Direct observation of chemokine receptors 5 on T-lymphocyte cell surfaces using fluorescent metal nanoprobes 2: Approximation of CCR5 populations

Metal nanoparticle probes were used as molecular imaging agents to detect the expression levels and spatial distributions of the CCR5 receptors on the cell surfaces. Alexa Fluor 647-labeled anti-CCR5 monoclonal antibodies (mAbs) were covalently bound to 20 nm silver nanoparticles to synthesize the mAb–metal complexes. We measured the single nanoparticle emission of the mAb–metal complexes, showing that the complexes displayed enhanced intensities and reduced lifetimes in comparison with the metal-free mAbs. Six HeLa cell lines with various CCR5 expressions were incubated with the mAb–metal complexes for the target-specific binding to the cell surfaces. Fluorescence cell images were recorded on a time-resolved confocal microscope. The collected images expressed clear CCR5 expression-dependent optical properties. Two regression curves were obtained on the basis of the emission intensity and lifetime over the entire cell images against the number of the CCR5 expression on the cells. The emission from the single mAb–metal complexes could be distinctly identified from the cellular autofluorescence on the cell images. The CCR5 spatial distributions on the cells were analyzed on the cell images and showed that the low-expression cells have the CCR5 receptors as individuals or small clusters but the high expression cells have them as the dense and discrete clusters on the cell surfaces.     

Hypoxia induces beta-amyloid in association with death of RGC-5 cells in culture

Beta-amyloid (Aβ) derived from amyloid precursor protein (APP) has been associated with retinal degeneration in Alzheimer’s disease (AD) and glaucoma. This study examined whether hypoxia exposure induces Aβ accumulation in RGC-5 cells. While levels of APP mRNA and protein significantly increased in the cells, elevated abundance of Aβ was also observed in cells and culture medium between 12 or 24 and 48 h after 5% O₂ hypoxia treatment. Additionally, there is a close relationship between induction of APP and Aβ and intracellular accumulation of ROS along with loss of mitochondrial membrane potential followed by the death of RGC-5 cells in culture under hypoxia. These results suggest a possible involvement of APP and Aβ in the death of RGCs challenged by hypoxia.     

Lipid interacting regions in phosphate stress glycosyltransferase atDGD2 from Arabidopsis thaliana

Membrane lipid glycosyltransferases (GTs) in plants are enzymes that regulate the levels of the non-bilayer prone monogalactosyldiacylglycerol (GalDAG) and the bilayer-forming digalactosyldiacylglycerol (GalGalDAG). The relative amounts of these lipids affect membrane properties such as curvature and lateral stress. During phosphate shortage, phosphate is rescued by replacing phospholipids with GalGalDAG. The glycolsyltransferase enzyme in Arabidopsis thaliana responsible for this, atDGD2, senses the bilayer properties and interacts with the membrane in a monotopic manner. To understand the parameters that govern this interaction, we have identified several possible lipid-interacting sites in the protein and studied these by biophysical techniques. We have developed a multivariate discrimination algorithm that correctly predicts the regions in the protein that interact with lipids, and the interactions were confirmed by a variety of biophysical techniques. We show by bioinformatic methods and circular dichroism (CD), fluorescence, and NMR spectroscopic techniques that two regions are prone to interact with lipids in a surface-charge dependent way. Both of these regions contain Trp residues, but here charge appears to be the dominating feature governing the interaction. The sequence corresponding to residues 227-245 in the protein is seen to be able to adapt its structure according to the surface-charge density of a bilayer. All results indicate that this region interacts specifically with lipid molecules and that a second region in the protein, corresponding to residues 130-148, also interacts with the bilayer. On the basis of this, and sequence charge features in the immediate environment of S227-245, a response model for the interaction of atDGD2 with the membrane bilayer interface is proposed.     

Complete genome sequence of Paenibacillus polymyxa SC2, a strain of plant growth-promoting Rhizobacterium with broad-spectrum antimicrobial activity

Paenibacillus polymyxa SC2 is an important plant growth-promoting rhizobacterium (PGPR). Here, we report the complete genome sequence of P. polymyxa SC2. Multiple sets of functional genes have been found in the genome. As far as we know, this is the first complete genome sequence of Paenibacillus polymyxa.     

