Differential Regulation of White-Opaque Switching by Individual Subunits of Candida Albicans Mediator

The multisubunit eukaryotic Mediator complex integrates diverse positive and negative gene regulatory signals and transmits them to the core transcription machinery. Mutations in individual subunits within the complex can lead to decreased or increased transcription of certain subsets of genes, which are highly specific to the mutated subunit. Recent studies suggest a role for Mediator in epigenetic silencing. Using white-opaque morphological switching in Candida albicans as a model, we have shown that Mediator is required for the stability of both the epigenetic silenced (white) and active (opaque) states of the bistable transcription circuit driven by the master regulator Wor1. Individual deletions of eight C. albicans Mediator subunits have shown that different Mediator subunits have dramatically diverse effects on the directionality, frequency, and environmental induction of epigenetic switching. Among the Mediator deletion mutants analyzed, only Med12 has a steady-state transcriptional effect on the components of the Wor1 circuit that clearly corresponds to its effect on switching. The MED16 and MED9 genes have been found to be among a small subset of genes that are required for the stability of both the white and opaque states. Deletion of the Med3 subunit completely destabilizes the opaque state, even though the Wor1 transcription circuit is intact and can be driven by ectopic expression of Wor1. The highly impaired ability of the med3 deletion mutant to mate, even when Wor1 expression is ectopically induced, reveals that the activation of the Wor1 circuit can be decoupled from the opaque state and one of its primary biological consequences

Particle Shape Characterisation and Classification using Automated Microscopy and Shape Descriptors in Batch Manufacture of Particulate Solids

It is known that size alone, as often defined as the volume equivalent diameter, is not sufficient for characterizing many particulate products. The shape of crystalline products can be as important as size in many applications. Traditionally particulate shape is often defined by some simple descriptors such as the maximum length and aspect ratio. Although these descriptors are intuitive, they result in loss of some information of the original shape. This paper presents a method to use principal component analysis (PCA) to derive simple latent shape descriptors from microscope images of particulate products made in batch processes, and the use of the descriptors for identification of batch to batch variations. Data from batch runs of both a laboratory crystalliser and an industrial crystallisation reactor are analysed using the approach. Qualitative and quantitative comparison with the use of traditional shape descriptors that have physical meanings and Fourier shape descriptors is also made

Incoherent illumination for motion-based imaging through thick scattering medium

Object-motion-based speckle correlation can recover hidden objects from any inhomogeneous medium, which takes advantage of the inherent connection that the cross-correlation between speckle patterns can reflect the autocorrelation of object, providing a route for imaging through or inside thick scattering media. However, once the object is phase-modulated, the above-mentioned relation will not be satisfied under coherent illumination, and the objects cannot be recovered using the existing approaches. Here, we propose an incoherent illumination method for object-motion-based imaging. Theoretical analysis and experimental results show that the cross-correlation between the object-motion-based speckle patterns can be directly used to represent the intensity autocorrelation of the object, making it possible to recover hidden objects regardless of whether the object is phase-modulated or not. Moreover, the proposed approach has a lower root-mean-square error for extracting the autocorrelation patterns of the hidden object. The proposed imaging mechanism blazes a way of imaging moving objects with scattering-induced or intrinsic phase profile, which is in favor of complex imaging scenarios such as inhomogeneous object imaging, deep tissue imaging, and passive lighting scattering imaging

The Tlo Proteins Are Stoichiometric Components of Candida albicans Mediator Anchored via the Med3 Subunit

The amplification of the TLO (for telomere-associated) genes in Candida albicans, compared to its less pathogenic, close relative Candida dubliniensis, suggests a role in virulence. Little, however, is known about the function of the Tlo proteins. We have purified the Mediator coactivator complex from C. albicans (caMediator) and found that Tlo proteins are a stoichiometric component of caMediator. Many members of the Tlo family are expressed, and each is a unique member of caMediator. Protein expression analysis of individual Tlo proteins, as well as the purification of tagged Tlo proteins, demonstrate that there is a large free population of Tlo proteins in addition to the Mediator-associated population. Coexpression and copurification of Tloα12 and caMed3 in Escherichia coli established a direct physical interaction between the two proteins. We have also made a C. albicans med3Δ/Δ strain and purified an intact Mediator from this strain. The analysis of the composition of the med3Δ Mediator shows that it lacks a Tlo subunit. Regarding Mediator function, the med3Δ/Δ strain serves as a substitute for the difficult-to-make tloΔ/Δ C. albicans strain. A potential role of the TLO and MED3 genes in virulence is supported by the inability of the med3Δ/Δ strain to form normal germ tubes. This study of caMediator structure provides initial clues to the mechanism of action of the Tlo genes and a platform for further mechanistic studies of caMediator\u27s involvement in gene regulatory patterns that underlie pathogenesis

