Modeling the growth of multicellular cancer spheroids in a\ud bioengineered 3D microenvironment and their treatment with an\ud anti-cancer drug

A critical step in the dissemination of ovarian cancer cells is the formation of multicellular spheroids from cells shed from the primary tumor. The objectives of this study were to establish and validate bioengineered three-dimensional (3D) microenvironments for culturing ovarian cancer cells in vitro and simultaneously to develop computational models describing the growth of multicellular spheroids in these bioengineered matrices. Cancer cells derived from human epithelial ovarian carcinoma were embedded within biomimetic hydrogels of varying stiffness and cultured for up to 4 weeks. Immunohistochemistry was used to quantify the dependence of cell proliferation and apoptosis on matrix stiffness, long-term culture and treatment with the anti-cancer drug paclitaxel.\ud \ud Two computational models were developed. In the first model, each spheroid was treated as an incompressible porous medium, whereas in the second model the concept of morphoelasticity was used to incorporate details about internal stresses and strains. Each model was formulated as a free boundary problem. Functional forms for cell proliferation and apoptosis motivated by the experimental work were applied and the predictions of both models compared with the output from the experiments. Both models simulated how the growth of cancer spheroids was influenced by mechanical and biochemical stimuli including matrix stiffness, culture time and treatment with paclitaxel. Our mathematical models provide new perspectives on previous experimental results and have informed the design of new 3D studies of multicellular cancer spheroids

Growth of confined cancer spheroids: a combined experimental and mathematical modelling approach

We have integrated a bioengineered three-dimensional platform by generating multicellular cancer spheroids in a controlled microenvironment with a mathematical model to investigate\ud confined tumour growth and to model its impact on cellular processes

High speed video capture for mobile phone cameras

We consider an electromechanical model for the operation of a voice coil motor in a mobile phone camera, with the aim of optimizing how a lens can be moved to a desired focusing motion. Although a methodology is developed for optimizing lens shift, there is some concern about the experimentally-determined model parameters that are at our disposal. Central to the model is the value of the estimated magnetic force constant, Kf: its value determines how far it is actually possible to move lens, but it appears that, from the value given, it would not be possible to shift the lens through the displacements desired. Furthermore, earlier experiments have also estimated the value of the back EMF constant, Kg , to be roughly five times greater than Kf, even though we present two theoretical arguments that show that Kf = Kg: a conclusion supported by readily-available manufacturers’ data

Implicit reconstructions of thin leaf surfaces from large, noisy point clouds

Thin surfaces, such as the leaves of a plant, pose a significant challenge for implicit surface reconstruction techniques, which typically assume a closed, orientable surface. We show that by approximately interpolating a point cloud of the surface (augmented with off-surface points) and restricting the evaluation of the interpolant to a tight domain around the point cloud, we need only require an orientable surface for the reconstruction. We use polyharmonic smoothing splines to fit approximate interpolants to noisy data, and a partition of unity method with an octree-like strategy for choosing subdomains. This method enables us to interpolate an N-point dataset in O(N) operations. We present results for point clouds of capsicum and tomato plants, scanned with a handheld device. An important outcome of the work is that sufficiently smooth leaf surfaces are generated that are amenable for droplet spreading simulations

The cytoplasmic carbonic anhydrases βCA2 and βCA4 are required for optimal plant growth at low CO\u3csub\u3e2\u3c/sub\u3e

© 2016 American Society of Plant Biologists. All Rights Reserved. Carbonic anhydrases (CAs) are zinc metalloenzymes that interconvert CO2 and HCO3-. In plants, both α-and β-type CAs are present. We hypothesize that cytoplasmic bCAs are required to modulate inorganic carbon forms needed in leaf cells for carbonrequiring reactions such as photosynthesis and amino acid biosynthesis. In this report, we present evidence that βCA2 and βCA4 are the two most abundant cytoplasmic CAs in Arabidopsis (Arabidopsis thaliana) leaves. Previously, βCA4 was reported to be localized to the plasma membrane, but here, we show that two forms of βCA4 are expressed in a tissue-specific manner and that the two proteins encoded by βCA4 localize to two different regions of the cell. Comparing transfer DNA knockout lines with wild-type plants, there was no reduction in the growth rates of the single mutants, βca2 and βca4. However, the growth rate of the double mutant, βca2βca4, was reduced significantly when grown at 200μL L-1 CO2. The reduction in growth of the double mutant was not linked to a reduction in photosynthetic rate. The amino acid content of leaves from the double mutant showed marked reduction in aspartate when compared with the wild type and the single mutants. This suggests the cytoplasmic CAs play an important but not previously appreciated role in amino acid biosynthesis

The carbon concentrating mechanism in Chlamydomonas reinhardtii: Finding the missing pieces

The photosynthetic, unicellular green alga, Chlamydomonas reinhardtii, lives in environments that often contain low concentrations of CO2 and HCO3-, the utilizable forms of inorganic carbon (Ci). C. reinhardtii possesses a carbon concentrating mechanism (CCM) which can provide suitable amounts of Ci for growth and development. This CCM is induced when the CO2 concentration is at air levels or lower and is comprised of a set of proteins that allow the efficient uptake of Ci into the cell as well as its directed transport to the site where Rubisco fixes CO2 into biomolecules. While several components of the CCM have been identified in recent years, the picture is still far from complete. To further improve our knowledge of the CCM, we undertook a mutagenesis project where an antibiotic resistance cassette was randomly inserted into the C. reinhardtii genome resulting in the generation of 22,000 mutants. The mutant collection was screened using both a published PCR-based approach (Gonzalez-Ballester et al. 2011) and a phenotypic growth screen. The PCR-based screen did not rely on a colony having an altered growth phenotype and was used to identify colonies with disruptions in genes previously identified as being associated with the CCM-related gene. Eleven independent insertional mutations were identified in eight different genes showing the usefulness of this approach in generating mutations in CCM-related genes of interest as well as identifying new CCM components. Further improvements of this method are also discussed. © 2014 Springer Science+Business Media Dordrecht

Second-Order Coherence Across the Brillouin Lasing Threshold

Brillouin-Mandelstam scattering is one of the most accessible nonlinear optical phenomena and has been widely studied since its theoretical discovery one hundred years ago. The scattering mechanism is a three-wave mixing process between two optical fields and one acoustic field and has found a broad range of applications spanning microscopy to ultra-narrow-linewidth lasers. Building on the success of utilizing this nonlinearity at a classical level, a rich avenue is now being opened to explore Brillouin scattering within the paradigm of quantum optics. Here, we take a key step in this direction by employing quantum optical techniques yet to be utilized for Brillouin scattering to characterize the second-order coherence of Stokes scattering across the Brillouin lasing threshold. We use a silica microsphere resonator and single-photon counters to observe the expected transition from bunched statistics of thermal light below the lasing threshold to Poissonian statistics of coherent light above the threshold. Notably, at powers approaching the lasing threshold, we also observe super-thermal statistics, which arise due to instability and a ``flickering'' in and out of lasing as the pump field is transiently depleted. The statistics observed across the transition, including the ``flickering'', are a result of the full nonlinear three-wave mixing process and cannot be captured by a linearized model. These measurements are in good agreement with numerical solutions of the three-wave Langevin equations and are well demarcated by analytical expressions for the instability and the lasing thresholds. These results demonstrate that applying second-order-coherence and photon-counting measurements to Brillouin scattering provides new methods to advance our understanding of Brillouin scattering itself and progress toward quantum-state preparation and characterization of acoustic modes.Comment: Main (8 pages, 2 figures) + Supplementary (6 pages, 1 figures), Submitte