Complete Genome Sequence of Human Respiratory Syncytial Virus from Lanzhou, China.

A complete genome of human respiratory syncytial virus was sequenced and analyzed. Phylogenetic analysis showed that the full-length human respiratory syncytial virus (HRSV) genome sequence belongs to gene type NA1. We sequenced the genome in order to create the full-length cDNA infectious clone and develop vaccines against HRSV.

OsLOL1, a C2C2-type zinc finger protein, interacts with OsbZIP58 to promote seed germination through the modulation of gibberellin biosynthesis in Oryza sativa.

Seed germination is a key developmental process in the plant life cycle that is influenced by various environmental cues and phytohormones through gene expression and a series of metabolism pathways. In the present study, we investigated a C2C2-type finger protein, OsLOL1, which promotes gibberellin (GA) biosynthesis and affects seed germination in Oryza sativa (rice). We used OsLOL1 antisense and sense transgenic lines to explore OsLOL1 functions. Seed germination timing in antisense plants was restored to wild type when exogenous GA3 was applied. The reduced expression of the GA biosynthesis gene OsKO2 and the accumulation of ent-kaurene were observed during germination in antisense plants. Based on yeast two-hybrid and firefly luciferase complementation analyses, OsLOL1 interacted with the basic leucine zipper protein OsbZIP58. The results from electrophoretic mobility shift and dual-luciferase reporter assays showed that OsbZIP58 binds the G-box cis-element of the OsKO2 promoter and activates LUC reporter gene expression, and that interaction between OsLOL1 and OsbZIP58 activates OsKO2 gene expression. In addition, OsLOL1 decreased SOD1 gene expression and accelerated programmed cell death (PCD) in the aleurone layer of rice grains. These findings demonstrate that the interaction between OsLOL1 and OsbZIP58 influences GA biosynthesis through the activation of OsKO2 via OsbZIP58, thereby stimulating aleurone PCD and seed germination.

Overexpression of a cotton cyclophilin gene (GhCyp1) in transgenic tobacco plants confers dual tolerance to salt stress and Pseudomonas syringae pv. tabaci infection.

The full-length cDNA of a cyclophilin-like gene was cloned from Gossypium hirsutum using rapid amplification of cDNA ends and was designated as GhCyp1, a member of the immunophilin protein family. GhCyp1 expression level was higher in roots and stems than in other tissues of cotton, as determined by real-time reverse transcription polymerase chain reaction (RT-PCR). To characterize the GhCyp1 gene, tobacco (Nicotiana tabacum) was transformed via Agrobacterium tumefaciens with a vector to express the gene under the control of a strong constitutive promoter, CaMV35S (Cauliflower Mosaic Virus). Based on analyses of tolerance to salinity stress and Pseudomonas syringae pv. tabaci (Pst) infection, the overexpression of GhCyp1 in transgenic plants conferred higher tolerance to salt stress and Pst infection compared with control plants. Therefore, we suggest that GhCyp1 may be a suitable candidate gene to produce transgenic plants with tolerance to abiotic and biotic stresses.

Transmembrane delivery of aggregated [Gd@C82(OH)22]n nanoparticles.

We investigated whether multi-hydroxyl metallofullerenes can penetrate into erythrocyte and whether this potential transmembrane delivery requires aggregated nanostructure of these particles. The metal atom encapsulated in metallofullerenes was used as a quantitative marker to investigate body distribution of aggregated nanoparticles in cytomembrane and cytoplasm. Image of atomic force microscopy (AFM) and assay of inductively coupled plasma-mass spectrometry (ICP-MS) suggested that aggregated [Gd@C82(OH)22]n particles traversed through cytomembrane into cytoplasm. Aggregated Gd nanostructure belonged to small sphere with average diameter of 22.4 +/- 0.5 nm. For pristine Gd@C82 molecule, due to the electron donation from Gd atom, the distribution of electrons on the surface of carbon cage was localized. The electrophilic additive reaction of polyhydroxyl on the surface of Gd@C82 was directly affected by local distribution of electrons. This resulted in local distribution of hydroxyls on the surface of Gd nanoparticles. Local distribution of hydroxyls brought about polar and nonpolar domains on particle surface, which induced Gd@C82(OH)22 to be amphiphilic molecule with due hydrophilic and hydrophobic properties. Amphiphilic properties of these molecules promoted their mutual aggregation in water. In the process of aggregation, amphiphilic properties of aggregated nanoparticles were well maintained, and besides, hydrophilic and hydrophobic domains were also regularly distributed on the surface of [Gd@C82(OH)22]n particles. The amphiphilic nanoparticles attached externally to cytomembrane of erythrocyte might be effectively driven by hydrophobic effect when they directly contacted cytomembrane of erythrocyte. The number of [Gd@C82(OH)22]n nanoparticles attached to cytomembrane reached up to a certain critical threshold, a significant curvature tension of membrane would occur. The shape of cell was accordingly changed. Increased membrane tension triggered the sudden opening of specific pores as a result of cytmembrane response to nanoparitcle effect and the [Gd@C82(OH)22]n nanoparticles gained entry to cell via these pores. This process was biologically independent of caveolar-mediated endocytosis or transportation pathway via ion channels.

