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Getting paid has never been so complicated for primary care doctors. An uncertain policy landscape, ever-changing regulatory requirements, and evolving practice and reimbursement models mean that physicians must constantly stay [...]

Abstract Mechanisms that control progression from simple steatosis to steato-hepatitis and fibrosis in patients with non-alcoholic fatty liver disease (NAFLD) are unknown. SPARC, a secreted matricellular protein, is over-expressed in the liver under chronic injury. Contribution of SPARC accumulation to disease severity is largely unknown in NAFLD. We assessed the hypothesis that SPARC is increased in livers with more necrosis and inflammation and could be associated with more fibrosis. qrt-PCR, immunohistochemistry, and ELISA were employed to localize and quantify changes in SPARC in 62 morbidly obese patients with NAFLD/NASH and in a mouse model of diet-induced-NASH. Results were correlated with the severity of NAFLD/NASH. In obese patients 2 subgroups were identified with either high SPARC expression (n = 16) or low SPARC expression (n = 46) in the liver, with a cutoff of 1.2 fold expression. High expression of SPARC paralleled hepatocellular damage and increased mRNA expression of pro-fibrogenic factors in the liver. In line with these findings, in the NASH animal model SPARC knockout mice were protected from inflammatory injury, and showed less inflammation and fibrosis. Hepatic SPARC expression is associated with liver injury and fibrogenic processes in NAFLD. SPARC has potential as preventive or therapeutic target in NAFLD patients.

Abstract Cyclic nucleotide-gated (CNG) channels mediate transduction in several sensory neurons. These channels use the free energy of CNs’ binding to open the pore, a process referred to as gating. CNG channels belong to the superfamily of voltage-gated channels, where the motion of the α-helix S6 controls gating in most of its members. To date, only the open, cGMP-bound, structure of a CNG channel has been determined at atomic resolution, which is inadequate to determine the molecular events underlying gating. By using electrophysiology, site-directed mutagenesis, chemical modification, and Single Molecule Force Spectroscopy, we demonstrate that opening of CNGA1 channels is initiated by the formation of salt bridges between residues in the C-linker and S5 helix. These events trigger conformational changes of the α-helix S5, transmitted to the P-helix and leading to channel opening. Therefore, the superfamily of voltage-gated channels shares a similar molecular architecture but has evolved divergent gating mechanisms.

Alessandro Zanasi,1 Luigi Morcaldi,2 Salvatore Cazzato,3 Massimiliano Mazzolini,4 Marzia Lecchi,5 Antonio Maria Morselli-Labate,6 Marianna Mastroroberto,6 Roberto W Dal Negro7 1Italian Association for Cough Study, Bologna, 2National Observatory of Health in Childhood and Adolescence (PAIDOSS: Osservatorio Nazionale sulla Salute dell’Infanzia e dell’Adolescenza), Rome, 3Department of Pediatrics, Salesi Children’s Hospital, Ancona, 4Department of Specialist, Diagnostic and Experimental Medicine, Respiratory and Critical Care Unit, S.Orsola-Malpighi Hospital, University of Bologna, Bologna, 5Department of Biotechnology and Biosciences, University of Milan Bicocca, Milan, 6Department of Medical and Surgical Sciences, Alma Mater Studiorum, University of Bologna, Bologna, 7National Centre for Respiratory Pharmacoeconomics and Pharmacoepidemiology, Verona, Italy Context: Children’s cough is a daily concern for most pediatricians. The management of both acute and chronic cough requires a systematic and comprehensive approach. Despite the approved protocols for management, the pediatric assessment of cough and the corresponding prescribing attitude frequently do not fit these protocols, which can be affected by parental suggestions – sometimes substantially. Objective: The objective of this study was to investigate both the perception and the behavior of a representative sample of Italian pediatricians toward cough in real life. Methods: A specific questionnaire consisting of 18 questions was prepared. The questionnaire was completed by 300 pediatricians (all members of PAIDOSS: Italian National Observatory on Health of Childhood and Adolescence) who represented ~300,000 children. Results: A vast majority of children have cough throughout the year (99.3% of respondents have cough during autumn/winter and 64.7% in spring/summer). Allergic disease is the most frequent suspected cause of chronic cough in children (53%), and this is supported by the high demand for consultations: 73% seek the opinion of allergologists, 62% of otorhinolaryngologists and only 33% of pulmonologists. The majority of pediatricians (92%) reported that they prescribe therapy in acute cough regardless of cough guidelines. Moreover, the survey pointed out the abuse of aerosol therapy (26% in acute cough and 38% in chronic cough) and of antibiotics prescription (22% in acute cough and 42% in chronic cough). Conclusion: Our survey suggests that some Italian pediatricians’ therapeutic attitudes should be substantially improved in order to achieve better management of cough in children and to minimize the burden of cough. Keywords: acute cough, chronic cough, cough in children, pediatricians’ prescriptions, cough impact

We experimentally demonstrate how In-mediated metal-exchange catalysis (MEXCAT) allows us to widen the deposition window for β-Ga2O3 homoepitaxy to conditions otherwise prohibitive for its growth via molecular beam epitaxy (e.g., substrate temperatures ≥800 °C) on the major substrate orientations, i.e., (010), (001), (2¯01), and (100) 6°-offcut. The obtained crystalline qualities, surface roughnesses, growth rates, and In-incorporation profiles are shown and compared with different experimental techniques. The growth rates, Γ, for fixed growth conditions are monotonously increasing with the surface free energy of the different orientations with the following order: Γ(010) > Γ(001) > Γ(2¯01) > Γ(100). Ga2O3 surfaces with higher surface free energy provide stronger bonds to the surface ad-atoms or ad-molecules, resulting in decreasing desorption, i.e., a higher incorporation/growth rate. The structural quality in the case of (2¯01), however, is compromised by twin domains due to the crystallography of this orientation. Notably, our study highlights β-Ga2O3 layers with high structural quality grown by MEXCAT-MBE not only in the most investigated (010) orientation but also in the (100) and (001) ones. In particular, MEXCAT on the (001) orientation results in both growth rate and structural quality comparable to the ones achievable with (010), and the limited incorporation of In associated with the MEXCAT deposition process does not change the insulating characteristics of unintentionally doped layers. The (001) surface is therefore suggested as a valuable alternative orientation for devices.

Previously we described direct cellular interactions between microglia and AKT1+ brain tumour cells in zebrafish (Chia et al., 2018). However, it was unclear how these interactions were initiated: it was also not clear if they had an impact on the growth of tumour cells. Here, we show that neoplastic cells hijack mechanisms that are usually employed to direct microglial processes towards highly active neurons and injuries in the brain. We show that AKT1+ cells possess dynamically regulated high intracellular Ca2+ levels. Using a combination of live imaging, genetic and pharmacological tools, we show that these Ca2+ transients stimulate ATP-mediated interactions with microglia. Interfering with Ca2+ levels, inhibiting ATP release and CRISPR-mediated mutation of the p2ry12 locus abolishes these interactions. Finally, we show that reducing the number of microglial interactions significantly impairs the proliferation of neoplastic AKT1 cells. In conclusion, neoplastic cells repurpose the endogenous neuron to microglia signalling mechanism via P2ry12 activation to promote their own proliferation.