Temperature-Sensitive Src-Transformed Mdck Cells nn 3d Cultures as a Model of Malignant Transformation of Epithelial Cells

The equilibrium between cell proliferation, differentiation, and apoptosis is crucial for maintaining homeostasis in epithelial tissues. In order for the epithelium to function properly, individual cells must gain normal structural and functional polarity. The junctional proteins have an important role both in binding the cells together and in taking part in cell signaling. Cadherins form adherens junctions. Cadherins initiate the polarization process by first recognizing and binding the neighboring cells together, and then guiding the formation of tight junctions. Tight junctions form a barrier in dividing the plasma membranes to apical and basolateral membrane domains. In glandular tissues, single layered and polarized epithelium is folded into tubes or spheres, in which the basal side of the epithelial layer faces the outer basal membrane, and the apical side the lumen. In carcinogenesis, the differentiated architecture of an epithelial layer is disrupted. Filling of the luminal space is a hallmark of early epithelial tumors in tubular and glandular structures. In order for the transformed tumor cells to populate the lumen, enhanced proliferation as well as inhibition of apoptosis is required. Most advances in cancer biology have been achieved by using two-dimensional (2D) cell culture models, in which the cells are cultured on flat surfaces as monolayers. However, the 2D cultures are limited in their capacity to recapitulate the structural and functional features of tubular structures and to represent cell growth and differentiation in vivo. The development of three-dimensional (3D) cell culture methods enables the cells to grow and to be studied in a more natural environment. Despite the wide use of 2D cell culture models and the development of novel 3D culture methods, it is not clear how the change of the dimensionality of culture conditions alters the polarization and transformation process and the molecular mechanisms behind them. Src is a well-known oncogene. It is found in focal and adherens junctions of cultured cells. Active src disrupts cell-cell junctions and interferes with cell-matrix binding. It promotes cell motility and survival. Src transformation in 2D disrupts adherens junctions and the fibroblastic phenotype of the cells. In 3D, the adherens junctions are weakened, and in glandular structures, the lumen is filled with nonpolarized vital cells. Madin-Darby canine kidney (MDCK) cells are an epithelial cell type commonly used as a model for cell polarization. Its-src-transformed variants are useful model systems for analyzing the changes in cell morphology, and they play a role in src-induced malignant transformation. This study investigates src-transformed cells in 3D cell cultures as a model for malignant transformation. The following questions were posed. Firstly: What is the role of the composition and stiffness of the extracellular matrix (ECM) on the polarization and transformation of ts v-src MDCK cells in 3D cell cultures? Secondly: How do the culture conditions affect gene expression? What is the effect of v-src transformation in 2D and in 3D cell models? How does the shift from 2D to 3D affect cell polarity and gene expression? Thirdly: What is the role of survivin and its regulator phosphatase and tensin homolog protein (PTEN) in cell polarization and transformation, and in determining cell fate? How does their expression correlate with impaired mitochondrial function in transformed cells? In order to answer the above questions, novel methods of culturing and monitoring cells had to be created: novel 3D methods of culturing epithelial cells were engineered, enabling real time monitoring of a polarization and transformation process, and functional testing of 3D cell cultures. Novel 3D cell culture models and imaging techniques were created for the study. Attention was focused especially on confocal microscopy and live-cell imaging. Src-transformation disturbed the polarization of the epithelium by disrupting cell adhesion, and sensitized the cells to their environment. With active src, the morphology of the cell cluster depended on the composition and stiffness of the matrix. Gene expression studies revealed a broader impact of src transformation than mere continuous activity of src-kinase. In 2D cultures, src transformation altered the expression of immunological, actin cytoskeleton and extracellular matrix (ECM). In 3D, the genes regulating cell division, inhibition of apoptosis, cell metabolism, mitochondrial function, actin cytoskeleton and mechano-sensing proteins were altered. Surprisingly, changing the culture conditions from 2D to 3D affected also gene expression considerably. The microarray hit survivin, an inhibitor of apoptosis, played a crucial role in the survival and proliferation of src-transformed cells.

