Circular dichroism and nuclear magnetic resonance studies on the complexation of valinomycin with calcium

Complexation of valinomycin (VM) with the divalent cation Ca2+ in a lipophilic solvent, acetonitrile (CH3-CN), has been studied by using circular dichroism and proton and carbon- 13 nuclear magnetic resonance (‘H NMR and I3C NMR). From analyses of the spectral data, it is concluded that VM forms a 2:l (peptideion-peptide) sandwich complex with Ca2+, at low concentration of VM. At moderate conocentrations of the salt, in addition to the sandwich complex, an equimolar (1:l) complex different from those observed for potassium and sodium is also observed. At very large concentrations of the calcium salt, the data suggested a complex with a conformation similar to that of the free VM in polar solvents. Possible conformations for the sandwich and the equimolar VM-calcium complexes are proposed.

Hydrogen- 1 and Carbon- 13 Magnetic Resonance Studies of Nonactin-Calcium Complex

For an understanding of the cation selectivity and general binding characteristics of macrotetralide antibiotic nonactin (NA) with ions of different sizes and charges, the nature of binding of divalent cation, Ca2+, to NA and conformation of the NA-Ca2+ complex have been studied by use of 270-MHz proton nuclear magnetic resonance ('H NMR) and carbon-13 nuclear magnetic resonance (13C NMR). The calcium ion induced significantly large changes in chemical shifts for H7, H2, H3, and H5 protons of NA and relatively small changes for H18 and H2' protons. Changes in I3C chemical shift were quite large for carbonyl carbon, C,; it is noteworthy that in the NA-K+ complex, H2 and H2' protons practically do not show any change during complexation and carbonyl carbon shows a much smaller chemical shift change.

1H and 13C nuclear magnetic resonance studies on X-537A (lasalocid-A)-calcium complexes: observation of a sandwich complex

Studies on the conformational and binding characteristics of the ionophoric antibiotic X-537A (lasalocid-A)�calcium ion complexes have been carried out in deuteriated acetonitrile (CD3 CN) using proton and carbon-13 nuclear magnetic resonance (1 H and 13C n.m.r.) spectroscopy. Detailed analysis of the salt-induced chemical shifts at various X-537A to calcium concentration ratios indicated that X-537A forms charged complexes with calcium with 2 : 1 and 1 : 1 stoicheiometries. The conformational model for the complex based on the n.m.r. data showed that the calcium ion is preferentially bound to one end of the molecule, which is binding to three oxygen atoms, the other end (the salicylic acid part) being relatively free. In the 2 : 1 (sandwich) complex, the calcium ion is sandwiched between two X-537A molecules with three oxygen atoms binding to it from each molecule.

Possible involvement of a calcium-stimulated ATP-hydrolyzing activity associated with mycobacteriophage I3 in the DNA injection process.

Ca2+ ions are necessary for the successful propagation of mycobacteriophage I3. An assay for the phage DNA release in the presence of an isolated cell wall preparation from the host was established, and in this system Ca2+ ions also stimulated the release of DNA. The inhibition of phage DNA injection caused by Tween 80 (polyoxyethylene sorbitan monooleate), a nonionic detergent routinely used in mycobacterial cultures, was reversed by Ca2+. The presence of a phage-associated ATP-hydrolyzing activity was demonstrated. This enzyme was stimulated by Ca2+ ions and inhibited by Tween 80. From this and the behavior of the two agents at the level of DNA injection, as well as the fact that phage I3 has a contractile tail structure, we conclude that the phage-associated ATPase is involved in the DNA injection process.

