Effective control of Leptosphaeria maculans increases importance of L. biglobosa as a cause of phoma stem canker epidemics on oilseed rape

BACKGROUND: Phoma stem canker is a damaging disease of oilseed rape caused by two related fungal species, Leptosphaeria maculans and L. biglobosa. However, previous work has mainly focused on L. maculans and there has been little work on L. biglobosa. This work provides evidence of the importance of L. biglobosa to stem canker epidemics in the UK. RESULTS: Quantification of L. maculans and L. biglobosa DNA using species-specific quantitative PCR showed that L. biglobosa caused both upper stem lesions and stem base cankers on nine oilseed rape cultivars in the UK. Upper stem lesions were mainly caused by L. biglobosa. For stem base cankers, there was more L. maculans DNA than L. biglobosa DNA in the susceptible cultivar Drakkar, while there was more L. biglobosa DNA than L. maculans DNA in cultivars with the resistance gene Rlm7 against L. maculans. The frequency of L. biglobosa detected in stem base cankers increased from 14% in 2000 to 95% in 2013. Ascospores of L. biglobosa and L. maculans were mostly released on the same days and the number of L. biglobosa ascospores in air samples increased from the 2010/2011 to 2012/2013 growing seasons. CONCLUSION: Effective control of L. maculans increased infection by L. biglobosa, causing severe upper stem lesions and stem base cankers, leading to yield losses. The importance of L. biglobosa to phoma stem canker epidemics needs to target both L. maculans and L. biglobosa

Structure-function studies of the bHLH phosphorylation domain of TWIST1 in prostate cancer cells

The TWIST1 gene has diverse roles in development and pathologic diseases such as cancer. TWIST1 is a dimeric basic helix-loop-helix (bHLH) transcription factor existing as TWIST1-TWIST1 or TWIST1-E12/47. TWIST1 partner choice and DNA binding can be influenced during development by phosphorylation of Thr125 and Ser127 of the Thr-Gln-Ser (TQS) motif within the bHLH of TWIST1. The significance of these TWIST1 phosphorylation sites for metastasis is unknown. We created stable isogenic prostate cancer cell lines overexpressing TWIST1 wild-type, phospho-mutants, and tethered versions. We assessed these isogenic lines using assays that mimic stages of cancer metastasis. In vitro assays suggested the phospho-mimetic Twist1-DQD mutation could confer cellular properties associated with pro-metastatic behavior. The hypo-phosphorylation mimic Twist1-AQA mutation displayed reduced pro-metastatic activity compared to wild-type TWIST1 in vitro, suggesting that phosphorylation of the TWIST1 TQS motif was necessary for pro-metastatic functions. In vivo analysis demonstrates that the Twist1-AQA mutation exhibits reduced capacity to contribute to metastasis, whereas the expression of the Twist1-DQD mutation exhibits proficient metastatic potential. Tethered TWIST1-E12 heterodimers phenocopied the Twist1-DQD mutation for many in vitro assays, suggesting that TWIST1 phosphorylation may result in heterodimerization in prostate cancer cells. Lastly, the dual phosphatidylinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) inhibitor BEZ235 strongly attenuated TWIST1-induced migration that was dependent on the TQS motif. TWIST1 TQS phosphorylation state determines the intensity of TWIST1-induced pro-metastatic ability in prostate cancer cells, which may be partly explained mechanistically by TWIST1 dimeric partner choice

High Strength Thermoplastic Bonding for Multi-channel, Multi-layer Lab-on-Chip Devices for Ocean and Environmental applications