A model for heat transfer in a honey bee swarm

A swarm is a temporary structure formed when several thousand honey bees leave their hive and settle on some object such as the branch of a tree. They remain in this position until a suitable site for a new home is located by the scout bees. A continuum model based on heat conduction and heat generation is used to predict temperature profiles in swarms. Since internal convection is neglected, the model is applicable only at low values of the ambient temperature T-a. Guided by the experimental observations of Heinrich (1981a-c, J. Exp. Biol. 91, 25-55; Science 212, 565-566; Sci. Am. 244, 147-160), the analysis is carried out mainly for non-spherical swarms. The effective thermal conductivity is estimated using the data of Heinrich (1981a, J. Exp. Biol. 91, 25-55) for dead bees. For T-a = 5 and 9 degrees C, results based on a modified version of the heat generation function due to Southwick (1991, The Behaviour and Physiology of Bees, PP 28-47. C.A.B. International, London) are in reasonable agreement with measurements. Results obtained with the heat generation function of Myerscough (1993, J. Theor. Biol. 162, 381-393) are qualitatively similar to those obtained with Southwick's function, but the error is more in the former case. The results suggest that the bees near the periphery generate more heat than those near the core, in accord with the conjecture of Heinrich (1981c, Sci. Am. 244, 147-160). On the other hand, for T-a = 5 degrees C, the heat generation function of Omholt and Lonvik (1986, J. Theor. Biol. 120, 447-456) leads to a trivial steady state where the entire swarm is at the ambient temperature. Therefore an acceptable heat generation function must result in a steady state which is both non-trivial and stable with respect to small perturbations. Omholt and Lonvik's function satisfies the first requirement, but not the second. For T-a = 15 degrees C, there is a considerable difference between predicted and measured values, probably due to the neglect of internal convection in the model.

Voltage and Temperature-Aware SSTA Using Neural Network Delay Model

With the emergence of voltage scaling as one of the most powerful power reduction techniques, it has been important to support voltage scalable statistical static timing analysis (SSTA) in deep submicrometer process nodes. In this paper, we propose a single delay model of logic gate using neural network which comprehensively captures process, voltage, and temperature variation along with input slew and output load. The number of simulation programs with integrated circuit emphasis (SPICE) required to create this model over a large voltage and temperature range is found to be modest and 4x less than that required for a conventional table-based approach with comparable accuracy. We show how the model can be used to derive sensitivities required for linear SSTA for an arbitrary voltage and temperature. Our experimentation on ISCAS 85 benchmarks across a voltage range of 0.9-1.1V shows that the average error in mean delay is less than 1.08% and average error in standard deviation is less than 2.85%. The errors in predicting the 99% and 1% probability point are 1.31% and 1%, respectively, with respect to SPICE. The two potential applications of voltage-aware SSTA have been presented, i.e., one for improving the accuracy of timing analysis by considering instance-specific voltage drops in power grids and the other for determining optimum supply voltage for target yield for dynamic voltage scaling applications.

A Model for the Temperature Distribution in Resin Impregnated Paper Bushings

Resin impregnated paper (RIP) is a relatively new insulation system recommended for the use in transformer bushings. In the recent past, RIP has acquired prominence as insulation in bushings, over conventional oil impregnated paper (OIP), in view of its overwhelming advantages the more important among them being low dielectric loss and possibility for positioning the bushing at any desired angle over the transformer. In addition, the fact that such systems do not pose problems of fire hazard is counted as a very important consideration. The disadvantage of RIP compared to OIP, however, is its much higher cost and involved manufacturing process. The temperature rise in RIP bushings under normal operating conditions is seen to be a difficult parameter to control in view of the limited options for effective cooling. It is therefore essential to take serious note of this aspect, to arrest rapid deterioration of bushing. The degradation of dry-type insulation such as RIP is often due to thermal stress. The long time performance thereof, depends strongly, on the maximum operating temperature. With this in view, the Authors have developed a theoretical model and computational method to study the temperature distribution in the body of insulation. The Authors consider that the basis for the model as being the temperature and electric stress aided AC conductivity. The ensuing heat balance (continuity) equations in 2-D cylindrical geometry are treated as a Dirichelet-Neumann boundary value problem.

A simple model for spatial disaggregation of evaporative fraction: Comparative study with thermal sharpened land surface temperature data over India

