Autonomous growth in serum-free medium and production of hepatocellular carcinomas by differentiated hepatocyte lines that overexpress transforming growth factor alpha 1

Transforming growth factor alpha (TGF-alpha) is a polypeptide closely associated with hepatocyte proliferation in vivo and in vitro. In order to investigate the mechanisms by which TGF-alpha contributes to hepatocyte replication and transformation, we isolated hepatocytes from mice bearing a human TGF-alpha transgene and examined their growth properties and gene expression in defined, serum-free culture. The transgenic hepatocytes continued to overexpress human TGF-alpha mRNA and peptide, and were able to proliferate without exogenous growth factors in primary culture, in contrast to nontransgenic mouse hepatocytes. In short-term culture the transgenic hepatocytes underwent 1 wave of DNA replication at 72-96 h in culture before senescing, similar to nontransgenic hepatocytes supplemented with epidermal growth factor. Constitutive expression of TGF-alpha rendered the transgenic hepatocytes unresponsive to further growth stimulation by exogenous TGF-alpha, as well as other mitogens such as epidermal growth factor and hepatocyte growth factor. However, it did not alter their sensitivity to growth inhibition by TGF beta 1, 2 and 3. The addition of nicotinamide to the culture medium enabled both transgenic and epidermal growth factor-supplemented normal hepatocytes to replicate repeatedly and survive for > or = 2 months in primary culture while maintaining differentiated traits. From these long-term primary cultures of transgenic and nontransgenic hepatocytes, we established immortalized cell lines (designated TAMH and NMH lines, respectively). Both lines continued to express differentiated adult hepatocytic markers such as albumin, alpha-1-antitrypsin, transferrin, and connexin 26 and 32 mRNAs, but also expressed mRNAs for the oncofetal markers alpha-fetoprotein and insulin-like growth factor II. Unlike the near-diploid NMH hepatocyte line, the transgenic TAMH hepatocyte line was quasi-tetraploid, strongly expressed human TGF-alpha mRNA, and was highly tumorigenic in nude mice. Well-differentiated hepatocellular carcinomas developed in nude mice given injections of the TAMH line, and these appeared similar to the primary liver tumors seen in TGF-alpha transgenic mice with regard to histology and strong expression of mouse and human TGF-alpha, insulin-like growth factor II, and alpha-fetoprotein mRNAs. Our data show that TGF-alpha overexpression causes autonomous hepatocyte proliferation and contributes to neoplasia but that additional cellular alterations must occur for carcinogenesis. Inappropriate expression of insulin-like growth factor II may constitute one of these steps. The TGF-alpha transgenic mouse hepatocyte line TAMH appears to undergo transformation in a similar manner to that of hepatocytes overexpressing TGF-alpha in vivo, and should serve as an ideal system in which to study hepatocarcinogenesis.     

Modelling the Floating Ladder Track Response to a Moving Load by an Infinite Bernoulli-Euler Beam on Periodic Flexible Supports

An infinite Bernoulli-Euler beam (representing the "combined rail" consisting of the rail and longitudinal sleeper) mounted on periodic flexible point supports (representing the railpads) has already proven to be a suitable mathematical model for the floating ladder track (FLT), to define its natural vibrations and its forced response due to a moving load. Adopting deliberately conservative parameters for the existing FLT design, we present further results for the response to a steadily (uniformly) moving load when the periodic supports are assumed to be elastic, and then introduce the mass and viscous damping of the periodic supports. Typical support damping significantly moderates the resulting steady deflexion at any load speed, and in particular substantially reduces the magnitude of the resonant response at the critical speed. The linear mathematical analysis is then extended to include the inertia of the load that otherwise moves uniformly along the beam, generating overstability at supercritical speeds — i.e. at load speeds notably above the critical speed predicted for the resonant response when the load inertia is neglected. Neither the resonance nor the overstability should prevent the safe implementation of the FLT design in modern high speed rail systems.     

Thermoeconomic impact on combined cycle performance of advanced blade cooling systems

In this work the thermoeconomic features of two different combined cycles using air “open loop” and steam “closed loop” cooled gas turbines are presented and compared in depth. In order to properly estimate both thermodynamic and thermoeconomic performance of the different combined cycles an analytical model of the blade cooling system has been developed in details and outlined in the paper. Internal Thermoeconomic functional analysis is not performed here, as only economic results are shown and discussed. The blade cooling detailed model, originally developed by TPG researchers, has been integrated into the web based modular code WTEMP, already validated for GT based cycles, developed in the last ten years by TPG. It is shown that the closed loop blade cooling configuration has the greatest potential in terms of thermodynamic efficiency and economic competitivity in the mid-term.     

Ethanol steam reforming kinetics of a Pd–Ag membrane reactor

The ethanol steam reforming reaction carried out in a Pd-based tubular membrane reactor has been modelled via a finite element code. The model considers the membrane tube divided into finite volume elements where the mass balances for both lumen and shell sides are carried out accordingly to the reaction and permeation kinetics. Especially, a simplified “power law” has been applied for the reaction kinetics: the comparison with experimental data obtained by using three different kinds of catalyst (Ru, Pt and Ni based) permitted defining the coefficients of the kinetics expression as well as to validate the model. Based on the Damkohler–Peclet analysis, the optimization of the membrane reformer has been also approached.     

