Abstract:The aim of this study was to prepare a novel targeting drug delivery system for 2-Methoxyestradiol (2ME) in order to improve the clinical application of this antitumor drug. It is based in nanoparticles (NPs) of titanium dioxide (TiO 2 ) coated with polyethylene glycol (PEG) and loaded with 2ME. A complete IR and Raman characterization have been made to confirm the formation of TiO 2 -PEG-2ME composite. Vibrational modes have been assigned for TiO 2 , PEG, and 2ME and functionalized TiO 2 -PEG and TiO 2 -PEG-2ME. The observed variation in peak position of FTIR and Raman of each for these composites has been elucidated in terms of intermolecular interactions between PEG-2ME and TiO 2 , obtaining step-by-step the modification processes that were attributed to the conjugation of PEG and 2ME to TiO 2 NPs. Modifying TiO 2 NPs with PEG loaded with the 2ME drug revealed that the titanium dioxide nanocarrier possesses an effective adsorption capability, and we discuss their potential application as a system of drug delivery.

A method to determine the three components (3C) of the velocity field in a micro volume (3D) using a single camera is proposed. The technique is based on tracking the motion of individual particles to exclude errors due to depth of correlation (DOC) and spatial averaging as in μPIV (micro particle image velocimetry). The depth position of the particles is coded by optical distortions initiated by a cylindrical lens in the optical setup. To estimate the particle positions, a processing algorithm was developed based on continuous wavelet analysis and autocorrelation. This algorithm works robustly and gives accurate results comparable to multi-camera systems (tomographic PIV, V3V). Particle tracking was applied to determine the full 3C velocity vector in the volume without the error due to spatial averaging and DOC, which are inherent limitations in μPIV due to the interrogation windows size and volume illumination. To prove the applicability, measurements were performed in a straight channel with a cross section of 500 × 500 μm 2 . The depth of the measurement volume in the viewing direction was chosen to be 90 μm in order to resolve the near-wall gradients. The three-dimensional velocity distribution of the whole channel could be resolved clearly by using wave front deformation particle tracking velocimetry.

Astigmatism particle tracking velocimetry (APTV) is a method to determine three components (3C) of the velocity field in a volume (3D) using a single camera. The depth position of the particles is coded by optical distortions caused by a cylindrical lens in the optical setup. This technique is particularly suited for microfluidic applications as measurement errors due to spatial averaging and depth of correlation, typically encountered with μPIV approaches, are eliminated so that the measurement precision is enhanced. Unfortunately, the current state of the technique is limited by the small measurement region achievable with the current calibration procedures as well as by higher order image aberrations (Cierpka et al 2010 Meas. Sci. Technol. 21 045401). In order to extend the size of the measurement volume and to account for all image aberrations, a new intrinsic calibration procedure, based on the imaging function of the particles, is proposed in the paper at hand. It provides an extended measurement depth, taking into account all image aberrations. In this work, the calibration procedure was applied to a μPIV arrangement but could also be implemented on macroscopic experimental setups. The calibration procedure is qualified with synthetic data as well as Poiseuille flow in a straight rectangular micro-channel with a cross-sectional area of 200 ×  500 µm2. The three-dimensional velocity distribution of the whole channel was resolved via APTV with uncertainties of 0.9% and 3.7% of the centerline velocity, uc, for the in-plane and out-of-plane components, respectively. Further investigations using different cylindrical-lens focal lengths, magnifications and particle sizes provide information about achievable measurement depths and help to design and adapt the optimal system for the desired experiment.

Glioblastoma multiforme (GBM) cells are characterised by their extreme chemoresistance. The activity of multiple-drug resistance (MDR) transporters that extrude antitumor drugs from cells plays the most important role in this phenomenon. To date, the mechanism controlling the expression and activity of MDR transporters is poorly understood. Activity of the enzyme ecto-5 0 -nucleotidase (CD73) in tumor cells, which hydrolyses AMP to adenosine, has been linked to immunosuppression and prometastatic effects in breast cancer and to the proliferation of glioma cells. In this study, we identify a high expression of CD73 in surgically resected samples of human GBM. In primary cultures of GBM, inhibition of CD73 activity or knocking down its expression by siRNA reversed the MDR phenotype and cell viability was decreased up to 60% on exposure to the antitumoral drug vincristine. This GBM chemosensitization was caused by a decrease in the expression and activity of the multiple drug associated protein 1 (Mrp1), the most important transporter conferring multiple drug resistance in these cells. Using pharmacological modulators, we have recognized the adenosine A 3 receptor subtype in mediation of the chemoresistant phenotype in these cells. In conclusion, we have determined that the activity of CD73 to trigger adenosine signaling sustains chemoresistant phenotype in GBM cells. ORIGINAL RESEARCH ARTICLE 602 J o u r n a l o f J o u r n a l o f Cellular Physiology Cellular Physiology ß 2 0 1 2 W I L E Y P E R I O D I C A L S , I N C .

