A simplified protocol for detecting two systemic bait markers (Rhodamine B and iophenoxic acid) in small mammals

We developed a method of quantifying levels of fluorescence in the whiskers of wild stoats (Mustela erminea) using fluorescence microscopy and Axiovision 3.0.6.1 software. The method allows for discrimination between natural fluorescence present in or on a whisker, and the fluorescence resulting from the ingestion of the systemic marker Rhodamine B (RB), although some visual judgement is still required. We also developed a new high performance liquid chromatography (HPLC) protocol for detecting the systemic marker iophenoxic acid (IPA) in the blood of laboratory rats (Rattus norvegicus) and wild stoats. With this method, the blood of an animal that has consumed IPA can be tested for the presence of the foreign IPA compound itself. This is a more reliable test than the previous method, which measured the raised level of natural blood protein-bound iodine correlated with IPA absorption. The quantity of blood required from animal subjects is very small (10 μl), so the testing is less intrusive and the method can be extended to smaller species. The extraction technique uses methanol, rather than acids and heavy metal salts, thereby simplifying the procedure. Recovery of IPA is quantitative, giving a highly reliable reading. In experiments on captive rats the IPA method proved successful. Of 12 positively marked carcasses, two that had not been frozen for the 24 h before blood samples were taken showed relatively lower IPA levels. The same IPA detection method, as well as the whisker analysis for RB, was applied successfully to a population of wild stoats to which both Rhodamine B and IPA were made available at bait stations. The presence of both bait markers was detectable in rats for at least 21 days and in stoats for at least 27 days

Geometric approach to Fletcher's ideal penalty function

Original article can be found at: www.springerlink.com Copyright Springer. [Originally produced as UH Technical Report 280, 1993]In this note, we derive a geometric formulation of an ideal penalty function for equality constrained problems. This differentiable penalty function requires no parameter estimation or adjustment, has numerical conditioning similar to that of the target function from which it is constructed, and also has the desirable property that the strict second-order constrained minima of the target function are precisely those strict second-order unconstrained minima of the penalty function which satisfy the constraints. Such a penalty function can be used to establish termination properties for algorithms which avoid ill-conditioned steps. Numerical values for the penalty function and its derivatives can be calculated efficiently using automatic differentiation techniques.Peer reviewe

De novo design of a reversible phosphorylation-dependent switch for membrane targeting

Modules that switch protein-protein interactions on and off are essential to develop synthetic biology; for example, to construct orthogonal signaling pathways, to control artificial protein structures dynamically, and for protein localization in cells or protocells. In nature, the E. coli MinCDE system couples nucleotide-dependent switching of MinD dimerization to membrane targeting to trigger spatiotemporal pattern formation. Here we present a de novo peptide-based molecular switch that toggles reversibly between monomer and dimer in response to phosphorylation and dephosphorylation. In combination with other modules, we construct fusion proteins that couple switching to lipid-membrane targeting by: (i) tethering a 'cargo' molecule reversibly to a permanent membrane 'anchor'; and (ii) creating a 'membrane-avidity switch' that mimics the MinD system but operates by reversible phosphorylation. These minimal, de novo molecular switches have potential applications for introducing dynamic processes into designed and engineered proteins to augment functions in living cells and add functionality to protocells. The ability to dynamically control protein-protein interactions and localization of proteins is critical in synthetic biological systems. Here the authors develop a peptide-based molecular switch that regulates dimer formation and lipid membrane targeting via reversible phosphorylation.The authors thank the Biochemistry Core Facility of the Max Planck Institute of Biochemistry for LC-MS and CD spectroscopy services, Stefan Pettera and Stephan Uebel for assistance with peptide synthesis and analytical HPLC, and Katharina Nakel for assistance with cloning

Detection of brain functional-connectivity difference in post-stroke patients using group-level covariance modeling

Functional brain connectivity, as revealed through distant correlations in the signals measured by functional Magnetic Resonance Imaging (fMRI), is a promising source of biomarkers of brain pathologies. However, establishing and using diagnostic markers requires probabilistic inter-subject comparisons. Principled comparison of functional-connectivity structures is still a challenging issue. We give a new matrix-variate probabilistic model suitable for inter-subject comparison of functional connectivity matrices on the manifold of Symmetric Positive Definite (SPD) matrices. We show that this model leads to a new algorithm for principled comparison of connectivity coefficients between pairs of regions. We apply this model to comparing separately post-stroke patients to a group of healthy controls. We find neurologically-relevant connection differences and show that our model is more sensitive that the standard procedure. To the best of our knowledge, these results are the first report of functional connectivity differences between a single-patient and a group and thus establish an important step toward using functional connectivity as a diagnostic tool

