene expression by acting as activators or repressors. Activation is classically brought about by signaling-dependent phosphorylation of a key acceptor site (Ser133 in CREB) by a number of possible kinases, including PKA, CamKIV, and Rsk-2. Phosphorylation is the prerequisite for the interaction of CBP (CREB-binding protein), a co-activator that has also histone acetyltransferase activity. Repression may involve dynamic dephosphorylation of the activators and thus decreased association with CBP. Another pathway of transcriptional repression on CRE sites implicates the inducible repressor ICER (inducible cAMP early repressor), a product of the CREM gene. Being an inducible repressor, ICER is involved in autoregulatory feedback loops of transcription that govern the down-regulation of early response genes, such as the proto-oncogene c-fos. The liver represents a remarkable physiological setting where cAMP-responsive signaling plays a major role. Indeed, a finely tuned program of gene expression is triggered by partial hepatectomy, so that through specific checkpoints a coordinated regeneration of the tissue is obtained. Temporal kinetics of transcriptional activation after hepatectomy reveals a pattern of early induction for several genes, some of them controlled by the CREB/CREM transcription factors. An important role of CREM in liver physiology was suggested by the robust induction of ICER after partial hepatectomy. The delay in tissue regeneration in CREM-deficient mice confirmed the important function of this factor in regulating hepatocyte proliferation. As gene induction is accompanied by critical changes in chromatin organization, the deciphering of the specific modification codes that histones display during liver regeneration and physiology will provide exciting new insights into the dynamics of chromatin architecture

Transcription factor CREM coordinates the timing of hepatocyte proliferation in the regenerating liver

The liver regenerates upon partial hepatectomy (PH) as terminally differentiated hepatocytes undergo a tremendous proliferative process. CREM gene expression is powerfully induced during liver regeneration. We show that cell proliferation is significantly reduced upon PH in CREM(−/−) mice. There is a reduction in DNA synthesis, in the number of mitosis and of phosphorylated histone H3-positive cells. The post-PH proliferation peak is delayed by 10 hr, indicating an altered hepatocyte cell cycle. Expression of cyclins A, B, D1, E, and cdc2, of c-fos and tyrosine aminotransferase is deregulated. CREM mutation results in delayed S-phase entry, impairing the synchronization of proliferation

An Interface Model Including Cracks and Roughness Applied to Masonry

International audienceIn this paper an interface model accounting for roughness and micro-cracks is presented and applied to masonry-like structures. The model is consistently derived by coupling a homogenization approach and arguments of asymptotic analyses. A numerical procedure is introduced and numerical results, based on a finite element formulation, are successfully compared with experimental data , obtained on masonry samples undergoing to shear tests. Finally, a parametric numerical analysis is proposed, highlighting the influence of the roughness features on the interface response

Reconstruction of the dynamic exposure to PCBs for Italian population using physiologically based pharmacokinetic (PBPK) model and a complex approach combining human biomonitoring studies and environmental data

Polychlorinated biphenyls (PCBs) are persistent contaminants suspected to cause adverse health effects in humans. Due to their extensive use in the past and persistence in the environment significant amounts are still detectable in environmental media. The major route of human exposure is contaminated food. Since food contamination by PCBs has not been monitored frequently in the past and was rarely congener-specific, human exposure cannot be directly assessed. An alternative to this lack of data is to use exposure models developed for environmental media and to transpose the environmental model outcomes into time-variant dietary intakes. According to the hypotheses assumed, the shape and the magnitude of the modeled intakes are affected by uncertainty leading to several possible exposure scenarios. Here we propose to improve this methodology by integrating actual internal levels in humans (from biomonitoring studies) in order to reduce the uncertainty in the exposure scenario. We propose to model the exposure and the toxicokinetics of PCBs in humans and to analyze breast milk levels of Italian primiparous women in this modeling framework in order to determine the most likely scenario of exposure (i.e. the shape and magnitude). Our approach was applied to most prevalent PCB congeners detected in human milk and fluids: PCB-138 (2,2',3,4,4',5'-hexachlorobiphenyl), PCB-153 (2,2',4,4',5,5'-hexachlorobiphenyl) and PCB-180 (2,2',3,4,4',5,5' -heptachlorobiphenyl). Breast milk concentrations were measured in Italian women for PCB-138, PCB-153 and PCB-180. Forty one healthy primiparous women, not-occupationally exposed to PCBs, were included in biomonitoring study: 15 women from Giugliano in Campania, 18 from Piacenza and 8 from Milan. For each congener, three exposure scenarios were derived and a PBPK model was used to relate the lifetime exposure to the breast milk levels. For the three PCBs, we determined the most probable scenario of exposure. A Bayesian analysis was performed to estimate the magnitude of each exposure scenario for each woman using the breast milk concentrations. The intake estimates are in good agreement with previous exposure assessments based solely on food contamination demonstrating the relevance of our approach to reconstruct accurately the exposure and to fill in data gaps on exposure. The link between the exposure scenario and biomonitoring data was provided by the PBPK model in order to support the adequacy of the models according to the data (e.g., for PCB-180) or to identify possible improvements for the exposure and PBPK models (e.g., for PCB-138 and PCB-153)