Short-term changes in carbon isotope composition of soluble carbohydrates and starch: from canopy leaves to the root system

Changes in the 13C discrimination of current leaf photosynthesis might have profound impacts on root respiratory substrates. Therefore, the aim of this study was (1) to refine a method for the isolation of root and leaf starch and soluble sugars (neutral fraction) for stable carbon isotope analysis and (2) to assess the short-term temporal variability of the C isotope composition (delta13C) of starch and of the neutral fraction of beech roots and leaves at different canopy heights. An existing method for isolating starch for stable C isotope analysis based on enzymatic hydrolysis was modified to account for the low starch content of the samples. This was achieved by removing the enzyme (alpha-amylase) by ultrafiltration after the hydrolysis, resulting in very low carbon blanks. The neutral fraction was separated from organic acids and cations by ion-exchange chromatography. An anion-exchange resin in the [HCO3]--form was chosen that ensured high precision of C blanks. Beech leaves at 5, 10 and 20 m above the forest floor as well as roots were sampled six times during a day/night cycle in July 2003. Delta13C values of bulk material, starch and the neutral fraction increased from the lower to the higher canopy with mean differences between 5 and 20 m of 3.8, 3.4 and 2.7 per thousand for the delta13C values of starch, neutral fraction and bulk foliage, respectively. The delta13C value of foliar starch increased from the morning to the afternoon and decreased during the night, but diurnal differences (up to 3.1 per thousand) were only statistically significant for leaves sampled at 5 and 10 m height. In roots, no diurnal variation in the delta13C of starch was observed during the short time frame of one day and the delta13C of the neutral fraction did not differ between samples taken at 16:30 and 22:00. Calculated delta13C values of starch, which was mobilised during the night, were more positive than the total starch (all sampling times pooled) in leaves. Furthermore, the delta13C values of mobilised starch were approximately 5 per thousand more positive than that of the mobilised neutral fraction. Hence, the delta13C of potential sources for export from canopy leaves to roots varied considerably in their C isotope composition.     

Preparation and characterization of thin films of SiO(x) on gold substrates for surface plasmon resonance studies

We report here on the fabrication and characterization of stable thin films of amorphous silica (SiO(x)) deposited on glass slides coated with a 5 nm adhesion layer of titanium and 50 nm of gold, using the plasma-enhanced chemical vapor deposition (PECVD) technique. The resulting surfaces were characterized using atomic force microscopy (AFM), ellipsometry, contact angle measurements, and surface plasmon resonance (SPR). AFM analysis indicates that homogeneous films of silica with low roughness were formed on the gold surface. The deposited silica films showed excellent stability in different solvents and in piranha solution. There was no significant variation in the thickness or in the SPR signal after these harsh treatments. The Au/SiO(x) interfaces were investigated for their potential applications as new surface plasmon resonance sensor chips. Silica films with thicknesses up to 40 nm allowed visualization of the surface plasmon effect, while thicker films resulted in the loss of the SPR characteristics. SPR allowed further the determination of the silica thickness and was compared to ellipsometric results. Chemical treatment of the SiO(x) film with piranha solution led to the generation of silanol surface groups that have been coupled with a trichlorosilane.     

Nanoscale zinc antimonides: synthesis and phase stability

Highly crystalline single-phase nanoparticles of the important thermoelectric materials Zn4Sb3 and ZnSb were prepared from solvochemically activated powders of elemental zinc and elemental antimony. Low-temperature reactions with reaction temperatures of 275-300 degrees C were applied using an excess of elemental zinc. The nanoscale thermoelectrics obtained were characterized by X-ray powder diffraction, transmission electron microscopy, and thermal analysis. nc-Zn4Sb3 showed particle sizes of 50-70 nm, whereas particle sizes of 15-20 nm were observed for nc-ZnSb. Calorimetric investigations showed an increased heat capacity, Cp, for nc-Zn4Sb3 with respect to the bulk material which could be reduced to the bulk value by annealing nc-Zn4Sb3 at 190 degrees C. Interestingly, nc-Zn4Sb3 showed exothermic decomposition into zinc-poorer ZnSn at 196 degrees C in an open system, indicating that Zn4Sb3 is metastable in nanocrystalline form at room temperature.     