A solvent vapour thermoplastic bonding process is reported which provides high-strength bonding of PMMA over a large area for multi-channel and multi-layer microfluidic devices with shallow high-resolution channel features. The bond process utilises a low-temperature vacuum thermal fusion step with prior exposure of the substrate to chloroform (CHCl3) vapour to reduce bond temperature to below the PMMA glass transition temperature. Peak tensile and shear bond strengths > 3 MPa were achieved for a typical channel depth reduction of 25 A mu m. The device-equivalent bond performance was evaluated for multiple layers and high-resolution channel features using double-side and single-side exposure of the bonding pieces. A single-sided exposure process was achieved which is suited to multi-layer bonding with channel alignment at the expense of greater depth loss and a reduction in peak bond strength. However, leak and burst tests demonstrate bond integrity up to at least 10 bar channel pressure over the full substrate area of 100 mm x 100 mm. The inclusion of metal tracks within the bond resulted in no loss of performance. The vertical wall integrity between channels was found to be compromised by solvent permeation for wall thicknesses of 100 A mu m which has implications for high-resolution serpentine structures. Bond strength is reduced considerably for multi-layer patterned substrates where features on each layer are not aligned, despite the presence of an intermediate blank substrate. Overall a high-performance bond process has been developed that has the potential to meet the stringent specifications for lab-on-chip deployment in harsh environmental conditions for applications such as deep ocean profiling

A Synthetic Dolastatin 10 Analogue Suppresses Microtubule Dynamics, Inhibits Cell Proliferation, and Induces Apoptotic Cell Death

We have synthesized eight analogues (D1-D8) of dolastatin 10 containing several unique amino acid subunits. Of these agents, D5 was found to be most effective in inhibiting both He La cell proliferation and microtubule assembly in vitro. At low nanomolar concentrations, 135 inhibited the proliferation of several types of cancer cells in culture. D5 bound to tubulin with a dissociation constant of 29.4 +/- 6 mu M. D5 depolymerized microtubules in cultured cells and produced mulitpolar spindles. At its half-maximal inhibitory concentration (15 nM), DS strongly suppressed the dynamics of individual microtubules in live MCF-7 cells. DS increased the accumulation of checkpoint proteins BubR1 and Mad2 at the kinetochoric region and caused G2/M block in these cells. The blocked cells underwent apoptosis with the activation of Jun N-terminal kinase. The results suggested that DS exerts its antiproliferative action by dampening microtubule dynamics

Development of Smart Number Writing Robotic Arm using Stochastic Gradient Decent Algorithm

Robotics and Neural Networks will play a major role in the future of manufacturing and automation process. Nowadays not many robotic systems are smart systems, in the sense that they operate on a predefined algorithm to do their task. This research focuses on a design and development of a robotic arm with a visual input. The robotic arm will perform its job with the help of visual aid. The system will analyze the input image upon which the decision to write a number using Stochastic Gradient Decent (SGD) algorithm. In a nutshell this research work shows how the neural network can be incorporated with robot arm control, which is a desired field of interest in development of smart robotic systems. This work presents where the robotic arm is incorporated together with a neural network to perform a task of writing numbers using vision

In-Silico Skin Model: A Multiscale Simulation Study of Drug Transport

Accurate in-silico models are required to predict the release of drug molecules through skin in order to supplement the in-vivo experiments for faster development/testing of drugs. The upper most layer of the skin, stratum corneum (SC), offers the main resistance for permeation of actives. Most of the SC’s molecular level models comprise cholesterol and phospholipids only, which is far from reality. In this study we have implemented a multiscale modeling framework to obtain the release profile of three drugs, namely, caffeine, fentanyl, and naphthol, through skin SC. We report for the first time diffusion of drugs through a realistic skin molecular model comprised of ceramides, cholesterol, and free fatty acid. The diffusion coefficients of drugs in the SC lipid matrix were determined from multiple constrained molecular dynamics simulations. The calculated diffusion coefficients were then used in the macroscopic models to predict the release profiles of drugs through the SC. The obtained release profiles were in good agreement with available experimental data. The partition coefficient exhibits a greater effect on the release profiles. The reported multiscale modeling framework would provide insight into the delivery mechanisms of the drugs through the skin and shall act as a guiding tool in performing targeted experiments to come up with a suitable delivery system