3,495 research outputs found
Stochastic Ratchet Mechanisms for Replacement of Proteins Bound to DNA
Experiments indicate that unbinding rates of proteins from DNA can depend on
the concentration of proteins in nearby solution. Here we present a theory of
multi-step replacement of DNA-bound proteins by solution-phase proteins. For
four different kinetic scenarios we calculate the depen- dence of protein
unbinding and replacement rates on solution protein concentration. We find (1)
strong effects of progressive 'rezipping' of the solution-phase protein onto
DNA sites liberated by 'unzipping' of the originally bound protein; (2) that a
model in which solution-phase proteins bind non-specifically to DNA can
describe experiments on exchanges between the non specific DNA- binding
proteins Fis-Fis and Fis-HU; (3) that a binding specific model describes
experiments on the exchange of CueR proteins on specific binding sites.Comment: \`a paraitre en PHys. Rev. Lett. june 201
Thermodynamics of beta-amyloid fibril formation
Amyloid fibers are aggregates of proteins. They are built out of a peptide
called --amyloid (A) containing between 41 and 43 residues,
produced by the action of an enzyme which cleaves a much larger protein known
as the Amyloid Precursor Protein (APP). X-ray diffraction experiments have
shown that these fibrils are rich in --structures, whereas the shape of
the peptide displays an --helix structure within the APP in its
biologically active conformation. A realistic model of fibril formation is
developed based on the seventeen residues A12--28 amyloid peptide, which
has been shown to form fibrils structurally similar to those of the whole
A peptide. With the help of physical arguments and in keeping with
experimental findings, the A12--28 monomer is assumed to be in four
possible states (i.e., native helix conformation, --hairpin, globular
low--energy state and unfolded state). Making use of these monomeric states,
oligomers (dimers, tertramers and octamers) were constructed. With the help of
short, detailed Molecular Dynamics (MD) calculations of the three monomers and
of a variety of oligomers, energies for these structures were obtained. Making
use of these results within the framework of a simple yet realistic model to
describe the entropic terms associated with the variety of amyloid
conformations, a phase diagram can be calculated of the whole many--body
system, leading to a thermodynamical picture in overall agreement with the
experimental findings. In particular, the existence of micellar metastable
states seem to be a key issue to determine the thermodynamical properties of
the system
Understanding the determinants of stability and folding of small globular proteins from their energetics
The results of minimal model calculations suggest that the stability and the
kinetic accessibility of the native state of small globular proteins are
controlled by few "hot" sites. By mean of molecular dynamics simulations around
the native conformation, which simulate the protein and the surrounding solvent
at full--atom level, we generate an energetic map of the equilibrium state of
the protein and simplify it with an Eigenvalue decomposition. The components of
the Eigenvector associated with the lowest Eigenvalue indicate which are the
"hot" sites responsible for the stability and for the fast folding of the
protein. Comparison of these predictions with the results of mutatgenesis
experiments, performed for five small proteins, provide an excellent agreement
Fine frequency shift of sigle vortex entrance and exit in superconducting loops
The heat capacity of an array of independent aluminum rings has been
measured under an external magnetic field using highly sensitive
ac-calorimetry based on a silicon membrane sensor. Each superconducting vortex
entrance induces a phase transition and a heat capacity jump and hence
oscillates with . This oscillatory and non-stationary behaviour
measured versus the magnetic field has been studied using the Wigner-Ville
distribution (a time-frequency representation). It is found that the
periodicity of the heat capacity oscillations varies significantly with the
magnetic field; the evolution of the period also depends on the sweeping
direction of the field. This can be attributed to a different behavior between
expulsion and penetration of vortices into the rings. A variation of more than
15% of the periodicity of the heat capacity jumps is observed as the magnetic
field is varied. A description of this phenomenon is given using an analytical
solution of the Ginzburg-Landau equations of superconductivity
Model adaptation enriched with an anisotropic mesh spacing for nonlinear equations: application to environmental and CFD problems
Goal of this paper is to suitably combine a model with an anisotropic mesh
adaptation for the numerical simulation of nonlinear advection-diffusion-reaction systems and incompressible flows in ecological and environmental applications. Using the reduced-basis method terminology, the proposed approach leads to a noticeable computational saving of the online phase with respect to the resolution of the reference model on nonadapted grids. The search of a suitable adapted model/mesh pair is to be meant, instead, in an offline fashion
Tadalafil modulates aromatase activity and androgen receptor expression in a human osteoblastic cell in vitro model
Purpose Phosphodiesterase type-5 inhibitor (PDE5i)
tadalafil administration in men with erectile dysfunction is
associated with increased testosterone/estradiol ratio, leading
to hypothesize a potential increased effect of androgen
action on target tissues. We aimed to characterize, in a cellular
model system in vitro, the potential modulation of
aromatase and sex steroid hormone receptors upon exposure
to tadalafil (TAD).
