Synthesis and characterization of dual-functionalized core-shell fluorescent microspheres for bioconjugation and cellular delivery

The efficient transport of micron-sized beads into cells, via a non-endocytosis mediated mechanism, has only recently been described. As such there is considerable scope for optimization and exploitation of this procedure to enable imaging and sensing applications to be realized. Herein, we report the design, synthesis and characterization of fluorescent microsphere-based cellular delivery agents that can also carry biological cargoes. These core-shell polymer microspheres possess two distinct chemical environments; the core is hydrophobic and can be labeled with fluorescent dye, to permit visual tracking of the microsphere during and after cellular delivery, whilst the outer shell renders the external surfaces of the microspheres hydrophilic, thus facilitating both bioconjugation and cellular compatibility. Cross-linked core particles were prepared in a dispersion polymerization reaction employing styrene, divinylbenzene and a thiol-functionalized co-monomer. These core particles were then shelled in a seeded emulsion polymerization reaction, employing styrene, divinylbenzene and methacrylic acid, to generate orthogonally functionalized core-shell microspheres which were internally labeled via the core thiol moieties through reaction with a thiol reactive dye (DY630-maleimide). Following internal labeling, bioconjugation of green fluorescent protein (GFP) to their carboxyl-functionalized surfaces was successfully accomplished using standard coupling protocols. The resultant dual-labeled microspheres were visualized by both of the fully resolvable fluorescence emissions of their cores (DY630) and shells (GFP). In vitro cellular uptake of these microspheres by HeLa cells was demonstrated conventionally by fluorescence-based flow cytometry, whilst MTT assays demonstrated that 92% of HeLa cells remained viable after uptake. Due to their size and surface functionalities, these far-red-labeled microspheres are ideal candidates for in vitro, cellular delivery of proteins, as described in the accompanying paper.

Highly thermally responsive hybrid in-fiber slab waveguide grating structure with a plastic core and a silica cladding

A 1.2(height)×125(depth)×500(length) micro-slot was engraved along a fiber Bragg grating by chemically assisted femtosecond laser processing. By filling epoxy and UV-curing, waveguide with plastic-core and silica-cladding was created, presenting high thermal responding coefficient of 211pm/°C.

Realisation of an effective dual-parameter sensor employing a single fibre Bragg grating structure

A novel technology for simultaneous and independent measurement of dual parameters is proposed and experimented. The length of a single fibre Bragg grating (FBG) is divided into two parts. The temperature variation and another measurand can be measured independently and simultaneously, and the thermal effect can be erased with great ease. © 2005 Elsevier Ltd. All rights reserved.

Highly thermally responsive hybrid in-fiber sSlab waveguide grating structure with a plastic core and a silica cladding

A 1.2(height)×125(depth)×500(length) micro-slot was engraved along a fiber Bragg grating by chemically assisted femtosecond laser processing. By filling epoxy and UV-curing, waveguide with plastic-core and silica-cladding was created, presenting high thermal responding coefficient of 211pm/°C.

Dual-polarization multi-band OFDM signals for next generation core networks

Dual-polarization multi-band orthogonal frequency-division multiplexing (DP-MB-OFDM) signals are optimized at data rates up to 500 Gb/s in dispersion compensation fiber (DCF)-free long-haul links. We compare different polarization crosstalk compensation techniques and explore the DP-MB-OFDM transmission distance limitations. In addition, the impact of the OFDM subcarrier number and high-level modulation format on the transmission performance are also investigated. Finally, a comparison is made between DP-MB-OFDM and the industrial solution of single-carrier DP quaternary phase-shift keying (DP-QPSK) at 1000 km.