Induction of chromosomal inversion by integration of T-DNA in the rice genome.

Transfer DNA (T-DNA) of Agrobacterium tumefaciens integration in the plant genome may lead to rearrangements of host plant chromosomal fragments, including inversions. However, there is very little information concerning the inversion. The present study reports a transgenic rice line selected from a T-DNA tagged population, which displays a semi-dwarf phenotype. Molecular analysis of this mutant indicated an insertion of two tandem copies of T-DNA into a locus on the rice genome in a head to tail mode. This insertion of T-DNA resulted in the inversion of a 4.9 Mb chromosomal segment. Results of sequence analysis suggest that the chromosomal inversion resulted from the insertion of T-DNA with the help of sequence microhomology between insertion region of T-DNA and target sequence of the host plant.

Phage M13KO7 detection with biosensor based on imaging ellipsometry and AFM microscopic confirmation.

A rapid detection and identification of pathogens is important for minimizing transfer and spread of disease. A label-free and multiplex biosensor based on imaging ellipsometry (BIE) had been developed for the detection of phage M13KO7. The surface of silicon wafer is modified with aldehyde, and proteins can be patterned homogeneously and simultaneously on the surface of silicon wafer in an array format by a microfluidic system. Avidin is immobilized on the surface for biotin-anti-M13 immobilization by means of interaction between avidin and biotin, which will serve as ligand against phage M13KO7. Phages M13KO7 are specifically captured by the ligand when phage M13KO7 solution passes over the surface, resulting in a significant increase of mass surface concentration of the anti-M13 binding phage M13KO7 layer, which could be detected by imaging ellipsometry with a sensitivity of 10(9)pfu/ml. Moreover, atomic force microscopy is also used to confirm the fact that phage M13KO7 has been directly captured by ligands on the surface. It indicates that BIE is competent for direct detection of phage M13KO7 and has potential in the field of virus detection.

A comparative histologic study of rapid three-dimensional zygomatic suture expansion osteogenesis.

OBJECTIVE: To explore the histologic changes in zygomatic suture with 3-dimensional (3D) zygomatic suture expansion osteogenesis (SEO). STUDY DESIGN: The zygomatic bones were drawn by 3D external expansion appliance, and sutures of the zygomatic were extended. Biopsy specimens of the zygomatic bone were collected after 1, 3, 5, and 8 weeks. Each specimen was stained with Triplex staining as well as hematoxylin and eosin (HE), and the histologic changes were evaluated compared with the control section. RESULTS: With the 1-week study group, there were visible fibroblasts, osteoblasts, and capillary vessels in the expanded suture tissues. The fibers were connected to the sides of suture in an orderly way. In the 3-week group, active bone formation can be seen in expanded sides. The bone trabeculae were matured and oriented in the direction of distraction in the expanded sides of suture and most of them were collagen fibers; but the tissues of both sides of expanded suture were not in order. New woven bones were found in the histologic sections of the 5-week group. In the 8-week group, reticular and elastic fibers could not be observed, and bones were formed completely. Compared with the histologic examination of the same tissue section and its change in different periods of the biopsy specimen with Triplex staining and HE staining, the method of Triplex staining showed more clearly in collagen, reticular, and elastic fiber examination. The superiority of HE staining was in showing fibrous tissue, shape of cells, and bone formation in various degrees of maturation in rapid SEO. It is suggested that the 2 staining methods might be combined in distinguishing the collagen as well as reticular fibers from fibrin in the same section simultaneously. CONCLUSIONS: Collagen fibers and new bones were observed to form rapidly in expanded sides with direct SEO. Continual affluent blood supply, integral structure of periosteum and synostoses in the suture, and rapid uniform calcification of the whole new tissues are the histologic characteristics of SEO.