Temporal and spatial variability of chlorophyll <Emphasis Type="Italic">a</Emphasis> concentration in Lake Taihu using MODIS time-series data

In order to predict the distribution of chlorophyll a synoptically in Lake Taihu from 2006 to 2008, a common empirical algorithm was developed to relate time series chlorophyll a concentrations in the lake to reflectance derived as a function of band 2 MODIS data (r ² = 0.907, n = 145) using time series from 2005. The empirical model was further validated with chlorophyll a data from a 2008 to 2009 dataset, with RMSE < 7.48 μg l⁻¹ and r ² = 0.978. The seasonal and inter-annual variability of the surface chlorophyll a concentration from 2006 to 2008 was then examined using Empirical Orthogonal Function (EOF) analysis. The results revealed that the first four modes were significant, explaining 54.0% of the total chlorophyll a variance, and indicated that during the summer, algal blooms always occur in the northern bays, Meiliang Bay and Gonghu Bay, while they occur along the southwestern lakeshore during early summer, fall, and even early winter. The inter-annual variance analysis showed that the duration of algal blooms was from April to December of 2007, which was different from the bloom periods in 2006 and 2008. The results of EOF analysis show its potential for long-term integrated lake monitoring, not only in Lake Taihu but also in other large lakes threatened by accelerating eutrophication.     

Involvement of auxin and nitric oxide in plant Cd-stress responses

Cadmium (Cd) toxicity inhibited the seedling growth while inducing the occurrences of lateral roots (LR) and adventitious roots (AR). Further study indicated that auxin and nitric oxide (NO) are involved in the processes. In this study, we chose model plant Arabidopsis thaliana and Cd-hyperaccumulator Solanum nigrum as material to examine the involvement of Cd-induced auxin redistribution in NO accumulation in plants and the effect of NO on Cd accumulation. For this aim, the histochemical staining, NO fluorescence probe (DAF-2DA) detections combined with the pharmacological study were used in this study. By using DR5:GUS staining analysis combined with NO fluorescence probe (DAF-2DA) detection, we found that Cd-induced NO accumulation is at least partly due to auxin redistribution in plants exposure to Cd. Supplementation with SNP donor S-nitrosoglutathione (GSNO) increased the number of LR and AR. In contrast, NO-scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide (cPTIO) reversed the effects of NO on modulating root system architecture and Cd accumulation. These results suggest that manipulation of the NO level is an effective approach to improve Cd tolerance in plants by modulating the development of LR and AR, and provide insights into novel strategies for phytoremediation.     

The complete mitochondrial genome of the marbled rockfish Sebastiscus marmoratus (Scorpaeniformes, Scorpaenidae): genome characterization and phylogenetic considerations

The complete mitochondrial genome sequence of the marbled rockfish Sebastiscus marmoratus (Scorpaeniformes, Scorpaenidae) was determined and phylogenetic analysis was conducted to elucidate the evolutionary relationship of the marbled rockfish with other Sebastinae species. This mitochondrial genome, consisting of 17301 bp, is highly similar to that of most other vertebrates, containing the same gene order and an identical number of genes or regions, including 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and one putative control region. Most of the genes are encoded on the H-strand, while the ND6 and seven tRNA genes (for Gln, Ala, Asn, Tyr, Ser (UCA), Glu, and Pro) are encoded on the L-strand. The reading frame of two pairs of genes overlapped on the same strand (the ATPase 8 and 6 genes overlapped by ten nucleotides; ND4L and ND4 genes overlapped by seven nucleotides). The possibly nonfunctional light-strand replication origin folded into a typical stem-loop secondary structure and a conserved motif (5'-GCCGG-3') was found at the base of the stem within the tRNA(Cys) gene. An extent termination-associated sequence (ETAS) and conserved sequence blocks (CSB) were identified in the control region, except for CSB-1; unusual long tandem repeats were found at the 3' end of the control region. Phylogenetic analyses supported the view that Sebastinae comprises four genera (Sebates, Hozukius, Helicolenus, and Sebasticus).