Increasing leucine concentration stimulates mechanistic target of rapamycin signaling and cell growth in C2C12 skeletal muscle cells

Leucine is a key amino acid for initiating translation in muscle cells, but the dose-dependent effects of leucine on intracellular signaling are poorly characterized. This study examined the effect that increasing doses of leucine would have on changes in mechanistic target of rapamycin (mTOR)–mediated signaling, rates of protein synthesis, and cell size in C2C12 cells. We hypothesized that a leucine “threshold” exists, which represents the minimum stimulus required to initiate mTOR signaling in muscle cells. Acute exposure to 1.5, 3.2, 5.0, and 16.1 mM leucine increased phosphorylation of mTORSer2448 (~1.4-fold; P < .04), 4E-BP1 Thr37/46 (~1.9-fold; P < .001), and rpS6Ser235/6 (~2.3-fold; P < .001). However, only p70S6kThr389 exhibited a dose-dependent response to leucine with all treatments higher than control (~4-fold; P < .001) and at least 5 mM higher than the 1.5-mM concentration (1.2-fold; P < .02). Rates of protein synthesis were not altered by any treatment. Seven days of exposure to 0.5, 1.5, 5.0, and 16.5 mM leucine resulted in an increase in cell size in at least 5 mM treatments (~1.6-fold, P < .001 vs control). Our findings indicate that even at low leucine concentrations, phosphorylation of proteins regulating translation initiation signaling is enhanced. The phosphorylation of p70S6kThr389 follows a leucine dose-response relationship, although this was not reflected by the acute protein synthetic response. Nevertheless, under the conditions of the present study, it appears that leucine concentrations of at least 5 mM are necessary to enhance cell growth.

Board monitoring and the balanced scorecard in nonprofits : challenges in realising potential

Directors of nonprofits in most countries have legal responsibility for monitoring organisational performance (Brody 2010), although there is typically little guidance on how this should occur. The balanced scorecard (BSC) (Kaplan & Norton, 1996, 2001) potentially provides boards with a monitoring tool (Kaplan $ Norton, 2006; Lorsch, 2002). The BSC is intended to help integrate performance measurement, performance management and strategy implmentation (Kaplan 2009). The scorecards is balanced in that it should incorporate both financial and non-financial measures, external and internal perspectives, short and long-term objectives and both lagging and leading indicators. It is a relatively simple tool, but with potentially profound implications for directing board attention and sbusequent action (Ocasio, 1997; Salterio, 2012).

Identifying weak linear features with the "coalescing shortest path image transform"

The detection of line-like features in images finds many applications in microanalysis. Actin fibers, microtubules, neurites, pilis, DNA, and other biological structures all come up as tenuous curved lines in microscopy images. A reliable tracing method that preserves the integrity and details of these structures is particularly important for quantitative analyses. We have developed a new image transform called the "Coalescing Shortest Path Image Transform" with very encouraging properties. Our scheme efficiently combines information from an extensive collection of shortest paths in the image to delineate even very weak linear features. © Copyright Microscopy Society of America 2011.

Tubule detection in testis images using boundary weighting and circular shortest path

In studies of germ cell transplantation, measureing tubule diameters and counting cells from different populations using antibodies as markers are very important. Manual measurement of tubule sizes and cell counts is a tedious and sanity grinding work. In this paper, we propose a new boundary weighting based tubule detection method. We first enhance the linear features of the input image and detect the approximate centers of tubules. Next, a boundary weighting transform is applied to the polar transformed image of each tubule region and a circular shortest path is used for the boundary detection. Then, ellipse fitting is carried out for tubule selection and measurement. The algorithm has been tested on a dataset consisting of 20 images, each having about 20 tubules. Experiments show that the detection results of our algorithm are very close to the results obtained manually. © 2013 IEEE.

Microalgal growth characteristics and subsequent influence on dewatering efficiency

Dewatering of microalgal culture is a major bottleneck towards the industrial-scale processing of microalgae for bio-diesel production. The dilute nature of harvested microalgal cultures poses a huge operation cost to dewater; thereby rendering microalgae-based fuels less economically attractive. This study explores the influence of microalgal growth phases and intercellular interactions during cultivation on dewatering efficiency of microalgae cultures. Experimental results show that microalgal cultures harvested during a low growth rate phase (LGRP) of 0.03 d-1 allowed a higher rate of settling than those harvested during a high growth rate phase (HGRP) of 0.11 d-1, even though the latter displayed a higher average differential biomass concentration of 0.2 g L-1 d-1. Zeta potential profile during the cultivation process showed a maximum electronegative value of -43.2 ± 0.7 mV during the HGRP which declined to stabilization at -34.5 ± 0.4 mV in the LGRP. The lower settling rate observed for HGRP microalgae is hence attributed to the high stability of the microalgal cells which electrostatically repel each other during this growth phase. Tangential flow filtration of 20 L HGRP culture concentrated 23 times by consuming 0.51 kWh/m3 of supernatant removed whilst 0.38 kWh/m3 was consumed to concentrate 20 L of LGRP by 48 times.

