840 research outputs found
Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)
Introduction:
In this study, quasi-three-dimensional (3D) microwell patterns were fabricated with poly (l-lactic acid) for the development of cell-based assays, targeting voltage-gated calcium channels (VGCCs).
Methods and materials:
SH-SY5Y human neuroblastoma cells were interfaced with the microwell patterns and found to grow as two dimensional (2D), 3D, and near two dimensional (N2D), categorized on the basis of the cells’ location in the pattern. The capability of the microwell patterns to support 3D cell growth was evaluated in terms of the percentage of the cells in each growth category. Cell spreading was analyzed in terms of projection areas under light microscopy. SH-SY5Y cells’ VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium GreenTM-1. The expression of L-type calcium channels was evaluated using immunofluorescence staining with DM-BODIPY.
Results:
It was found that cells within the microwells, either N2D or 3D, showed more rounded shapes and less projection areas than 2D cells on flat poly (l-lactic acid) substrates. Also, cells in microwells showed a significantly lower VGCC responsiveness than cells on flat substrates, in terms of both response magnitudes and percentages of responsive cells, upon depolarization with 50 mM K+. This lower VGCC responsiveness could not be explained by the difference in L-type calcium channel expression. For the two patterns addressed in this study, N2D cells consistently exhibited an intermediate value of either projection areas or VGCC responsiveness between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness.
Conclusion:
These results suggest that the pattern structure and therefore the cell growth characteristics were critical factors in determining cell VGCC responsiveness and thus provide an approach for engineering cell functionality in cell-based assay systems and tissue engineering scaffolds
ADAM15 mediates upregulation of Claudin-1 expression in breast cancer cells
A Disintegrin and Metalloproteinase-15 (ADAM15) is a transmembrane protein involved in protein ectodomain shedding, cell adhesion and signalling. We previously cloned and characterised alternatively spliced variants of ADAM15 that differ in their intracellular domains and demonstrated correlation of the expression of specific variants with breast cancer prognosis. In this study we have created isogenic cell panels (MDA-MB-231 and MCF-7) expressing five ADAM15 variants including wildtype and catalytically inactive forms. The expression of ADAM15 isoforms in MDA-MB-231 cells led to cell clustering to varying degree, without changes in EMT markers vimentin, slug and E-cadherin. Analysis of tight junction molecules revealed ADAM15 isoform specific, catalytic function dependent upregulation of Claudin-1. The expression of ADAM15A, and to a lesser degree of C and E isoforms led to an increase in Claudin-1 expression in MDA-MB-231 cells, while ADAM15B had no effect. In MCF-7 cells, ADAM15E was the principal variant inducing Claudin-1 expression. Sh-RNA mediated down-regulation of ADAM15 in ADAM15 over-expressing cells reduced Claudin-1 levels. Additionally, downregulation of endogenous ADAM15 expression in T47D cells by shRNA reduced endogenous Claudin-1 expression confirming a role for ADAM15 in regulating Claudin-1 expression. The PI3K/Akt/mTOR pathway was involved in regulating Claudin-1 expression downstream of ADAM15. Immunofluorescence analysis of MDA-MB-231 ADAM15A expressing cells showed Claudin-1 at cell-cell junctions, in the cytoplasm and nuclei. ADAM15 co-localised with Claudin-1 and ZO1 at cell-cell junctions. Immunoprecipitation analysis demonstrated complex formation between ADAM15 and ZO1/ZO2. These findings highlight the importance of ADAM15 Intra Cellular Domain-mediated interactions in regulating substrate selection and breast cancer cell phenotype
Single Spin Asymmetry in Polarized Proton-Proton Elastic Scattering at GeV
We report a high precision measurement of the transverse single spin
asymmetry at the center of mass energy GeV in elastic
proton-proton scattering by the STAR experiment at RHIC. The was measured
in the four-momentum transfer squared range \GeVcSq, the region of a significant interference between the
electromagnetic and hadronic scattering amplitudes. The measured values of
and its -dependence are consistent with a vanishing hadronic spin-flip
amplitude, thus providing strong constraints on the ratio of the single
spin-flip to the non-flip amplitudes. Since the hadronic amplitude is dominated
by the Pomeron amplitude at this , we conclude that this measurement
addresses the question about the presence of a hadronic spin flip due to the
Pomeron exchange in polarized proton-proton elastic scattering.Comment: 12 pages, 6 figure
Longitudinal double-spin asymmetry and cross section for inclusive neutral pion production at midrapidity in polarized proton collisions at sqrt(s) = 200 GeV
We report a measurement of the longitudinal double-spin asymmetry A_LL and
the differential cross section for inclusive Pi0 production at midrapidity in
polarized proton collisions at sqrt(s) = 200 GeV. The cross section was
measured over a transverse momentum range of 1 < p_T < 17 GeV/c and found to be
in good agreement with a next-to-leading order perturbative QCD calculation.