Boric acid and salinity effects on maize roots. Response of aquaporins ZmPIP1 and ZmPIP2, and plasma membrane H+-ATPase, in relation to water and nutrient uptake.

Under saline conditions, an optimal cell water balance, possibly mediated by aquaporins, is important to maintain the whole-plant water status. Furthermore, excessive accumulation of boric acid in the soil solution can be observed in saline soils. In this work, the interaction between salinity and excess boron with respect to the root hydraulic conductance (L(0)), abundance of aquaporins (ZmPIP1 and ZmPIP2), ATPase activity and root sap nutrient content, in the highly boron- and salt-tolerant Zea mays L. cv. amylacea, was evaluated. A downregulation of root ZmPIP1 and ZmPIP2 aquaporin contents were observed in NaCl-treated plants in agreement with the L(0) measurements. However, in the H3BO3-treated plants differences in the ZmPIP1 and ZmPIP2 abundance were observed. The ATPase activity was related directly to the amount of ATPase protein and Na+ concentration in the roots, for which an increase in NaCl- and H3BO3+ NaCl-treated plants was observed with respect to untreated and H3BO3-treated plants. Although nutrient imbalance may result from the effect of salinity or H3BO3 alone, an ameliorative effect was observed when both treatments were applied together. In conclusion, our results suggest that under salt stress, the activity of specific membrane components can be influenced directly by boric acid, regulating the functions of certain aquaporin isoforms and ATPase as possible components of the salinity tolerance mechanism.

Topography and functional information of plasma membrane.

By using atomic force microscope (AFM), the topography and function of the plasmalemma surface of the isolated protoplasts from winter wheat mesophyll cells were observed, and compared with dead protoplasts induced by dehydrating stress. The observational results revealed that the plasma membrane of living protoplasts was in a state of polarization. Lipid layers of different cells and membrane areas exhibited distinct active states. The surfaces of plasma membranes were unequal, and were characterized of regionalisation. In addition, lattice structures were visualized in some regions of the membrane surface. These typical structures were assumed to be lipid molecular complexes, which were measured to be 15.8+/-0.09 nm in diameter and 1.9+/-0.3 nm in height. Both two-dimensional and three-dimensional imaging showed that the plasmalemma surfaces of winter wheat protoplasts were covered with numerous protruding particles. In order to determine the chemical nature of the protruding particles, living protoplasts were treated by proteolytic enzyme. Under the effect of enzyme, large particles became relatively looser, resulting that their width was increased and their height decreased. The results demonstrated that these particles were likely to be of protein nature. These protein particles at plasmalemma surface were different in size and unequal in distribution. The diameter of large protein particles ranged from 200 to 440 nm, with a central micropore, and the apparent height of them was found to vary from 12 to 40 nm. The diameter of mid-sized protein particles was between 40-60 nm, and a range of 1.8-5 nm was given for the apparent height of them. As for small protein particles, obtained values were 12-40 nm for their diameter and 0.7-2.2 nm for height. Some invaginated pits were also observed at the plasma membrane. They were formed by the endocytosis of protoplast. Distribution density of them at plasmalemma was about 16 pits per 15 microm(2). According to their size, we classified the invaginated pits into two types--larger pits measuring 139 nm in diameter and 7.2 nm in depth, and smaller pits measuring 96 nm in diameter and 2.3 nm in depth. On dehydration-induced dead protoplasts, the degree of polarization of plasma membranes decreased. Lipid molecular layers appeared relatively smooth, and the quantity of integral proteins reduced a lot. Invaginated pits were still detectable at the membrane surface, but due to dehydration-induced protoplast contraction, the orifice diameter of pits reduced, and their depth increased. Larger pits averagely measuring 47.4 nm in diameter and 31.9 nm in depth, and smaller pits measuring 26.5 nm in diameter and 43 nm in depth at average. The measured thickness of plasma membranes of mesophyll cells from winter wheat examined by AFM was 6.6-9.8 nm, thicker in regions covered with proteins.

