Microenvironment inflammatory infiltrate drives growth speed and outcome of hepatocellular carcinoma: a prospective clinical study

In HCC, tumor microenvironment, heavily influenced by the underlying chronic liver disease, etiology and stage of the tissue damage, affects tumor progression and determines the high heterogeneity of the tumor. Aim of this study was to identify the circulating and tissue components of the microenvironment immune-mediated response affecting the aggressiveness and the ensuing clinical outcome. We analyzed the baseline paired HCC and the surrounding tissue biopsies from a prospective cohort of 132 patients at the first diagnosis of HCC for immunolocalization of PD-1/PD-L1, FoxP3, E-cadherin, CLEC2 and for a panel of 82 microRNA associated with regulation of angiogenesis, cell proliferation, cell signaling, immune control and autophagy. Original microarray data were also explored. Serum samples were analyzed for a panel of 19 cytokines. Data were associated with biochemical data, histopathology and survival. Patients with a more aggressive disease and shorter survival, who we named fast-growing accordingly to the tumor doubling time, at presentation had significantly higher AFP levels, TGF-β1 and Cyphra 21-1 levels. Transcriptomic analysis evidenced a significant downregulation of CLEC2 and upregulation of several metalloproteinases. A marked local upregulation of both PD-1 and PD-L1, a concomitant FoxP3-positive lymphocytic infiltrate, a loss of E-cadherin, gain of epithelial-mesenchymal transition (EMT) phenotype and extreme poor differentiation at histology were also present. Upregulated microRNA in fast-growing HCCs are associated with TGF-β signaling, angiogenesis and inflammation. Our data show that fast HCCs are characterized not only by redundant neo-angiogenesis but also by unique features of distinctively immunosuppressed microenvironment, prominent EMT, and clear-cut activation of TGFβ1 signaling in a general background of long-standing and permanent inflammatory state

Acid-Promoted Competing Pathways in the Oxidative Polymerization of 5,6-Dihydroxyindoles and Related Compounds: Straightforward Cyclotrimerization Routes to Diindolocarbazole Derivatives.

Oxidation of 5,6-dihydroxyindole (la) in acidic aqueous media led to isomeric hexahydroxydiindolocarbazoles, isolated as the acetyl derivatives 5a (29%) and 6a (19%). When the reaction is stopped in the very early stages, small amounts of the indolylindoline 17 and the open trimer 18 can be isolated. Similar oxidation of the N-methyl (1b) and O,O-dimethyl (1c) derivatives of la, as well as of 5-methoxyindole (9b), 6-hydroxyindole (14a), and 6-benzyloxyindole (14b), afforded the corresponding diindolocarbazoles Bb and 6b, 5c and 6c, 10, 16, and the related tetramer 15 in up to 70% overall yield, whereas 5,6-diacetoxyindole (1d), 9a-hydroxyindole (9a), and indole failed to give cyclotrimerization products. Formation of diindolocarbazoles could be explained by a mechanism in which the electron-donating substituents propitiate an array of acid-induced couplings and subsequent dehydrogenation steps driven by the energetically favorable closure of the fused aromatic framework

Virtual prototyping for workplace analysis: the effect of expectation on perceived comfort while using office devices

Computer aided technologies (CAT) are becoming an indispensable instrument to design, improve and manufacture new products and services. Digital human modelling (DHM) systems allow to simulate the Human-artifact interface and to evaluate, in early step of design process, the ergonomic performances of new products or workplaces. In particular, for products that have to be used in a “constrained” workplace, different tasks and activities are associated with different postures. The aim of this study is to investigate the influence of anthropometric characteristics and expectations on the postural comfort perception through the CAT/DHM systems, while using four office devices: desktop computer, laptop computer, tablet and smartphone. A statistical sample of healthy students was selected and their anthropometric characteristics were measured. The postures assumed by the participants were gathered in a not-invasive way by cameras. The angular detection was performed directly on snapshots by using Kinovea ® software. Human joints’ an-gles were used for the virtual-postural analysis, through DELMIA ® software. The evaluation of postural comfort was obtained in two ways: CaMAN ® software developed by the researchers from the Department of Industrial Engineering in Salerno (Italy) was used to calculate the objective comfort indexes while an appropriate questionnaire, given to subjects during the devices usage, was used to evaluate the subjective com-fort indexes. The results of analyses show a difference between subjective and objective postural comfort indexes for all the devices: this difference has been associated to the expectations