Investigating the Magnetospheres of Rapidly Rotating B-type Stars

Recent spectropolarimetric surveys of bright, hot stars have found that ~10% of OB-type stars contain strong (mostly dipolar) surface magnetic fields (~kG). The prominent paradigm describing the interaction between the stellar winds and the surface magnetic field is the magnetically confined wind shock (MCWS) model. In this model, the stellar wind plasma is forced to move along the closed field loops of the magnetic field, colliding at the magnetic equator, and creating a shock. As the shocked material cools radiatively it will emit X-rays. Therefore, X-ray spectroscopy is a key tool in detecting and characterizing the hot wind material confined by the magnetic fields of these stars. Some B-type stars are found to have very short rotational periods. The effects of the rapid rotation on the X-ray production within the magnetosphere have yet to be explored in detail. The added centrifugal force due to rapid rotation is predicted to cause faster wind outflows along the field lines, leading to higher shock temperatures and harder X-rays. However, this is not observed in all rapidly rotating magnetic B-type stars. In order to address this from a theoretical point of view, we use the X-ray Analytical Dynamical Magnetosphere (XADM) model, originally developed for slow rotators, with an implementation of new rapid rotational physics. Using X-ray spectroscopy from ESA's XMM-Newton space telescope, we observed 5 rapidly rotating B-type stars to add to the previous list of observations. Comparing the observed X-ray luminosity and hardness ratio to that predicted by the XADM allows us to determine the role the added centrifugal force plays in the magnetospheric X-ray emission of these stars.Comment: IAUS Conference Proceeding

de Haas-van Alphen effect investigation of the electronic structure of Al substituted MgB_2

We report a de Haas-van Alphen (dHvA) study of the electronic structure of Al doped crystals of MgB$_2$. We have measured crystals with $\sim 7.5$% Al which have a $T_c$ of 33.6 K, ($\sim 14$% lower than pure MgB$_2$). dHvA frequencies for the $\sigma$ tube orbits in the doped samples are lower than in pure MgB$_2$, implying a $16\pm2$ reduction in the number of holes in this sheet of Fermi surface. The mass of the quasiparticles on the larger $\sigma$ orbit is lighter than the pure case indicating a reduction in electron-phonon coupling constant $\lambda$. These observations are compared with band structure calculations, and found to be in excellent agreement.Comment: 4 pages with figure

Shubnikov-de Haas oscillations in YBa_2Cu_4O_8

We report the observation of Shubnikov-de Haas oscillations in the underdoped cuprate superconductor YBa$_2$Cu$_4$O$_8$ (Y124). For field aligned along the c-axis, the frequency of the oscillations is $660\pm 30$ T, which corresponds to $\sim 2.4$ % of the total area of the first Brillouin zone. The effective mass of the quasiparticles on this orbit is measured to be $2.7\pm0.3$ times the free electron mass. Both the frequency and mass are comparable to those recently observed for ortho-II YBa$_2$Cu$_3$O$_{6.5}$ (Y123-II). We show that although small Fermi surface pockets may be expected from band structure calculations in Y123-II, no such pockets are predicted for Y124. Our results therefore imply that these small pockets are a generic feature of the copper oxide plane in underdoped cuprates.Comment: v2: Version of paper accepted for publication in Physical Review Letters. Only minor changes to the text and reference

Transverse Magnetoresistance of GaAs/AlGaAs Heterojunctions in the Presence of Parallel Magnetic Fields

We have calculated the resistivity of a GaAs\slash AlGaAs heterojunction in the presence of both an in--plane magnetic field and a weak perpendicular component using a semiclassical Boltzmann transport theory. These calculations take into account fully the distortion of the Fermi contour which is induced by the parallel magnetic field. The scattering of electrons is assumed to be due to remote ionized impurities. A positive magnetoresistance is found as a function of the perpendicular component, in good qualitative agreement with experimental observations. The main source of this effect is the strong variation of the electronic scattering rate around the Fermi contour which is associated with the variation in the mean distance of the electronic states from the remote impurities. The magnitude of the positive magnetoresistance is strongly correlated with the residual acceptor impurity density in the GaAs layer. The carrier lifetime anisotropy also leads to an observable anisotropy in the resistivity with respect to the angle between the current and the direction of the in--plane magnetic field.Comment: uuencoded file containing a 26 page RevTex file and 14 postscript figures. Submitted to Phys. Rev.