Aggregation studies of the water-soluble gadofullerene magnetic resonance imaging contrast agent: [Gd@C82O6(OH)16(NHCH2CH2COOH)8]x.

The aggregation behavior of the newly synthesized gadofullerene magnetic resonance imaging (MRI) contrast agent, i.e., Gd@C(82)O(6)(OH)(16)(NHCH(2)CH(2)COOH)(8) (abbreviated as AAD-EMF), was studied in detail by dynamic light scattering, scanning electron microscopy, T(1)-weighted magnetic resonance, and atomic force microscopy. It was revealed that the AAD-EMF aggregation in aqueous solution is pH-dependent. At pH 2, the AAD-EMF first self-assemble to form ca. 30 nm small clusters, and then dozens of the small clusters further aggregate to form large grapelike particles. At pH 7, the aggregates are also ca. 30 nm small clusters, but they are hard to further aggregate except for forming some cluster dimers or trimers, so AAD-EMF aggregates have a narrow size distribution by this time. At pH 9, the AAD-EMF aggregations cover a large range of continuous hydrodynamic diameters from 30 to 2000 nm. On the basis of the above observations, the aggregating mechanism of AAD-EMF under different pH values was proposed by concurrently considering the hydrogen-bonding effect and the dipolar interactions between AAD-EMF.

Antioxidative function and biodistribution of [Gd@C82(OH)22]n nanoparticles in tumor-bearing mice.

Oxidative stress is considered to be one of the important mechanisms involved in carcinogenesis. In our previous study, gadolinium endohedral metallofullerenol ([Gd@C82(OH)22]n nanoparticles) have shown high inhibitory activity on hepatoma cell (H22) growth in mice. To explore the antioxidative functions of nanoparticles, we investigated the biodistribution of [Gd@C82(OH)22]n nanoparticles, the changes of blood coagulation profiles, the metabolism of reactive oxygen species (ROS) in the tumor-bearing mice as well as the possible relationships between nanoparticles treatment and ROS production in this paper. The activities of hepatic superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST) and catalase (CAT) as well as the levels of reduced glutathione (GSH), protein-bound thiols and malondialdehyde (MDA) were compared between the tumor-bearing mice and normal mice. Transplanted tumors were grown in mice by subcutaneous injection of murine hepatoma cells in the mice. The comparison of the above parameters between nanoparticles and cyclophosphamide (CTX) therapy were also investigated. [Gd@C82(OH)22]n administration can efficiently restore the damaged liver and kidney of the tumor-bearing mice. All the activities of enzymes and other parameters related to oxidative stress were reduced after [Gd@C82(OH)22]n treatment and tended closely to the normal levels. The results suggest that [Gd@C82(OH)22]n nanoparticle treatment could regulate ROS production in vivo.

Acute toxicological effects of copper nanoparticles in vivo.

To assess the toxicity of copper nanoparticles (23.5 nm) in vivo, LD(50), morphological changes, pathological examinations and blood biochemical indexes of experimental mice are studied comparatively with micro-copper particles (17 microm) and cupric ions (CuCl(2).2H(2)O). The LD(50) for the nano-, micro-copper particles and cupric ions exposed to mice via oral gavage are 413, >5000 and 110 mg/kg body weight, respectively. The toxicity classes of nano and ionic copper particles both are class 3 (moderately toxic), and micro-copper is class 5 (practically non-toxic) of Hodge and Sterner Scale. Kidney, liver and spleen are found to be target organs of nano-copper particles. Nanoparticles induce gravely toxicological effects and heavy injuries on kidney, liver and spleen of experimental mice, but micro-copper particles do not, on mass basis. Results indicate a gender dependent feature of nanotoxicity. Several factors such as huge specific surface area, ultrahigh reactivity, exceeding consumption of H(+), etc. that likely cause the grave nanotoxicity observed in vivo are discussed.

Differential responses of maize MIP genes to salt stress and ABA.