Thermal decomposition of doped calcium hydroxide for chemical energy storage

Thermal decomposition of Ca(OH)2 with and without additives has been experimentally investigated for its application as a thermochemical energy storage system. The homogeneous reaction model gives a satisfactory fit for the kinetic data on pure and Ni(OH)2---, Zn(OH)2--- and Al(OH)3---doped Ca(OH)2 and the order of reaction is 0.76 in all cases except for the Al(OH)3-doped sample for which the decomposition is zero order. These additives are shown not only to enhance the reaction rate but also to reduce the decomposition temperature significantly. Some models for solid decomposition reactions, and possible mechanisms in the decomposition of solids containing additives, are also discussed.

Requirement for calcium ions in mycobacteriophage I3 DNA injection and propagation

Ca2+ ions are absolutely necessary for the propagation of mycobacteriophage I3 in synthetic medium. These ions are required for successful infection of the host and during the entire span of the intracellular development of the phage. A direct assay of the phage DNA injection using 32[P] labelled phage, showns that Ca2+ ions are necessary for the injection process. The injection itself is a slow process and takes 15 min to complete at 37°C. The bacteria infected in presence of Ca2+ tend to abort if the ions are subsequently withdrawn from the growth medium. The effect of calcium withdrawal is maximally felt during the early part of the latent period; however, later supplementation of Ca2+ ions salvage phage production and the mature phage progeny appear after a delayed interval, proportional to the time of addition of Ca2+.

Thermodynamics of metal ion binding and denaturation of a calcium binding protein from Entamoeba histolytica

The thermodynamics of tie binding of calcium and magnesium ions to a calcium binding protein from Entamoeba histolytica was investigated by isothermal titration calorimetry (ITC) in 20 mM MOPS buffer (pH 7.0) at 20 degrees C. Enthalpy titration curves of calcium show the presence of four Ca2+ binding sites, There exist two low-affinity sites for Ca2+, both of which are exothermic in nature and with positive cooperative interaction between them. Two other high affinity sites for Ca2+ exist of which one is endothermic and the other exothermic, again with positive cooperative interaction. The binding constants for Ca2+ at the four sites have been verified by a competitive binding assay, where CaBP competes with a chromophoric chelator 5, 5'-Br-2 BAPTA to bind Ca2+ and a Ca2+ titration employing intrinsic tyrosine fluorescence of the protein, The enthalpy of titration of magnesium in the absence of calcium is single site and endothermic in nature. In the case of the titrations performed using protein presaturated with magnesium, the amount of heat produced is altered. Further, the interaction between the high-affinity sites changes to negative cooperativity. No exchange of heat was observed throughout the addition of magnesium in the presence of 1 mM calcium, Titrations performed on a cleaved peptide comprising the N-terminus and the central linker show the existence of two Ca2+ specific sites, These results indicate that this CaBP has one high-affinity Ca-Mg site, one high-affinity Ca-specific site, and two low-affinity Ca-specific sites. The thermodynamic parameters of the binding of these metal ions were used to elucidate the energetics at the individual site(s) and the interactions involved therein at various concentrations of the denaturant, guanidine hydrochloride, ranging from 0.05 to 6.5 M. Unfolding of the protein was also monitored by titration calorimetry as a function of the concentration of the denaturant. These data show that at a GdnHCl concentration of 0.25 M the binding affinity for the Mg2+ ion is lost and there are only two sites which can bind to Ca2+, with substantial loss cooperativity. At concentrations beyond 2.5 M GdnHCl, at which the unfolding of the tertiary structure of this protein is observed by near UV CD spectroscopy, the binding of Ca2+ ions is lost. We thus show that the domain containing the two low-affinity sites is the first to unfold in the presence of GdnHCl. Control experiments with change in ionic strength by addition of KCI in the range 0.25-1 M show the existence of four sites with altered ion binding parameters.