Monomeric iron(II) hydroxo and iron(III) dihydroxo complexes stabilized by intermolecular hydrogen bonding

Using the multidentate ligand bis(N-methylimidazol-2-yl)-3-methylthiopropanol (L), the mononuclear iron(II) hydroxo and iron(III) dihydroxo complexes [Fe(II)(L)2(OH)](BF4) (1) and [Fe(III)(L)2(OH)2](BF4) (2) have been synthesized and characterized by X-ray diffraction and spectroscopic methods. The X-ray data suggest that the remarkable stability of the Fe-OH bond(s) in both compounds results from intermolecular hydrogen-bonding interactions between the hydroxo ligand(s) and the tertiary hydroxyl of the L ligands, which prevent further intermolecular reactions.     

Carbohydrate chain of ganglioside GM1 as a ligand: identification of the binding strategies of three 15 mer peptides and their divergence from the binding modes of growth-regulatory galectin-1 and cholera toxin

The branched pentasaccharide chain of ganglioside GM1 is a prominent cell surface ligand, for example, for cholera toxin or tumor growth-regulatory homodimeric galectins. This activity profile via protein recognition prompted us to examine the binding properties of peptides with this specificity. Our study provides insights into the mechanism of molecular interaction of this thus far unexplored size limit of the protein part. We used three pentadecapeptides in a combined approach of mass spectrometry, NMR spectroscopy and molecular modelling to analyze the ligand binding in solution. Availability of charged and hydrophobic functionalities affected the intramolecular flexibility of the peptides differently. Backfolding led to restrictions in two cases; the flexibility was not reduced significantly by association of the ligand in its energetically privileged conformations. Major contributions to the interaction energy arise from the sialic acid moiety contacting Arg/Lys residues and the N-terminal charge. Considerable involvement of stacking between the monovalent ligand and aromatic rings could not be detected. This carbohydrate binding strategy is similar to how an adenoviral fiber knob targets sialylated glycans. Rational manipulation for an affinity enhancement can now be directed to reduce the flexibility, exploit the potential for stacking and acquire the cross-linking capacity of the natural lectins by peptide attachment to a suitable scaffold.     

Distinction between surface hydroxyl and ether groups on boron-doped diamond electrodes using a chemical approach

While it is clearly established that oxidation of as-grown boron-doped diamond (BDD) interfaces results in the introduction of different surface oxygen functions such as ether, carbonyl and hydroxyl groups, there is no reported approach which can clearly distinguish between the different surface functions. For further surface functionalization, it is important to quantify the presence of each group on the diamond surface. In this paper, the presence and amount of surface hydroxyl groups is identified using esterification of the COH groups with trifluoroacetic acid. The presence of CF₃ group in the acid allowed the identification and estimation of the amount of surface hydroxyl groups using X-ray photoelectron spectroscopy (XPS).     

A bioinformatics approach to the development of immunoassays for specified risk material in canned meat products

A bioinformatics approach to developing antibodies to specific proteins has been evaluated for the production of antibodies to heat-processed specified risk tissues from ruminants (brain and eye tissue). The approach involved the identification of proteins specific to ruminant tissues by interrogation of the annotation fields within the Swissprot database. These protein sequences were then interrogated for peptide sequences that were unique to the protein. Peptides were selected that met these criteria as close as possible and that were also theoretically resistant to either pepsin or trypsin. The selected peptides were synthesised and used as immunogens to raise monoclonal antibodies. Antibodies specific for the synthetic peptides were raised to half of the selected peptides. These antibodies have each been incorporated into a competitive enzyme-linked immunosorbent assay (ELISA) and shown to be able to detect the heat-processed parent protein after digestion with either pepsin or trypsin. One antibody, specific for alpha crystallin peptide (from bovine eye tissue), was able to detect the peptide in canned meat products spiked with 10% eye tissue. These results, although preliminary in nature, show that bioinformatics in conjunction with enzyme digestion can be used to develop ELISA for proteins in high-temperature processed foods and demonstrate that the approach is worth further study.     