Growth medium selection and its economic impact on plasmid DNA production

Current developments in gene medicine and vaccination studies are utilizing plasmid DNA (pDNA) as the vector. For this reason, there has been an increasing trend towards larger and larger doses of pDNA utilized in human trials: from 100-1000 μg in 2002 to 500-5000 μg in 2005. The increasing demand of pDNA has created the need to revolutionalize current production levels under optimum economy. In this work, different standard media (LB, TB and SOC) for culturing recombinant Escherichia coli DH5α harbouring pUC19 were compared to a medium optimised for pDNA production. Lab scale fermentations using the standard media showed that the highest pDNA volumetric and specific yields were for TB (11.4 μg/ml and 6.3 μg/mg dry cell mass respectively) and the lowest was for LB (2.8 μg/ml and 3.3 μg/mg dry cell mass respectively). A fourth medium, PDMR, designed by modifying a stoichiometrically-formulated medium with an optimised carbon source concentration and carbon to nitrogen ratio displayed pDNA volumetric and specific yields of 23.8 μg/ml and 11.2 μg/mg dry cell mass respectively. However, it is the economic advantages of the optimised medium that makes it so attractive. Keeping all variables constant except medium and using LB as a base scenario (100 medium cost [MC] units/mg pDNA), the optimised PDMR medium yielded pDNA at a cost of only 27 MC units/mg pDNA. These results show that greater amounts of pDNA can be obtained more economically with minimal extra effort simply by using a medium optimised for pDNA production.

Perturbed Lignification Impacts Tree Growth in Hybrid Poplar-A Function of Sink Strength, Vascular Integrity, and Photosynthetic Assimilation

The effects of reductions in cell wall lignin content, manifested by RNA interference suppression of coumaroyl 3'-hydroxylase, on plant growth, water transport, gas exchange, and photosynthesis were evaluated in hybrid poplar trees (Populus alba 3 grandidentata). The growth characteristics of the reduced lignin trees were significantly impaired, resulting in smaller stems and reduced root biomass when compared to wild-type trees, as well as altered leaf morphology and architecture. The severe inhibition of cell wall lignification produced trees with a collapsed xylem phenotype, resulting in compromised vascular integrity, and displayed reduced hydraulic conductivity and a greater susceptibility to wall failure and cavitation. In the reduced lignin trees, photosynthetic carbon assimilation and stomatal conductance were also greatly reduced, however, shoot xylem pressure potential and carbon isotope discrimination were higher and water-use efficiency was lower, inconsistent with water stress. Reductions in assimilation rate could not be ascribed to increased stomatal limitation. Starch and soluble sugars analysis of leaves revealed that photosynthate was accumulating to high levels, suggesting that the trees with substantially reduced cell wall lignin were not carbon limited and that reductions in sink strength were, instead, limiting photosynthesis.

Convergence-guaranteed time-varying RRT path planning for profiling floats in 4-Dimensional flow

This paper presents an extension to the Rapidly-exploring Random Tree (RRT) algorithm applied to autonomous, drifting underwater vehicles. The proposed algorithm is able to plan paths that guarantee convergence in the presence of time-varying ocean dynamics. The method utilizes 4-Dimensional, ocean model prediction data as an evolving basis for expanding the tree from the start location to the goal. The performance of the proposed method is validated through Monte-Carlo simulations. Results illustrate the importance of the temporal variance in path execution, and demonstrate the convergence guarantee of the proposed methods.

Practical path planning and obstacle avoidance for autonomous mowing

There is a need for systems which can autonomously perform coverage tasks on large outdoor areas. Unfortunately, the state-of-the-art is to use GPS based localization, which is not suitable for precise operations near trees and other obstructions. In this paper we present a robotic platform for autonomous coverage tasks. The system architecture integrates laser based localization and mapping using the Atlas Framework with Rapidly-Exploring Random Trees path planning and Virtual Force Field obstacle avoidance. We demonstrate the performance of the system in simulation as well as with real world experiments.

Regional aviation and economic growth : cointegration and causality analysis in Australia

Air transport is a critical link to regional, rural and remote communities in Australia. Air services provide important economic and social benefits but very little research has been done on assessing the value of regional aviation. This research provides the first empirical evidence that there is short and long run causality between regional aviation and economic growth. The authors analysed 88 regional airports in Australia over a period of 1985–86 to 2010–11 to determine the catalytic impacts of regional air transport on regional economic growth. The analysis was conducted using annual data related to total airport passenger movements – for the level of airport activity, and real aggregate taxable income – to represent economic growth. A significant bi-directional relationship was established: airports have an impact on regional economic growth and the economy directly impacts regional air transport. The economic significance of regional air transport confirms the importance of the airport as infrastructure for regional councils and the need for them to maintain and develop local airports. Funding should be targeted at airports directly to support regional development.