Spatial variation of sequestered calcium in the multicellular stage of Dictyostelium discoideum as assayed by chlortetracycline fluorescence

Abstract. We have used chlortetracycline (CTC) as a fluorescent probe to detect the distribution of sequestered calcium in multicellular stages of Dictyostelium discoideum. Tips of late aggregates, slugs and early culminating masses fluoresce very strongly. Most of the fluorescence is intracellular in origin and emanates from a small number of intense punctate sources. The sources correspond in part to autophagic vacuoles viz. neutral-red staining, acidic digestive vesicles, and may also include intracellular organelles; cytoplasmic fluorescence is much weaker in comparison. The level of fluorescence drops in the middle portion of slugs and rises again in the posteriormost region, though not to as high a level as in the tip. This holds good irrespective of whether CTC is applied only in the neighbourhood of the aggregate centre, only in the aggregate periphery, or to the whole aggregate. We infer that there must be a good deal of mixing in the stages leading from aggregation to slug formation; thus the serial order in which cells enter an aggregate does not bear any relation to their ultimate fates. The other implication of our study is that calcium sequestration is much more extensive in prestalk and anterior-like cells than in prespore cells. These findings are discussed with regard to possible implications for pattern formation.

Glycodelin A triggers T cell apoptosis through a novel calcium-independent galactose-binding lectin activity

Glycodelin A (GdA) is one of the progesterone inducible endometrial factors that protect the fetal semiallograft from maternal immune rejection. The immumoregulatory effects of GdA are varied, with diverse effects on the fate and function of most immune cell types. Its effects on T cells are particularly relevant as it is capable of regulating T cell activation, differentiation, as well as apoptosis. We have previously reported that GdA triggers mitochondrial stress and apoptosis in activated T cells by a mechanism that is distinct and independent of its effects on T cell activation. In this study we describe the characterization of a cell surface receptor for GdA on T cells. Our results reveal a novel calcium-independent galactose-binding lectin activity of GdA, which is responsible for its apoptogenic function. This discovery adds GdA to a select group of soluble immunoregulatory lectins that operate within the feto-placental compartment, the only other members being the galectin family proteins. We also report for the first time that both CD4(+) and CD8(+) T cell subsets are equally susceptible to inhibition with GdA, mediated by its novel lectin activity. We demonstrate that GdA selectively recognizes complex-type N-linked glycans on T cell surface glycoproteins. and propose that the galectin-1 glycoprotein receptor CD7 maybe a novel target for GdA on T cells. This study, for the first time, links the lectin activity of GdA to its biological function.

Pseudo-time marching schemes for inverse problems in structural health assessment and medical imaging

We propose a self-regularized pseudo-time marching strategy for ill-posed, nonlinear inverse problems involving recovery of system parameters given partial and noisy measurements of system response. While various regularized Newton methods are popularly employed to solve these problems, resulting solutions are known to sensitively depend upon the noise intensity in the data and on regularization parameters, an optimal choice for which remains a tricky issue. Through limited numerical experiments on a couple of parameter re-construction problems, one involving the identification of a truss bridge and the other related to imaging soft-tissue organs for early detection of cancer, we demonstrate the superior features of the pseudo-time marching schemes.

High Resolution Surface Imaging Using a Carbon Nanotube Array with Pointed Height distribution

In this paper we discuss a new technique to image the surfaces of metallic substrates using field emission from a pointed array of carbon nanotubes (CNTs). We consider a pointed height distribution of the CNT array under a diode configuration with two side gates maintained at a negative potential to obtain a highly intense beam of electrons localized at the center of the array. The CNT array on a metallic substrate is considered as the cathode and the test substrate as the anode. Scanning the test Substrate with the cathode reveals that the field emission current is highly sensitive to the surface features with nanometer resolution. Surface features of semi-circular, triangular and rectangular geometries (projections and grooves) are considered for simulation. This surface scanning/mapping technique can be applied for surface roughness measurements with nanoscale accuracy. micro/nano damage detection, high precision displacement sensors, vibrometers and accelerometers. among other applications.