The longitudinal double-spin asymmetry was measured in the range of 3.7 < p_T <
11 GeV/c and excludes a maximal positive gluon polarization in the proton. The
mean transverse momentum fraction of Pi0's in their parent jets was found to be
around 0.7 for electromagnetically triggered events.Comment: 6 pages, 3 figures, submitted to Phys. Rev. D (RC
High non-photonic electron production in + collisions at = 200 GeV
We present the measurement of non-photonic electron production at high
transverse momentum ( 2.5 GeV/) in + collisions at
= 200 GeV using data recorded during 2005 and 2008 by the STAR
experiment at the Relativistic Heavy Ion Collider (RHIC). The measured
cross-sections from the two runs are consistent with each other despite a large
difference in photonic background levels due to different detector
configurations. We compare the measured non-photonic electron cross-sections
with previously published RHIC data and pQCD calculations. Using the relative
contributions of B and D mesons to non-photonic electrons, we determine the
integrated cross sections of electrons () at 3 GeV/10 GeV/ from bottom and charm meson decays to be = 4.0({\rm
stat.})({\rm syst.}) nb and =
6.2({\rm stat.})({\rm syst.}) nb, respectively.Comment: 17 pages, 17 figure
Evolution of the differential transverse momentum correlation function with centrality in Au+Au collisions at GeV
We present first measurements of the evolution of the differential transverse
momentum correlation function, {\it C}, with collision centrality in Au+Au
interactions at GeV. {\it C} exhibits a strong dependence
on collision centrality that is qualitatively similar to that of number
correlations previously reported. We use the observed longitudinal broadening
of the near-side peak of {\it C} with increasing centrality to estimate the
ratio of the shear viscosity to entropy density, , of the matter formed
in central Au+Au interactions. We obtain an upper limit estimate of
that suggests that the produced medium has a small viscosity per unit entropy.Comment: 7 pages, 4 figures, STAR paper published in Phys. Lett.
Elliptic flow of electrons from heavy-flavor hadron decays in Au+Au collisions at 200, 62.4, and 39 GeV
We present measurements of elliptic flow () of electrons from the decays
of heavy-flavor hadrons () by the STAR experiment. For Au+Au collisions
at 200 GeV we report , for transverse momentum
() between 0.2 and 7 GeV/c using three methods: the event plane method
({EP}), two-particle correlations ({2}), and four-particle
correlations ({4}). For Au+Au collisions at = 62.4 and
39 GeV we report {2} for GeV/c. {2} and {4} are
non-zero at low and intermediate at 200 GeV, and {2} is consistent
with zero at low at other energies. The {2} at the two lower beam
energies is systematically lower than at 200 GeV for
GeV/c. This difference may suggest that charm quarks interact less
strongly with the surrounding nuclear matter at those two lower energies
compared to GeV.Comment: Version accepted by PR
Measurements of Dihadron Correlations Relative to the Event Plane in Au+Au Collisions at GeV
Dihadron azimuthal correlations containing a high transverse momentum (\pt)
trigger particle are sensitive to the properties of the nuclear medium created
at RHIC through the strong interactions occurring between the traversing parton
and the medium, i.e. jet-quenching. Previous measurements revealed a strong
modification to dihadron azimuthal correlations in Au+Au collisions with
respect to \pp\ and \dAu\ collisions. The modification increases with the
collision centrality, suggesting a path-length dependence to the jet-quenching
effect. This paper reports STAR measurements of dihadron azimuthal correlations
in mid-central (20-60\%) Au+Au collisions at \snn=200~GeV as a function of
the trigger particle's azimuthal angle relative to the event plane,
\phis=|\phit-\psiEP|. The azimuthal correlation is studied as a function of
both the trigger and associated particle \pt. The subtractions of the
combinatorial background and anisotropic flow, assuming Zero Yield At Minimum
(\zyam), are described. The away-side correlation is strongly modified, and the
modification varies with \phis, which is expected to be related to the
path-length that the away-side parton traverses. The pseudo-rapidity (\deta)
dependence of the near-side correlation, sensitive to long range \deta
correlations (the ridge), is also investigated. The ridge and jet-like
components of the near-side correlation are studied as a function of \phis.
The ridge appears to drop with increasing \phis while the jet-like component
remains approximately constant. ...Comment: 50 pages, 39 figures, 6 table
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