Salt stress is known to reduce root hydraulic conductivity and growth. To examine a concomitant regulation of aquaporins, the expression of the maize MIP gene family in response to NaCl was analysed by DNA array hybridization. Plants responded differentially to 100 versus 200 mM NaCl treatments. Leaf water content was reduced rapidly and persistently after the application of 200 mM NaCl in contrast to 100 mM NaCl. Endogenous ABA strongly accumulated in roots after 2 h; it remained at a highly elevated level for 48 h after the addition of 200 mM NaCl, but rapidly declined in plants treated with 100 mM NaCl, indicating an early recovery from water deficit. Interestingly, 2 h after the addition of 100 mM NaCl, when maize regained the osmotic potential allowing water uptake, three highly expressed, specific isoforms ZmPIP1;1, ZmPIP1;5, and ZmPIP2;4 were transiently induced. They were preferentially transcribed in the outer root tissue suggesting a role in cellular water transport. None of the ZmTIP genes was altered. By contrast, after the addition of 200 mM NaCl these responses were missing. Instead, multiple ZmPIP and ZmTIP genes were repressed by 200 mM NaCl after 24 h. After 48 h, deregulations were overridden in both cases indicating homeostasis. ABA (1 muM) exogenously applied to the roots transiently induced ZmPIP2;4 similar to 100 mM NaCl as well as ZmPIP1;2. Thus, the early induction of ZmPIP2;4 by NaCl may be mediated by ABA. Previously, an increase in root hydraulic conductivity had been observed upon ABA application. By contrast, 100 muM ABA led to a complete, possibly non-specific repression of all detected ZmPIP and ZmTIP genes after 24 h.

Ni-Pt multilayered nanowire arrays with enhanced coercivity and high remanence ratio.

Highly ordered Ni-Pt multilayered nanowire arrays have been fabricated using a porous anodic aluminum oxide (AAO) template by pulse electrodeposition. The cylindrical Ni nanoparticles with different lengths and diameters in these arrays were characterized by transmission electron microscope (TEM) and alternating-gradient magnetometer (AGM) measurements. Magnetization measurements revealed that an array of such nanowires with 20-nm diameters has an enhanced coercivity (ca. 1169 Oe) and a high remanence ratio (ca. 0.96).

Characterization of surface property of poly(lactide-co-glycolide) after oxygen plasma treatment.

In this study, poly (lactide-co-glycolide) (PLGA) films were treated by oxygen plasma. The surface structure, topography and surface chemistry of treated PLGA films were characterized by contact angle measurement, scanning electron microscope observation, atomic force microscopy, and X-ray photoelectron spectrum analysis. The cell affinity of the oxygen plasma treated films was evaluated under dynamic conditions by Parallel Plate Flow Chamber (PPFC). The results showed that the hydrophilicity increased greatly after oxygen plasma treatment. High quantities of -C-O groups, such as hydroxyl and peroxyl groups could be incorporated into the surface of PLGA (70/30) by controlling appropriate plasma treatment conditions. Moreover, the oxygen plasma treatment resulted in formation of peaks and valleys on the sample surfaces, and the roughness increased with treatment time. Cells stretched very well and the ability to endure the shear stress was improved greatly after the PLGA (70/30) was modified by appropriate plasma treatment, i.e. under 50W for 2 or 10 min. However, when the treatment time was increased to 20 min, the percentage of adherent cells on the roughest surface decreased because the content of polar groups incorporated onto the surface decreased. The results showed that improved cell adhesion was attributed to the combination of surface chemistry and surface morphology of PLGA during plasma etching.

Specific aptamer-protein interaction studied by atomic force microscopy.

Aptamers are a new class of synthetic DNA/RNA oligonucleotides generated from in vitro selection to selectively bind with various molecules. Due to their molecular recognition capability for proteins, aptamers are becoming promising reagents in protein detection and new drug development. In this study, the specific interaction between the protein immunoglobulin E (IgE) and its 37-nt aptamer has been measured directly by atomic force microscopy. The single-molecule unbinding force between IgE and the aptamer is determined using the Poisson statistical method. The individual unbinding force between IgE and its monoclonal antibody has also been obtained and compared to that between IgE and the aptamer. The results reveal the high affinity of the aptamer to protein, which could match or even surpass that of the antibody to its antigen.