Concise Synthesis of [1,1′‐Biisoquinoline]‐4,4′‐diol via a Protecting Group Strategy and Its Application for Potential Liquid‐Crystalline Compounds

The [1,1′‐biisoquinoline]‐4,4′‐diol ( 4a ), which was obtained as hydrochloride 4a ?2 HCl in two steps starting from the methoxymethyl (MOM)‐protected 1‐chloroisoquinoline 8 (Scheme 3), opens access to further O‐functionalized biisoquinoline derivatives. Compound 4a ?2 HCl was esterified with 4‐(hexadecyloxy)benzoyl chloride ( 5b ) to give the corresponding diester 3b (Scheme 4), which could not be obtained by Ni‐mediated homocoupling of 6b (Scheme 2). The ether derivative 2b was accessible in good yield by reaction of 4a ?2 HCl with the respective alkyl bromide 9 under the conditions of Williamson etherification (Scheme 4). Slightly modified conditions were applied to the esterification of 4a ?2 HCl with galloyl chlorides 10a – h as well as etherification of 4a ?2 HCl with 6‐bromohexyl tris(alkyloxy)benzoates 11b , d – h and [(6‐bromohexyl)oxy]‐substituted pentakis(alkyloxy)triphenylenes 14a – c (Scheme 5). Despite the bulky substituents, the respective target 1,1′‐biisoquinolines 12, 13 , and 15 were isolated in 14–86% yield (Table).     

Water behavior in mesoporous materials as studied by NMR relaxometry

Water in mesoporous materials possessing a two-dimensional hexagonal structure has been studied by the variation of its NMR longitudinal relaxation time T(1) as a function of the static magnetic field value, or equivalently of the NMR measurement frequency. This technique, dubbed relaxometry, has been applied from 5 kHz (measurement frequency) up to 400 MHz with various instruments including a variable-field spectrometer operating between 8 and 90 MHz. Moreover, the range 0-5 kHz could be investigated by transverse relaxation, T(2) denoting the corresponding relaxation time, and relaxation in the rotating frame, T(1ρ) denoting the corresponding relaxation time. Measurements of proton relaxation rates (inverse of relaxation times) have been performed with H(2)O and HOD (residual protons of heavy water) at water volumes of 80%, 60%, and 40% relative to the porous volume. Comparison between H(2)O and HOD shows clearly that, above 1 MHz where both sets of data are superposed, relaxation is purely intermolecular and due to paramagnetic relaxation (dipolar interactions of water protons with unpaired electrons of paramagnetic entities). Below 1 MHz, it is possible to subtract the intermolecular contribution (given by HOD data) from H(2)O data so that one is left with intramolecular relaxation which is solely due to water reorientational motions. The analysis of these low-frequency data (in terms of Lorentzian functions) reveals two types of water within the pores: one interacting strongly with the surface and the other corresponding to a second layer. High-frequency data, which arise from paramagnetic relaxation, exhibit again two types of water. Due to their correlation times, one type is assigned to relatively free water within the pores while the other type corresponds to bulk (interparticular) water. Their proportions, given as a function of the volume fraction, are consistent with the above assignments.     

Aggregation behaviors of gemini nucleotide at the air-water interface and in solutions induced by adenine-uracil interaction

Cationic gemini surfactants having nucleotides as counterions (called nucleo-gemini hereafter) were synthesized and their aggregation behavior at air-water surfaces as well as in bulk solutions were studied. Fluid solutions of these nucleo-gemini surfactants show transitions to hydrogels upon addition of complementary nucleoside bases or other nucleo-gemini surfactants having complementary bases as counterions. The FTIR-ATR measurements show that the carboxylate groups of uridine form hydrogen bonds with the amine groups of adenosine. The aggregation behavior was also confirmed at the air-water interface by Brewster angle microscopy as well as surface pressure measurements; the monolayer of a gemini nucleotide was observed to undergo a transition to multilayers when nucleosides with complementary bases were added into the subphase. Isotherm curves of surface pressure monitored in parallel show a decrease in molecular area upon addition of such nucleosides.