The depth of correlation (DOC) is an experimental parameter, introduced to quantify the thickness of the measurement volume and thus the depth resolution in microscopic particle image velocimetry (lPIV). The theory developed to estimate the value of the DOC relies on some approximations that are not always verified in actual experiments, such as a single thin-lens optical system. In many practical lPIV experiments, a deviation of the actual DOC from its nominal value can be expected, due for instance to additional components present in the optical path of the microscope or to the use of image preprocessing before the PIV evaluation. In the presented paper, the effect of real particle image intensity distribution and image preprocessing on the thickness of the measurement volume is investigated. This is performed studying the defocusing of tracer particles and the DOC-related bias error present in lPIV measurements in a Poiseuille flow. The analysis shows that the DOC predicted using the conventional formulas can be significantly smaller than its actual value. To overcome this problem, the use of an effective NA determined experimentally from the curvature of the image autocorrelations is proposed. The accuracy of this approach to properly predict the actual size of DOC is discussed and validated on the experimental data. The effectiveness of image preprocessing to reduce the DOC-related bias error is tested and discussed as well.

Glioblastoma multiforme (GBM) is a brain tumour characterised by a remarkably high chemoresistance and infiltrating capability. To date, chemotherapy with temozolomide has contributed only poorly to improved survival rates in patients. One of the most important mechanisms of chemoresistance comes about through the activity of certain proteins from the ATP-binding cassette superfamily that extrudes antitumour drugs, or their metabolites, from cells. We identify an increased expression of the multiple drug resistance-associated protein 1 (Mrp1) in glioblastoma multiforme biopsies and in T98G and G44 cell lines. The activity of this transporter was also confirmed by measuring the extrusion of the fluorescent substrate CFDA. The sensitivity of GBM cells was low upon exposure to temozolomide, vincristine and etoposide, with decreases in cell viability of below 20% seen at therapeutic concentrations of these drugs. However, combined exposure to vincristine or etoposide with an inhibitor of Mrp1 efficiently decreased cell viability by up to 80%. We conclude that chemosensitization of cells with inhibitors of Mrp1 activity might be an efficient tool for the treatment of human GBM.

A diffusion-based microreactor system operated with a reaction volume of 8 μL is presented and characterized to intensify the process understanding in microscale cultivations. Its potential as screening tool for biological processes is evaluated. The advantage of the designed microbioreactor is the use for the continuous cultivation mode by integrating online measurement technique for dissolved oxygen (DO) and optical density (OD). A further advantage is the broaden application for biological systems. The bioreactor geometry was chosen to achieve homogeneous flow during continuous process operation. The device consisted of a microstructured top layer made of poly(dimethylsiloxane) (PDMS), which was designed and fabricated using UV-depth and soft lithography assembled with a glass bottom. CFD simulation data used for geometry design were verified via microparticle-image-velocimetry (μPIV). In the used microreactor geometry no concentration gradients occurred along the entire reaction volume because of rapid diffusive mixing, the homogeneous medium flow inside the growth chamber of the microreactor could be realized. Undesirable bubble formation before and during operation was reduced by using degassed medium as well as moistened and moderate incident air flow above the gas permeable PDMS membrane. Because of this a passive oxygen supply of the culture medium in the device is ensured by diffusion through the PDMS membrane. The oxygen supply itself was monitored online via integrated DO sensors based on a fluorescent dye complex. An adequate overall volumetric oxygen transfer coefficient K(L)a as well as mechanical stability of the device were accomplished for a membrane thickness of 300 μm. Experimental investigations considering measurements of OD (online) and several metabolite concentrations (offline) in a modified Verduyn medium. The used model organism Saccharomyces cerevisiae DSM 2155 tended to strong reactor wall growth resembling a biofilm.

A combination of cutting edge developments is presented to characterize three-dimensional (3D) temperature and velocity fields in microscopic flows. An emulsion of non-encapsulated thermo-liquid crystal (TLC) micro spheres, with a narrow size distribution is used to track the flow's motion and temperature distribution. A state-of-the-art light engine, which combines the spectrum of six light pipes, provides a balanced illumination which allows for strong and detectable color patterns across the TLC's temperature response range. Lastly, the ability of the TLC material to reflect select wavelength bands with an unchanging and independent circular polarization chirality is exploited by a filter that blocks background noise, while exclusively transmitting the color signal of the TLC particles. This approach takes advantage of the peculiar physical properties of TLCs to allow the estimation of individual TLC particle's 3D position, for the first time, using Astigmatism Particle Tracking Velocimetry (APTV).