Dynamic instabilities in resonant tunneling induced by a magnetic field

We show that the addition of a magnetic field parallel to the current induces self sustained intrinsic current oscillations in an asymmetric double barrier structure. The oscillations are attributed to the nonlinear dynamic coupling of the current to the charge trapped in the well, and the effect of the external field over the local density of states across the system. Our results show that the system bifurcates as the field is increased, and may transit to chaos at large enough fields.Comment: 4 pages, 3 figures, accepted in Phys. Rev. Letter

In-situ crystal morphology identification using imaging analysis with application to the L-glutamic acid crystallization

A synthetic image analysis strategy is proposed for in-situ crystal size measurement and shape identification for monitoring crystallization processes, based on using a real-time imaging system. The proposed method consists of image processing, feature analysis, particle sieving, crystal size measurement, and crystal shape identification. Fundamental image features of crystals are selected for efficient classification. In particular, a novel shape feature, referred to as inner distance descriptor, is introduced to quantitatively describe different crystal shapes, which is relatively independent of the crystal size and its geometric direction in an image captured for analysis. Moreover, a pixel equivalent calibration method based on subpixel edge detection and circle fitting is proposed to measure crystal sizes from the captured images. In addition, a kernel function based method is given to deal with nonlinear correlations between multiple features of crystals, facilitating computation efficiency for real-time shape identification. Case study and experimental results from the cooling crystallization of l-glutamic acid demonstrate that the proposed image analysis method can be effectively used for in-situ crystal size measurement and shape identification with good accuracy

Analysis of Candida albicans Mutants Defective in the Cdk8 Module of Mediator Reveal Links between Metabolism and Biofilm Formation

Candida albicans biofilm formation is a key virulence trait that involves hyphal growth and adhesin expression. Pyocyanin (PYO), a phenazine secreted by Pseudomonas aeruginosa, inhibits both C. albicans biofilm formation and development of wrinkled colonies. Using a genetic screen, we identified two mutants, ssn3Δ/Δ and ssn8Δ/Δ, which continued to wrinkle in the presence of PYO. Ssn8 is a cyclin-like protein and Ssn3 is similar to cyclin-dependent kinases; both proteins are part of the heterotetrameric Cdk8 module that forms a complex with the transcriptional co-regulator, Mediator. Ssn3 kinase activity was also required for PYO sensitivity as a kinase dead mutant maintained a wrinkled colony morphology in the presence of PYO. Furthermore, similar phenotypes were observed in mutants lacking the other two components of the Cdk8 module-Srb8 and Srb9. Through metabolomics analyses and biochemical assays, we showed that a compromised Cdk8 module led to increases in glucose consumption, glycolysis-related transcripts, oxidative metabolism and ATP levels even in the presence of PYO. In the mutant, inhibition of respiration to levels comparable to the PYO-treated wild type inhibited wrinkled colony development. Several lines of evidence suggest that PYO does not act through Cdk8. Lastly, the ssn3 mutant was a hyperbiofilm former, and maintained higher biofilm formation in the presence of PYO than the wild type. Together these data provide novel insights into the role of the Cdk8 module of Mediator in regulation of C. albicans physiology and the links between respiratory activity and both wrinkled colony and biofilm development

Stereo Imaging Camera Model for 3D Shape Reconstruction of Complex Crystals and Estimation of Facet Growth Kinetics

The principle that the 3D shape of crystals that grow from a solution can be characterised in real-time using stereo imaging has been demonstrated previously. It uses the 2D images of a crystal that are obtained from two or more cameras arranged in defined angles as well as a mathematical reconstruction algorithm. Here attention is given to the development of a new and more robust 3D shape reconstruction method for complicated crystal structures. The proposed stereo imaging camera model for 3D crystal shape reconstruction firstly rotates a digitised crystal in the three-dimensional space and varies the size dimensions in all face directions. At each size and orientation, 2D projections of the crystal, according to the angles between the 2D cameras, are recorded. The contour information of the 2D images is processed to calculate Fourier descriptors and radius-based signature that are stored in a database. When the stereo imaging instrument mounted on a crystalliser captures 2D images, the images are segmented to obtain the contour information and processed to obtain Fourier descriptors and radius-based information. The calculated Fourier descriptors and radius-based signature are used to find the best matching in the database. The corresponding 3D crystal shape is thus found. Potash alum crystals that each has 26 habit faces were used as a case study. The result shows that the new approach for 3D shape reconstruction is more accurate and significantly robust than previous methods. In addition, the growth rates of {111}, {110} and {100} faces were correlated with relative supersaturation to derive models of facet growth kinetics