Methods Human osteoblast-like cells SAOS-2 were chosen
as an in vitro model system since osteoblasts are target
of steroid hormones. Cells were tested for viability upon
TAD exposure, which increased cell proliferation. Then,
cells were treated with/without TAD for several times to
evaluate potential modulation in PDE5, aromatase (ARO),
androgen (AR) and estrogen (ER) receptor expression.
Results Osteoblasts express significant levels of both PDE5
mRNA and protein. Exposure of cells to increasing concentrations
of TAD (10−8–10−7 M) decreased PDE5 mRNA and
protein expression. Also, TAD inhibited ARO mRNA and
protein expression leading to an increase in testosterone levels
in the supernatants. Interestingly, TAD increased total AR mRNA and protein expression and decreased ERα, with an
increased ratio of AR/ER, suggesting preferential androgenic
vs estrogenic pathway activation.
Conclusions Our results demonstrate for the first time
that TAD decreases ARO expression and increases AR
protein expression in human SAOS-2, strongly suggesting
a new control of steroid hormones pathway by PDE5i.
These findings might represent the first evidence of translational
actions of PDE5i on AR, which leads to hypothesize
a growing relevance of this molecule in men with
prostate cancer long-term treated with TAD for sexual
rehabilitation.
Keywords Tadalafil · Osteoblasts · Aromatase · Androgen
receptor · Estrogen recepto
A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior
Background: One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells.
Methods: High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50–100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated.
Results: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells.
Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay
Development and reproduction of Cataclysta lemnata, a potential natural enemy of the invasive alien duckweed Lemna minuta in Italy
Life cycle of the aquatic moth Cataclysta lemnata (Lepidoptera: Crambidae) was studied in laboratory conditions to obtain a basic biological knowledge useful for predicting the possible success of the herbivorous larvae of this insect as potential control agents in limiting the spread of the invasive American duckweed Lemna minuta (Alismatales: Araceae) in Italy. The multivoltinism of C. lemnata, as well as the high overall emergence from the pupal stage (85%), the high success in mating among the formed couples (>90%), and the high number of larvae born from each egg laying (on average 310 individuals), suggest that the insect can be successfully bred in the laboratory for the purposes of an augmentative biological control. Under experimental conditions, larvae developed in 23 days (through six larval instars, distinguishable by cephalic capsule dimensions) and pupae in 10, with no difference in duration between females and males. The larval phase resulted longer than the adult one (23 vs 10 days), therefore it can be considered the most suitable stage for releasing the insect in field for biocontrol purposes. Indeed, the larvae having a herbivorous diet might consume large amount of the invasive plant, contrarily to the adult phase which is focused exclusively on reproduction. The results emerged not only allow to contribute to the knowledge on aquatic lepidoptera that are scarcely known, but also support the effectiveness of a possible protocol for an augmentative biological control of the invasive alien duckweed L. minuta
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