Microfiber-based inline Mach-Zehnder interferometer for dual-parameter measurement

An approach to realizing simultaneous measurement of refractive index (RI) and temperature based on a microfiber-based dual inline Mach-Zehnder interferometer (MZI) is proposed and demonstrated. Due to different interference mechanisms, as one interference between the core mode and the lower order cladding mode in the sensing single-mode fiber and the other interference between the fundamental mode and the high-order mode in the multimode microfiber, the former interferometer achieves RI sensitivity of -23.67 nm/RIU and temperature sensitivity of 81.2 pm/oC, whereas those of the latter are 3820.23 nm/RIU, and -465.7 pm/oC, respectively. The large sensitivity differences can provide a more accurate demodulation of RI and temperature. The sensor is featured with multiparameters measurement, compact structure, high sensitivity, low cost, and easy fabrication.

Three essays on dual-class stock structure

Dual-class stock structure is characterized by the separation of voting rights and cash flow rights. The departure from a common "one share-one vote" configuration creates ideal conditions for conflicts of interest and agency problems between controlling insiders (the holders of voting rights) and remaining shareholders. The owners of voting rights have the opportunity to extract private benefits and act in their personal interest; as a result, dual-class firms are often perceived to have low transparency and high information asymmetry. This dissertation investigates the quality of information and the information environment of firms with two classes of stock. The first essay examines the quality of information by studying accruals in dual-class firms in comparison to firms with only one class of stock. The results suggest that the quality of accruals is better in dual-class firms than in single-class firms. In addition, the difference in the quality of accruals between firms that abolish their dual-class share structure by unification and singe-class firms disappears in the post-unification period. The second essay investigates the earnings informativeness of dual-class firms by examining the explanatory power of earnings for returns. The results indicate that the earnings informativeness is lower for dual-class firms as compared to single-class firms. Earnings informativeness improves in firms that unify their shares. The third essay compares the level of information asymmetry between dual-class firms and single-class firms. It is documented that the information environment for dual-class firms is worse than for single-class firms. Also, the finding suggests that the difference in information environment between dual-class firms and single-class firms disappears after dual-class stock unification.

CONSTRAINTS ON THE LITHOSPHERIC STRUCTURE OF MID OCEAN RIDGES FROM OCEANIC CORE COMPLEX MORPHOLOGY

The Mid-oceanic ridge system is a feature unique to Earth. It is one of the fundamental components of plate tectonics and reflects interior processes of mantle convection within the Earth. The thermal structure beneath the mid-ocean ridges has been the subject of several modeling studies. It is expected that the elastic thickness of the lithosphere is larger near the transform faults that bound mid-ocean ridge segments. Oceanic core complexes (OCCs), which are generally thought to result from long-lived fault slip and elastic flexure, have a shape that is sensitive to elastic thickness. By modeling the shape of OCCs emplaced along a ridge segment, it is possible to constraint elastic thickness and therefore the thermal structure of the plate and how it varies along-axis. This thesis builds upon previous studies that utilize thin plate flexure to reproduce the shape of OCCs. I compare OCC shape to a suite of models in which elastic thickness, fault dip, fault heave, crustal thickness, and axial infill are systematically varied. Using a grid search, I constrain the parameters that best reproduce the bathymetry and/or the slope of ten candidate OCCs identified along the 12°—15°N segment of the Mid-Atlantic Ridge. The lithospheric elastic thicknesses that explains these OCCs is thinner than previous investigators suggested and the fault planes dip more shallowly in the subsurface, although at an angle compatible with Anderson’s theory of faulting. No relationships between model parameters and an oceanic core complexes location within a segment are identified with the exception that the OCCs located less than 20km from a transform fault have slightly larger elastic thickness than OCCs in the middle of the ridge segment.