Evaluation of Moderate-Resolution Imaging Spectroradiometer (MODIS) collection 004 (C004) aerosol retrievals at Kanpur, Indo-Gangetic basin

[1] We have compared the spectral aerosol optical depth (AOD, tau lambda) and aerosol fine mode fraction (AFMF) of Collection 004 (C004) derived from Moderate-Resolution Imaging Spectroradiometer (MODIS) on board National Aeronautics and Space Administration's (NASA) Terra and Aqua platforms with that obtained from Aerosol Robotic Network (AERONET) at Kanpur (26.45 degrees N, 80.35 degrees E), India for the period 2001-2005. The spatially-averaged (0.5 degrees x 0.5 degrees centered at AERONET sunphotometer) MODIS Level-2 aerosol parameters (10 km at nadir) were compared with the temporally averaged AERONET-measured AOD (within +/- 30 minutes of MODIS overpass). We found that MODIS systematically overestimated AOD during the pre-monsoon season (March to June, known to be influenced by dust aerosols). The errors in AOD at 0.66 mu m were correlated with the apparent reflectance at 2.1 mu m (rho*(2.1)) which MODIS C004 uses to estimate the surface reflectance in the visible channels (rho(0.47) = rho*(2.1)/ 4, rho(0.66) = rho*(2.1)/ 2). The large errors in AOD (Delta tau(0.66) > 0.3) are found to be associated with the higher values of rho*(2.1) (0.18 to 0.25), where the uncertainty in the ratios of reflectance is large (Delta rho(0.66) +/- 0.04, Delta rho(0.47) +/- 0.02). This could have resulted in lower surface reflectance, higher aerosol path radiance and thus lead to overestimation in AOD. While MODIS-derived AFMF has binary distribution (1 or 0) with too low (AFMF < 0.2) during dust-loading period, and similar to 1 for the rest of the retrievals, AERONET showed range of values (0.4 to 0.9). The errors in tau(0.66) were also high in the scattering angle range 110 degrees - 140 degrees, where the optical effects of nonspherical dust particles are different from that of spherical particles.

Improved k-t BLAST for fast fMR imaging

A popular dynamic imaging technique, k-t BLAST (ktB) is studied here for BAR imaging. ktB utilizes correlations in k-space and time, to reconstruct the image time series with only a fraction of the data. The algorithm works by unwrapping the aliased Fourier conjugate space of k-t (y-f-space). The unwrapping process utilizes the estimate of the true y-f-space, by acquiring densely sampled low k-space data. The drawbacks of this method include separate training scan, blurred training estimates and aliased phase maps. The proposed changes are incorporation of phase information from the training map and using generalized-series-extrapolated training map. The proposed technique is compared with ktB on real fMRI data. The proposed changes allow for ktB to operate at an acceleration factor of 6. Performance is evaluated by comparing activation maps obtained using reconstructed images. An improvement of up to 10 dB is observed in thePSNR of activation maps. Besides, a 10% reduction in RMSE is obtained over the entire time series of fMRI images. Peak improvement of the proposed method over ktB is 35%, averaged over five data sets. (C)2010 Elsevier Inc. All rights reserved.

Polarisation studies on the Raman spectra of cubic crystals - Part II: Calcium fluoride and potassium aluminium sulphate

Polarisation characters of the Raman lines of calcium fluoride (fluorspar) and potassium aluminium sulphate (alum) were investigated under the following conditions. Unpolarised light was incident normally on a face of the crystal making an angle 22.5° with a cubic face and the light scattered transversely along a cubic axis was analysed by a double image prism kept with its principal axes inclined at 45° to the vertical. Under these conditions the depolarisation factors of the Raman lines belonging to the totally symmetric (A), the doubly degenerate (E) and the triply degenerate (F) modes should be respectively =1, >1 and <1. The characteristic Raman line of CaF2 at 322 cm-1 exhibited a depolarisation value less than 1, showing thereby that the corresponding mode is a triply degenerate one (F). The Raman lines observed in the spectrum of K-alum were also classified and the results were compared with those given by previous investigators using standard crystal orientations.