The activation of fibroblast growth factors (FGFs) by glycosaminoglycans: influence of the sulfation pattern on the biological activity of FGF-1

Six synthetic heparin-like oligosaccharides have been used to investigate the effect of the oligosaccharide sulfation pattern on the stimulation of acidic fibroblast growth factor (FGF-1) induced mitogenesis signaling and the biological significance of FGF-1 trans dimerization in the FGF-1 activation process. It has been found that some molecules with a sulfation pattern that does not contain the internal trisaccharide motif, which has been proposed for high affinity for FGF-1, stimulate FGF-1 more efficiently than those with the structure of the regular region of heparin. In contrast to regular region oligosaccharides, in which the sulfate groups are distributed on both sides of their helical three-dimensional structures, the molecules containing this particular sulfation pattern display the sulfate groups only on one side of the helix. These results and the fact that these oligosaccharides do not promote FGF-1 dimerization according to sedimentation-equilibrium analysis, confirm the importance of negative-charge distribution in the activation process and strongly suggest that FGF dimerization is not a general and absolute requirement for biological activity.     

NMR and modeling studies of protein-carbohydrate interactions: synthesis, three-dimensional structure, and recognition properties of a minimum hevein domain with binding affinity for chitooligosaccharides

HEV32, a 32-residue, truncated hevein lacking eleven C-terminal amino acids, was synthesized by solid-phase methodology and correctly folded with three cysteine bridge pairs. The affinities of HEV32 for small chitin fragments--in the forms of N,N',N"-triacetylchitotriose ((GlcNAc)3) (millimolar) and N,N',N",N"',N",N"'-hexaacetylchitohexaose ((GlcNAc)6) (micromolar)--as measured by NMR and fluorescence methods, are comparable with those of native hevein. The HEV32 ligand-binding process is enthalpy driven, while entropy opposes binding. The NMR structure of ligand-bound HEV32 in aqueous solution was determined to be highly similar to the NMR structure of ligand-bound hevein. Solvated molecular-dynamics simulations were performed in order to monitor the changes in side-chain conformation of the binding site of HEV32 and hevein upon interaction with ligands. The calculations suggest that the Trp21 side-chain orientation of HEV32 in the free form differs from that in the bound state; this agrees with fluorescence and thermodynamic data. HEV32 provides a simple molecular model for studying protein-carbohydrate interactions and for understanding the physiological relevance of small native hevein domains lacking C-terminal residues.     

Characterization of Candida albicans orthologue of the Saccharomyces cerevisiae signal-peptidase-subunit encoding gene SPC3

The Candida albicans orthologue of the SPC3 gene, which encodes one of the subunits essential for the activity of the signal peptidase complex in Saccharomyces cerevisiae, was isolated by complementation of a thermosensitive mutation in the S. cerevisiae SEC61 gene. The cloned gene (CaSPC3) encodes a putative protein of 192 amino acids that contains one potential membrane-spanning region and shares significant homology with the corresponding products from mammalian (Spc22/23p) and yeast (Spc3p) cells. CaSPC3 is essential for cell viability, since a hemizygous strain containing a single copy of CaSPC3 under control of the methionine-repressible MET3 promoter did not grow in the presence of methionine and cysteine. The cloned gene could rescue the phenotype associated with a spc3 mutation in S. cerevisiae, indicating that it is the true C. albicans orthologue of SPC3. However, in contrast with results previously described for its S. cerevisiae orthologue, CaSPC3 was not able to complement the thermosensitive growth associated with a mutation in the SEC11 gene. The heterologous complementation of the sec61 mutant suggests that Spc3p could play a role in the interaction that it is known to occur between the translocon (Sec61 complex) and the signal peptidase complex, at the endoplasmic reticulum membrane.     

Shelf life extension of beef retail cuts subjected to an advanced vacuum skin packaging system

Vacuum skin packaging (VSP) is a new and promising technology that includes the instantaneous heating of the upper packaging film immediately before its descent over the meat surface. In the present paper we evaluated this technology for the packaging of a wide variety of beef retail cuts and compared this with traditional vacuum packaging (TVP) performed in parallel. Shelf-life and quality evolution was estimated by analysing three spoilage bacterial groups, an ubiquitous psychrotrophic food pathogen, and three biochemical and physico-chemical attributes throughout 40 days of storage at 4 °C. Statistically-significant lower microbial counts were obtained for all microbial parameters and meat types. The VSP lot exhibited lower pH. This, however, involved a lower water holding capacity and a higher degree of firmness, the latter indicating a limited tenderisation process in the VSP batch. Extended aging times in the carcasses were studied to improving the tenderness of VSP samples. The extension of the aging process prior to VSP may be useful for improving the tenderness of certain meat types and to improve the sensory quality of meat cuts processed under this system.     

