1,094 research outputs found
Surface flow profiles for dry and wet granular materials by Particle Tracking Velocimetry; the effect of wall roughness
Two-dimensional Particle Tracking Velocimetry (PTV) is a promising technique
to study the behaviour of granular flows. The aim is to experimentally
determine the free surface width and position of the shear band from the
velocity profile to validate simulations in a split-bottom shear cell geometry.
The position and velocities of scattered tracer particles are tracked as they
move with the bulk flow by analyzing images. We then use a new technique to
extract the continuum velocity field, applying coarse-graining with the
postprocessing toolbox MercuryCG on the discrete experimental PTV data. For
intermediate filling heights, the dependence of the shear (or angular) velocity
on the radial coordinate at the free surface is well fitted by an error
function. From the error function, we get the width and the centre position of
the shear band. We investigate the dependence of these shear band properties on
filling height and rotation frequencies of the shear cell for dry glass beads
for rough and smooth wall surfaces. For rough surfaces, the data agrees with
the existing experimental results and theoretical scaling predictions. For
smooth surfaces, particle-wall slippage is significant and the data deviates
from the predictions. We further study the effect of cohesion on the shear band
properties by using small amount of silicon oil and glycerol as interstitial
liquids with the glass beads. While silicon oil does not lead to big changes,
glycerol changes the shear band properties considerably. The shear band gets
wider and is situated further inward with increasing liquid saturation, due to
the correspondingly increasing trend of particles to stick together
Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry.
Multidrug resistance (MDR) in tumour cells is often caused by the overexpression of the plasma membrane drug transporter P-glycoprotein (P-gp) or the recently discovered multidrug resistance-associated protein (MRP). In this study we investigated the specificity and sensitivity of the fluorescent probes rhodamine 123 (R123), daunorubicin (DNR) and calcein acetoxymethyl ester (calcein-AM) in order to detect the function of the drug transporters P-gp and MRP, using flow cytometry. The effects of modulators on the accumulation and retention of these probes were compared in several pairs of sensitive and P-gp- as well as MRP-overexpressing cell lines. R123, in combination with the modulator PSC833, provided the most sensitive test for detecting P-gp-mediated resistance. Moreover, in a 60 min drug accumulation assay R123 can be regarded as a P-gp-specific probe, since R123 is not very efficiently effluxed by MRP. In contrast to R123, a 60 min DNR or calcein-AM accumulation test could be used to detect MRP-mediated resistance. The MRP-specific modulator genistein could be used in combination with DNR, but not with calcein-AM. Vincristine (VCR) can be used to increase the cellular uptake of calcein-AM in MDR cells, but is not specific for MRP. Thus, although the combination of DNR with genistein appeared to be as sensitive as the combination of calcein-AM with VCR, the former may be used to probe specific MRP activity whereas the latter provides a combined (P-gp + MRP) functional MDR parameter. With these functional assays the role and relative importance of P-gp and MRP can be studied in, for example, haematological malignancies
Characterization of a clonal human colon adenocarcinoma line intrinsically resistant to doxorubicin.
Intrinsic low-level resistance to anti-cancer drugs is a major problem in the treatment of gastrointestinal malignancies. To address the problem presented by intrinsically resistant tumours, we have isolated two monoclonal lines from LoVo human colon adenocarcinoma cells: LoVo/C7, which is intrinsically resistant to doxorubicin (DOX); and LoVo/C5, which shows the same resistance index for DOX as the mixed parental cell population. For comparison, we have included in the study a LoVo-resistant line selected by continuous exposure to DOX and expressing a typical multidrug resistant (MDR) phenotype. In these cell lines we have studied the expression and/or activity of a number of proteins, including P-glycoprotein 170 (P-gp), multidrug resistance-associated protein (MRP), lung resistance-related protein (LRP), glutathione (GSH)-dependent enzymes and protein kinase C (PKC) isoforms, which have been implicated in anti-cancer drug resistance. Intracellular DOX distribution has been assessed by confocal microscopy. The results of the present study indicate that resistance in LoVo/C7 cells cannot be attributed to alterations in P-gp, LRP or GSH/GSH-dependent enzyme levels. Increased expression of MRP, accompanied by alterations in the subcellular distribution of DOX, has been observed in LoVo/C7 cells; changes in PKC isoform pattern have been detected in both intrinsically and pharmacologically resistant cells
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