Autophagy: A cyto-protective mechanism which prevents primary human hepatocyte apoptosis during oxidative stress

The role of autophagy in the response of human hepatocytes to oxidative stress remains unknown. Understanding this process may have important implications for the understanding of basic liver epithelial cell biology and the responses of hepatocytes during liver disease. To address this we isolated primary hepatocytes from human liver tissue and exposed them ex vivo to hypoxia and hypoxia-reoxygenation (H-R). We showed that oxidative stress increased hepatocyte autophagy in a reactive oxygen species (ROS) and class III PtdIns3K-dependent manner. Specifically, mitochondrial ROS and NADPH oxidase were found to be key regulators of autophagy. Autophagy involved the upregulation of BECN1, LC3A, Atg7, Atg5 and Atg 12 during hypoxia and H-R. Autophagy was seen to occur within the mitochondria of the hepatocyte and inhibition of autophagy resulted in the lowering a mitochondrial membrane potential and onset of cell death. Autophagic responses were primarily observed in the large peri-venular (PV) hepatocyte subpopulation. Inhibition of autophagy, using 3-methyladenine, increased apoptosis during H-R. Specifically, PV human hepatocytes were more susceptible to apoptosis after inhibition of autophagy. These findings show for the first time that during oxidative stress autophagy serves as a cell survival mechanism for primary human hepatocytes

Skeletal muscle satellite cells cultured in simulated microgravity

Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and therefore provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75 % satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D flat culture and 3-D HARV culture. Plating efficiency (cells attached - cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and non-satellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability since glucose levels in media from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARVS were joined together by cells into three-dimensional aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of satellite cells on microcarrier beads within the HARV bioreactor results in a three dimensional level of organization that could provide a more suitable model to study postnatal muscle development

A bovine model of a respiratory Parachlamydia acanthamoebae infection.

The aim of this study was to evaluate the pathogenicity of Parachlamydia (P.) acanthamoebae as a potential agent of lower respiratory tract disease in a bovine model of induced lung infection. Intrabronchial inoculation with P. acanthamoebae was performed in healthy calves aged 2-3 months using two challenge doses: 10(8) and 10(10) bacteria per animal. Controls received 10(8) heat-inactivated bacteria. Challenge with 10(8) viable Parachlamydia resulted in a mild degree of general indisposition, whereas 10(10) bacteria induced a more severe respiratory illness becoming apparent 1-2 days post inoculation (dpi), affecting 9/9 (100%) animals and lasting for 6 days. The extent of macroscopic pulmonary lesions was as high as 6.6 (6.0)% [median (range)] of lung tissue at 2-4 dpi and correlated with parachlamydial genomic copy numbers detected by PCR, and with bacterial load estimated by immunohistochemistry in lung tissue. Clinical outcome, acute phase reactants, pathological findings and bacterial load exhibited an initial dose-dependent effect on severity. Animals fully recovered from clinical signs of respiratory disease within 5 days. The bovine lung was shown to be moderately susceptible to P. acanthamoebae, exhibiting a transient pneumonic inflammation after intrabronchial challenge. Further studies are warranted to determine the precise pathophysiologic pathways of host-pathogen interaction

Evaluation of saliency tracking as an alternative for health monitoring in PMSM-drives under nonstationary conditions

This paper evaluates the capability of saliency tracking to assess the health condition of permanent magnet synchronous motor (PMSM) drives operating under nonstationary conditions. The evaluated scheme is based on saliency tracking methods, which are associated to the accurate sensorless control of AC drives without zero speed limitations. In this work two representative saliency tracking architectures are evaluated: High Frequency (HF) injection, and PWM transient excitation. Although a monitoring approach based on HF injection was previously reported, a comparative study to evaluate the most representative saliency tracking schemes to assess health condition in drives was still missing. The aim of this work is to fill out this gap by evaluating and comparing two saliency-based monitoring schemes (one based on HF-injection and the other based on PWM transient excitation) to evaluate their performance in the presence of inter-turn winding faults. Simulation and experimental results are presented which confirm that both schemes offer excellent detection capabilities and that are suitable for drives operating under nonstationary conditions including standstill operation. Significant differences are also found for instance, PWM transient excitation offers improved accuracy since the approach is not affected by the inverter nonlinearities and is suitable for full-speed range applications. The main drawback here is complexity and the hardware requirements. Schemes based on HF-injection proved to be very simple and provide comparable results; however a good performance is only guaranteed for the zero-to-medium speed range applications which limit their applicability

Condition monitoring approach for permanent magnet synchronous motor drives based on the INFORM method

This paper proposes a monitoring scheme based on saliency tracking to assess the health condition of PMSM drives operating under non stationary conditions. The evaluated scheme is based on the INFORM methodology, which is associated to the accurate sensorless control of PM drives without zero speed limitation. The result is a monitoring scheme that is able to detect faults that would be very difficult to evaluate under nonstationary conditions. A relevant aspect of the proposed scheme is that it remains valid for full speed range, and can be used for standstill operation. Additionally, the approach is insensitive to the inverter nonlinearities which enhance the detection capabilities further respect to similar topologies. In this work the proposed approach is evaluated numerically and experimentally in the presence of incipient winding faults and inter-turn short circuits in a PM conventional drive. The obtained results show quick response and excellent detection capabilities not only in the detection of faults, but to determine their magnitude which is vital to avoid further degradation