Fatty acid profile of traditional soymilk

Soymilk is a water extract of soybeans, closely resembling dairy milk in physical appearance and composition. Most fatty acids in soybean and its derivates are unsaturated, and therefore susceptible to oxidation. The aim of this study was to evaluate the possible effect of the thermal conditions during the elaboration process of soymilk on its fatty acid profile. For this, the fatty acid composition of soymilk, okara (soymilk residue), and soybean were studied by gas chromatography with flame ionization detection (GC-FID). No major differences in the fatty acid patterns were found.     

Simulation of electron energy loss near-edge structure at the Al and N K edges and Al L(23) edge in cubic aluminium nitride

Calculations of electron energy near edge structures (ELNES) are compared with experimental data obtained in a high-resolution transmission electron microscope. This study concerns small precipitates of aluminium nitride in low carbon steel. The ELNES technique allows to clearly establish that these precipitates crystallize in a cubic rather than in a hexagonal crystallographic cell. The influence on simulated spectra of different parameters are investigated: the size of the atomic shell and its relation with the electron inelastic mean free path. We also examine the influence of the core hole and the sensitivity to cell parameters. We particularly examine the Al L(23) near edge structure and features relating to the different transition channels (A(1g), E(g) and T(2g)). Results of a multiple scattering and band structure calculations using ICXANES and WIEN97 codes, respectively, are compared in the region from 0 to 30 eV above the edge onsets. Both calculations are in a rather good agreement.     

Thermal AFM: a thermopile case study

In this work, an atomic force microscope (AFM) with an integrated thermal sensor has been used to obtain the local spatial distribution of temperatures in a micromachined thermopile with submicron resolution. In this communication, we will show how the dimensional, structural and functional characteristics of a thermopile suits well with the requirements for AFM thermal imaging, and how a deeper insight of the thermopile operation can be gained with the aid of these advanced scanning probe-based tools.     

An analytic method to predict the thermal map of cryosurgery iceballs in MR images

This paper presents a newly developed method to estimate, in magnetic resonance (MR) images, the temperatures reached within the volume of an iceball produced by a cryogenic probe. Building on the direct measurements of the MR signal intensity and its correlation with independent temperature variations at the phase transition from liquid to solid, the thermal information embedded in the images was accessed. The volume and diameter of the growing iceball were estimated from a time series of MR images. Using regressions over the volume in the time and thermal domains, this method predicted the cryogenic temperatures beyond the range of sensitivity of the MR signal itself. We present a validation of this method in samples of gelatin and ex vivo pig liver. Temperature predictions are shown to agree with independent thermosensor readings over a range extending from 20 degrees C down to -65 degrees C, with an average error of less than 6 degrees C.     

Denoising functional MR images: a comparison of wavelet denoising and Gaussian smoothing

We present a general wavelet-based denoising scheme for functional magnetic resonance imaging (fMRI) data and compare it to Gaussian smoothing, the traditional denoising method used in fMRI analysis. One-dimensional WaveLab thresholding routines were adapted to two-dimensional (2-D) images, and applied to 2-D wavelet coefficients. To test the effect of these methods on the signal-to-noise ratio (SNR), we compared the SNR of 2-D fMRI images before and after denoising, using both Gaussian smoothing and wavelet-based methods. We simulated a fMRI series with a time signal in an active spot, and tested the methods on noisy copies of it. The denoising methods were evaluated in two ways: by the average temporal SNR inside the original activated spot, and by the shape of the spot detected by thresholding the temporal SNR maps. Denoising methods that introduce much smoothness are better suited for low SNRs, but for images of reasonable quality they are not preferable, because they introduce heavy deformations. Wavelet-based denoising methods that introduce less smoothing preserve the sharpness of the images and retain the original shapes of active regions. We also performed statistical parametric mapping on the denoised simulated time series, as well as on a real fMRI data set. False discovery rate control was used to correct for multiple comparisons. The results show that the methods that produce smooth images introduce more false positives. The less smoothing wavelet-based methods, although generating more false negatives, produce a smaller total number of errors than Gaussian smoothing or wavelet-based methods with a large smoothing effect.