Differential Electrochemical Conductance Imaging at the Nanoscale

Electron transfer in proteins is essential in crucial biological processes. Although the fundamental aspects of biological electron transfer are well characterized, currently there are no experimental tools to determine the atomic‐scale electronic pathways in redox proteins, and thus to fully understand their outstanding efficiency and environmental adaptability. This knowledge is also required to design and optimize biomolecular electronic devices. In order to measure the local conductance of an electrode surface immersed in an electrolyte, this study builds upon the current–potential spectroscopic capacity of electrochemical scanning tunneling microscopy, by adding an alternating current modulation technique. With this setup, spatially resolved, differential electrochemical conductance images under bipotentiostatic control are recorded. Differential electrochemical conductance imaging allows visualizing the reversible oxidation of an iron electrode in borate buffer and individual azurin proteins immobilized on atomically flat gold surfaces. In particular, this method reveals submolecular regions with high conductance within the protein. The direct observation of nanoscale conduction pathways in redox proteins and complexes enables important advances in biochemistry and bionanotechnology.     

A digital filter-based approach to the remote condition monitoring of railway turnouts

Railway operations in Europe have changed dramatically since the early 1990s, partly as a result of new European Union Directives. Performance targets have become more and more exacting, due to reductions in state support for railways and the need to increasing traffic. More intensive operations also place greater demands on the hardware of the railway. This is true for both rolling stock and infrastructure subsystems and components, particularly so in the case of the latter where the time available for maintenance is being reduced. The authors of this paper focus on the railway infrastructure, and more specifically on points. These are critical elements whose reliability is key to the operation of the whole system. Using intelligent monitoring systems, it is possible to predict problems and enable quick recovery before component failures disrupt operations. The authors have studied the application of remote condition monitoring to point mechanisms and their operation, and have identified algorithms which may be used to identify incipient failures. In this paper, the authors propose a Kalman filter for the linear discrete data filtering problem encountered when using current sensor data in a point condition monitoring system. The reason for applying Kalman filtering in this study was to increase the reliability of the model presented to the rule-based decision mechanism.     

{\alpha^{\prime}}_{\text{H}} -Dicalcium silicate bone cement doped with tricalcium phosphate: characterization,bioactivity and biocompatibility

The influence of phosphorus doping on the properties of $ \alpha^{\prime}_{\text{H}} $ -dicalcium silicate (C₂S) bone cement was analyzed, in addition to bioactivity and biocompatibility. All the cements were composed of a solid solution of TCP in C₂S ( $ \alpha^{\prime}_{\text{H}} $ -C₂S_ss) as the only phase present. The compressive strength ranged from 3.8–16.3 MPa. Final setting times ranged from 10 to 50 min and were lower for cements with lower L/P content. Calcium silicate hydrate was the principal phase formed during the hydration process of the cements. The cement exhibited a moderate degradation and could induce carbonated hydroxyapatite formation on its surface and into the pores. The cell attachment test showed that the $ \alpha^{\prime}_{\text{H}} $ -Ca₂SiO₄ solid solution supported human adipose stem cells adhesion and spreading, and the cells established close contacts with the cement after 24 h of culture. The novel $ \alpha^{\prime}_{\text{H}} $ -C₂S_ss cements might be suitable for potential applications in the biomedical field, preferentially as materials for bone/dental repair.     

Analysis of liver regeneration in mice lacking type 1 or type 2 tumor necrosis factor receptor: requirement for type 1 but not type 2 receptor

We used KO mice lacking either TNF receptor 1 (TNFR-1) or receptor 2 (TNFR-2) to determine whether signaling at the start of liver regeneration after partial hepatectomy (PH) involves only one or both TNF receptors and to analyze in more detail the abnormalities caused by lack of TNFR-1 receptor, which is required for the initiation of liver regeneration. Lack of TNFR-2 had little effect on NF-kappaB and STAT3 binding, and no effect in interleukin-6 production after PH, but caused a delay in AP-1 and C/EBP binding and in the expression of c-jun and c-myc messenger RNA (mRNA). In contrast to mice lacking TNFR-1, which had deficient hepatocyte DNA synthesis and massive lipid accumulation in hepatocytes, TNFR-2 KO mice had normal liver structure and similar levels of hepatocyte DNA replication as those of wild type mice. We conclude that TNFR-1, but not TNFR-2, is necessary for liver regeneration, and that NF-kappaB and STAT3 binding are activated by signals transduced by TNFR-1. Inhibition of AP-1 and C/EBP binding and in the expression of c-jun and c-myc mRNA in the first 4 hours after PH, as well as the apparent lack of Fos in AP-1 complexes, had no effect on the timing or extent of DNA replication.