The effect of expectation on perceived comfort while using office devices

Different tasks and activities are associated with different sitting postures. The aim of this study is to investigate the influence of anthropometric characteristics and expectations on postural comfort perception while using four office devices: a desktop computer, a laptop computer, a tablet and a smartphone. A statistical sample of healthy students was selected, and their anthropometric characteristics were measured. The postures assumed by the participants were gathered non-invasively using cameras. Angular detection was performed directly on snapshots using Kinovea® software. The angles of human joints were used for the virtualpostural analysis, using DELMIA® software. The evaluation of postural comfort for each of the devices was obtained in two ways: CaMAN® software developed by researchers from the Department of Industrial Engineering in Salerno (Italy) was used to calculate the objective comfort indexes, and a questionnaire was used to evaluate the subjective comfort indexes. The results of the analyses show a difference between subjective and objective postural comfort for all of the devices, and this difference is associated to the expectations

LH and hCG action on the same receptor results in quantitatively and qualitatively different intracellular signalling.

Human luteinizing hormone (hLH) and chorionic gonadotropin (hCG) act on the same receptor (LHCGR) but it is not known whether they elicit the same cellular and molecular response. This study compares for the first time the activation of cell-signalling pathways and gene expression in response to hLH and hCG. Using recombinant hLH and recombinant hCG we evaluated the kinetics of cAMP production in COS-7 and hGL5 cells permanently expressing LHCGR (COS-7/LHCGR, hGL5/LHCGR), as well as cAMP, ERK1/2, AKT activation and progesterone production in primary human granulosa cells (hGLC). The expression of selected target genes was measured in the presence or absence of ERK- or AKT-pathways inhibitors. In COS-7/LHCGR cells, hCG is 5-fold more potent than hLH (cAMP ED(50): 107.1±14.3 pM and 530.0±51.2 pM, respectively). hLH maximal effect was significantly faster (10 minutes by hLH; 1 hour by hCG). In hGLC continuous exposure to equipotent doses of gonadotropins up to 36 hours revealed that intracellular cAMP production is oscillating and significantly higher by hCG versus hLH. Conversely, phospho-ERK1/2 and -AKT activation was more potent and sustained by hLH versus hCG. ERK1/2 and AKT inhibition removed the inhibitory effect on NRG1 (neuregulin) expression by hLH but not by hCG; ERK1/2 inhibition significantly increased hLH- but not hCG-stimulated CYP19A1 (aromatase) expression. We conclude that: i) hCG is more potent on cAMP production, while hLH is more potent on ERK and AKT activation; ii) hGLC respond to equipotent, constant hLH or hCG stimulation with a fluctuating cAMP production and progressive progesterone secretion; and iii) the expression of hLH and hCG target genes partly involves the activation of different pathways depending on the ligand. Therefore, the LHCGR is able to differentiate the activity of hLH and hCG

Dose-response and time-course experiments.

<p><b>a.</b> Dose-response experiment to r-hLH and r-hCG in COS7/LHCGR in the presence of 500 µM IBMX. Total cAMP was measured after 3 hours of incubation and the results of four independent experiments were plotted (Mean±SEM). <b>b.</b> Time-course experiment performed by continuous incubation of COS7/LHCGR for different time-points in the presence of 500 µM IBMX and gonadotropins at ED₅₀ doses (500 pM r-hLH; 100 pM r-hCG). Intracellular cAMP was measured (n = 3; Mean±SEM). Asterisk indicates the significant differences of hLH <i>vs</i> hCG (<i>t-test</i>; <i>p</i><0.05).</p

Primer sequences used in real-time PCR experiments.

*<p>Primer chosen to include all NRG1 transcript variants, verified by NCBI BLAST.</p>**<p>Endogenous control gene.</p