Three Essays on Dual-Class Stock Structure

Dual-class stock structure is characterized by the separation of voting rights and cash flow rights. The departure from a common “one share-one vote” configuration creates ideal conditions for conflicts of interest and agency problems between controlling insiders (the holders of voting rights) and remaining shareholders. The owners of voting rights have the opportunity to extract private benefits and act in their personal interest; as a result, dual-class firms are often perceived to have low transparency and high information asymmetry. This dissertation investigates the quality of information and the information environment of firms with two classes of stock. The first essay examines the quality of information by studying accruals in dual-class firms in comparison to firms with only one class of stock. The results suggest that the quality of accruals is better in dual-class firms than in single-class firms. In addition, the difference in the quality of accruals between firms that abolish their dual-class share structure by unification and singe-class firms disappears in the post-unification period. The second essay investigates the earnings informativeness of dual-class firms by examining the explanatory power of earnings for returns. The results indicate that the earnings informativeness is lower for dual-class firms as compared to single-class firms. Earnings informativeness improves in firms that unify their shares. The third essay compares the level of information asymmetry between dual-class firms and single-class firms. It is documented that the information environment for dual-class firms is worse than for single-class firms. Also, the finding suggests that the difference in information environment between dual-class firms and single-class firms disappears after dual-class stock unification.

Ssr-based Genetic Diversity And Structure Of Garlic Accessions From Brazil.

Garlic is a spice and a medicinal plant; hence, there is an increasing interest in 'developing' new varieties with different culinary properties or with high content of nutraceutical compounds. Phenotypic traits and dominant molecular markers are predominantly used to evaluate the genetic diversity of garlic clones. However, 24 SSR markers (codominant) specific for garlic are available in the literature, fostering germplasm researches. In this study, we genotyped 130 garlic accessions from Brazil and abroad using 17 polymorphic SSR markers to assess the genetic diversity and structure. This is the first attempt to evaluate a large set of accessions maintained by Brazilian institutions. A high level of redundancy was detected in the collection (50 % of the accessions represented eight haplotypes). However, non-redundant accessions presented high genetic diversity. We detected on average five alleles per locus, Shannon index of 1.2, HO of 0.5, and HE of 0.6. A core collection was set with 17 accessions, covering 100 % of the alleles with minimum redundancy. Overall FST and D values indicate a strong genetic structure within accessions. Two major groups identified by both model-based (Bayesian approach) and hierarchical clustering (UPGMA dendrogram) techniques were coherent with the classification of accessions according to maturity time (growth cycle): early-late and midseason accessions. Assessing genetic diversity and structure of garlic collections is the first step towards an efficient management and conservation of accessions in genebanks, as well as to advance future genetic studies and improvement of garlic worldwide.

Morphological analysis of glass, carbon and glass/carbon fiber posts and bonding to self or dual-cured resin luting agents

OBJECTIVE: The aim of this study was to evaluate the morphology of glass (GF), carbon (CF) and glass/carbon (G/CF) fiber posts and their bond strength to self or dual-cured resin luting agents. MATERIAL AND METHODS: Morphological analysis of each post type was conducted under scanning electron microscopy (SEM). Bond strength was evaluated by microtensile test after bisecting the posts and re-bonding the two halves with the luting agents. Data were subjected to two-way ANOVA and Tukey's test (α=0.05). Failure modes were evaluated under optical microscopy and SEM. RESULTS: GF presented wider fibers and higher amount of matrix than CF, and G/CF presented carbon fibers surrounded by glass fibers, and both involved by matrix. For CF and GF, the dual-cured material presented significantly higher (p<0.05) bond strength than the self-cured agent. For the dual agent, CF presented similar bond strength to GF (p>0.05), but higher than that of G/CF (p<0.05). For the self-cured agent, no significant differences (p>0.05) were detected, irrespective of the post type. For GF and G/CF, all failures were considered mixed, while a predominance of adhesive failures was detected for CF. CONCLUSION: The bonding between fiber posts and luting agents was affected by the type of fibers and polymerization mode of the cement. When no surface treatment of the post is performed, the bonding between glass fiber post and dual-cured agent seems to be more reliable.