[1] Evaporative fraction (EF) is a measure of the amount of available energy at the earth surface that is partitioned into latent heat flux. The currently operational thermal sensors like the Moderate Resolution Imaging Spectroradiometer (MODIS) on satellite platforms provide data only at 1000 m, which constraints the spatial resolution of EF estimates. A simple model (disaggregation of evaporative fraction (DEFrac)) based on the observed relationship between EF and the normalized difference vegetation index is proposed to spatially disaggregate EF. The DEFrac model was tested with EF estimated from the triangle method using 113 clear sky data sets from the MODIS sensor aboard Terra and Aqua satellites. Validation was done using the data at four micrometeorological tower sites across varied agro-climatic zones possessing different land cover conditions in India using Bowen ratio energy balance method. The root-mean-square error (RMSE) of EF estimated at 1000 m resolution using the triangle method was 0.09 for all the four sites put together. The RMSE of DEFrac disaggregated EF was 0.09 for 250 m resolution. Two models of input disaggregation were also tried with thermal data sharpened using two thermal sharpening models DisTrad and TsHARP. The RMSE of disaggregated EF was 0.14 for both the input disaggregation models for 250 m resolution. Moreover, spatial analysis of disaggregation was performed using Landsat-7 (Enhanced Thematic Mapper) ETM+ data over four grids in India for contrasted seasons. It was observed that the DEFrac model performed better than the input disaggregation models under cropped conditions while they were marginally similar under non-cropped conditions.

An extended finite-element model coupled with level set method for analysis of growth of corrosion pits in metallic structures

Mass balance between metal and electrolytic solution, separated by a moving interface, in stable pit growth results in a set of governing equations which are solved for concentration field and interface position (pit boundary evolution). The interface experiences a jump discontinuity in metal concentration. The extended finite-element model (XFEM) handles this jump discontinuity by using discontinuous-derivative enrichment formulation, eliminating the requirement of using front conforming mesh and re-meshing after each time step as in the conventional finite-element method. However, prior interface location is required so as to solve the governing equations for concentration field for which a numerical technique, the level set method, is used for tracking the interface explicitly and updating it over time. The level set method is chosen as it is independent of shape and location of the interface. Thus, a combined XFEM and level set method is developed in this paper. Numerical analysis for pitting corrosion of stainless steel 304 is presented. The above proposed model is validated by comparing the numerical results with experimental results, exact solutions and some other approximate solutions. An empirical model for pitting potential is also derived based on the finite-element results. Studies show that pitting profile depends on factors such as ion concentration, solution pH and temperature to a large extent. Studying the individual and combined effects of these factors on pitting potential is worth knowing, as pitting potential directly influences corrosion rate.

A model for the classification and survey of clock synchronization protocols in WSNs

Clock synchronization in wireless sensor networks (WSNs) assures that sensor nodes have the same reference clock time. This is necessary not only for various WSN applications but also for many system level protocols for WSNs such as MAC protocols, and protocols for sleep scheduling of sensor nodes. Clock value of a node at a particular instant of time depends on its initial value and the frequency of the crystal oscillator used in the sensor node. The frequency of the crystal oscillator varies from node to node, and may also change over time depending upon many factors like temperature, humidity, etc. As a result, clock values of different sensor nodes diverge from each other and also from the real time clock, and hence, there is a requirement for clock synchronization in WSNs. Consequently, many clock synchronization protocols for WSNs have been proposed in the recent past. These protocols differ from each other considerably, and so, there is a need to understand them using a common platform. Towards this goal, this survey paper categorizes the features of clock synchronization protocols for WSNs into three types, viz, structural features, technical features, and global objective features. Each of these categories has different options to further segregate the features for better understanding. The features of clock synchronization protocols that have been used in this survey include all the features which have been used in existing surveys as well as new features such as how the clock value is propagated, when the clock value is propagated, and when the physical clock is updated, which are required for better understanding of the clock synchronization protocols in WSNs in a systematic way. This paper also gives a brief description of a few basic clock synchronization protocols for WSNs, and shows how these protocols fit into the above classification criteria. In addition, the recent clock synchronization protocols for WSNs, which are based on the above basic clock synchronization protocols, are also given alongside the corresponding basic clock synchronization protocols. Indeed, the proposed model for characterizing the clock synchronization protocols in WSNs can be used not only for analyzing the existing protocols but also for designing new clock synchronization protocols. (C) 2014 Elsevier B.V. All rights reserved.

Conjugate Model for Heat and Mass Transfer of Porous Wall in the High Temperature Gas Flow

Heat and mass transfer of a porous permeable wall in a high temperature gas dynamical flow is considered. Numerical simulation is conducted on the ground of the conjugate mathematical model which includes filtration and heat transfer equations in a porous body and boundary layer equations on its surface. Such an approach enables one to take into account complex interaction between heat and mass transfer in the gasdynamical flow and in the structure subjected to this flow. The main attention is given to the impact of the intraporous heat transfer intensity on the transpiration cooling efficiency.

Analysis of Kic and its temperature dependence of metals by a simplified dislocation model

The relative Kic values of metals are calculated with a simplified dislocation model. It is found that the ratio of KIIc to KIc and the temperature dependence of fracture toughness of some metals estimated with this model are consistent with the experimental results.