First application of the depth profile of silica species as a tracer by fast-atom bombardment mass spectrometry: investigation of the circulation of seawater and silica uptake by diatoms.

Silica, represented by SiO(2), is the general name for the compounds composed of Si, O and H with their derivative complexes. Silica forms various chemical species in aquatic solutions, such as a monomer (Si(OH)(3)O(-)), dimer (Si(2)(OH)(5)O(2) (-)), and others. These species are known to vary in their relative abundances in solution depending on the chemical and physical conditions. Silica species dissolved in seawater have been examined by fast-atom bombardment mass spectrometry (FAB-MS) to elucidate the behavior of silica and its circulation as a novel tracer reflecting the chemical and physical conditions of seawater and the bioactivity of diatoms, which take up silica. In the seawater of Tokyo Bay, silica species such as [Si(OH)(2)O(2)Na](-) ([monomer-Na](-)), [Si(2)(OH)(5)O(2)](-) ([dimer](-)), [Si(2)(OH)(4)O(3)Na](-) ([dimer-Na](-)), [Si(4)(OH)(7)O(5)](-) ([cyclic tetramer](-)), [Si(4)(OH)(6)O(6)Na](-) ([cyclic tetramer-Na](-)), [Si(4)(OH)(9)O(4)](-) ([linear tetramer](-)) and [Si(4)(OH)(8)O(5)Na](-) ([linear tetramer-Na](-)) were observed and assigned by FAB-MS. To investigate the suitability of silica species as a tracer, the relative peak intensity ratios of silica species observed in the mass spectra, i.e. the profiles of the ratio of the linear tetramer to the cyclic tetramer (m/z 329/311) and the ratio of the dimer to the cyclic tetramer (m/z 173/311) against depth, were examined to determine the annual changes and reproducibility of the depth profiles. In particular, the depth profile of the relative ratio of the linear tetramer to the cyclic tetramer, 329/311, exhibits critical changes depending on the seawater budget. These changes in the relative ratios were identified by an experiment involving a simple sodium chloride solution system. Our measurement is expected to elucidate the dynamics of silica and its role as 'food' for diatoms, and we showed that speciation using mass spectrometry is a powerful tool for examining elemental behavior in nature and environmental changes. Our results suggest that a silica tracer is useful for investigating the behavior of seawater in small coastal regions and the uptake of silica by diatoms.

Caenorhabditis elegans mutants having altered preference of chemotaxis behavior during simultaneous presentation of two chemoattractants.

Upon presentation of two distinct chemoattractants such as sodium acetate and diacetyl simultaneously, the nematode Caenorhabditis elegans was preferentially attracted by one of these chemoattractants. We isolated two mutants having altered preference of chemotaxis behavior toward simultaneous presentation of sodium acetate and diacetyl. The chep-1(qr1) (CHEmosensory Preference) mutant preferred sodium acetate to diacetyl, while the chep-2(qr2) mutant preferred diacetyl to sodium acetate in simultaneous presentation of these chemoattractants. The chemotaxis behavior of chep-2(qr2) mutant in simultaneous presentation suggests a function of chep-2 gene products within the chemosensory informational integration pathway as well as in the chemosensory pathway.

Effect of simultaneous presentation of multiple attractants on chemotactic response of the nematode Caenorhabditis elegans.

Chemotactic behaviors of the nematode, Caenorhabditis elegans in response to chemical attractants, such as water-soluble sodium acetate and an odorant diacetyl, which were sensed by different sensory neurons, were investigated using various concentrations of these chemical attractants. In the presence of only sodium acetate attractant, the fraction of animals that were roaming around the outside of the attractant and original locations correlated negatively with the chemotaxis index for sodium acetate (P < 0.01). In contrast, the fraction of animals that remained in the original location correlated negatively with the chemotaxis index in the presence of only diacetyl attractant (P < 0.05). These results indicate that the manner of chemotaxis responses differs between sodium acetate and diacetyl. In order to investigate the effect of multiple attractants on chemotactic behaviors, the chemotactic responses to simultaneous presentation of sodium acetate and diacetyl were examined. The fraction of animals that gathered at the 0.7 M sodium acetate location was greater than that at the 0.1% diacetyl location in the presence of both attractants (P < 0.05), although the chemotaxis indexes for 0.7 M sodium acetate and 0.1% diacetyl were similar in the presence of a single attractant. On the other hand, the fraction of animals that gathered at the 0.02% diacetyl location was greater than that at the 0.1M sodium acetate location in the presence of both attractants (P < 0.05), although the chemotaxis indexes for 0.02% diacetyl and 0.1M sodium acetate were similar in the presence of a single attractant. These results suggest the existence of excitatory and/or inhibitory connections in the neuronal circuit for attractant selection, and that the efficacy of these connections may change according to the concentrations of both attractants.

Improvement of DNA vaccine immunogenicity by a dual antigen expression system.

This study examined whether increased antigen expression resulted in enhanced antigen-specific immune responses in the context of DNA vaccines. To increase antigen expression, two copies of antigen expression cassettes were arranged in a plasmid pDX. BALB/c mice were intramuscularly immunized with various constructs that express influenza antigens and analysed for DNA-raised immunity. The plasmid pDX that expresses two copies of the antigen gene induced stronger antigen-specific immune responses than the plasmid pGA which expresses single antigen gene. To explore the in vivo transgene expression by pDX and pGA, luciferase activity was measured in the muscles transduced with luciferase expression plasmids. The pDX expressing two copies of luciferase induced the highest luciferase activity, which corresponded to the results from vaccination. We concluded that increasing the number of antigen expression cassettes in a vaccine construct improved antigen expression in the transduced tissue, which induced stronger DNA-raised immune responses.

Adjuvant effect of multi-CpG motifs on an HIV-1 DNA vaccine.

Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG motifs trigger an immune response characterized by the activation of B cells, NK cells and monocytes/macrophages. Based on evidence that the immunogenicity of DNA vaccines can be augmented by the addition of CpG motifs, 5-20 additional CpG motifs were cloned into a pUC-derived plasmid. Treating bone-marrow derived dendritic cells (BM-DCs) with CpG-enriched plasmids in vitro boosted their expressions of MHC class II molecules, the CD40 and CD86 activation markers. Co-administering the CpG-enriched plasmids with a DNA vaccine encoding the envelope glycoprotein of HIV to BALB/c mice significantly increased HIV-specific cell mediated and humoral immunity. A significant boost was observed when the CpG plasmid was administered either 2 or 4 days after DNA vaccination. Plasmids containing 20 CpG copies were the most effective immune enhancers both in vitro and in vivo. These results suggest that plasmids containing multiple CpG motifs may improve the immunogenicity of DNA vaccines.