Defects in vesicle core induced by Escherichia coli dihydroorotate dehydrogenase

Dihydroorotate dehydrogenase (DHODH) catalyzes the oxidation of dihydroorotate to orotate during the fourth step of the de novo pyrimidine synthesis pathway. In rapidly proliferating mammalian cells, pyrimidine salvage pathway is insufficient to overcome deficiencies in that pathway for nucleotide synthesis. Moreover, as certain parasites lack salvage enzymes, relying solely on the de novo pathway, DHODH inhibition has turned out as an efficient way to block pyrimidine biosynthesis. Escherichia coli DHODH (EcDHODH) is a class 2 DHODH, found associated to cytosolic membranes through an N-terminal extension. We used electronic spin resonance (ESR) to study the interaction of EcDHODH with vesicles of 1,2-dioleoyl-sn-glycero-phosphatidylcholine/detergent. Changes in vesicle dynamic structure induced by the enzyme were monitored via spin labels located at different positions of phospholipid derivatives. Two-component ESR spectra are obtained for labels 5- and 1 0-phosphatidylcholine in presence of EcDHODH, whereas other probes show a single-component spectrum. The appearance of an additional spectral component with features related to fast-motion regime of the probe is attributed to the formation of a defect-like structure in the membrane hydrophobic region. This is probably the mechanism used by the protein to capture quinones used as electron acceptors during catalysis. The use of specific spectral simulation routines allows us to characterize the ESR spectra in terms of changes in polarity and mobility around the spin-labeled phospholipids. We believe this is the first report of direct evidences concerning the binding of class 2 DHODH to membrane systems.

Optical absorption and electronic band structure first-principles calculations of alpha-glycine crystals

Light absorption of alpha-glycine crystals grown by slow evaporation at room temperature was measured, indicating a 5.11 +/- 0.02 eV energy band gap. Structural, electronic, and optical absorption properties of alpha-glycine crystals were obtained by first-principles quantum mechanical calculations using density functional theory within the generalized gradient approximation in order to understand this result. To take into account the contribution of core electrons, ultrasoft and norm-conserving pseudopotentials, as well as an all electron approach were considered to compute the electronic density of states and band structure of alpha-glycine crystals. They exhibit three indirect energy band gaps and one direct Gamma-Gamma energy gap around 4.95 eV. The optical absorption related to transitions between the top of the valence band and the bottom of the conduction band involves O 2p valence states and C, O 2p conduction states, with the carboxyl group contributing significantly to the origin of the energy band gap. The calculated optical absorption is highly dependent on the polarization of the incident radiation due to the spatial arrangement of the dipolar glycine molecules; in the case of a polycrystalline sample, the first-principles calculated optical absorption is in good agreement with the measurement when a rigid energy shift is applied.

Electronic structure of In(2)O(3) and Sn-doped In(2)O(3) by hard x-ray photoemission spectroscopy

The valence and core levels of In(2)O(3) and Sn-doped In(2)O(3) have been studied by hard x-ray photoemission spectroscopy (hv = 6000 eV) and by conventional Al K alpha (hv = 1486.6 eV) x-ray photoemission spectroscopy. The experimental spectra are compared with density-functional theory calculations. It is shown that structure deriving from electronic levels with significant In or Sn 5s character is selectively enhanced under 6000 eV excitation. This allows us to infer that conduction band states in Sn-doped samples and states at the bottom of the valence band both contain a pronounced In 5s contribution. The In 3d core line measured at hv = 1486.6 eV for both undoped and Sn-doped In(2)O(3) display an asymmetric lineshape, and may be fitted with two components associated with screened and unscreened final states. The In 3d core line spectra excited at hv = 6000 eV for the Sn-doped samples display pronounced shoulders and demand a fit with two components. The In 3d core line spectrum for the undoped sample can also be fitted with two components, although the relative intensity of the component associated with the screened final state is low, compared to excitation at 1486.6 eV. These results are consistent with a high concentration of carriers confined close to the surface of nominally undoped In(2)O(3). This conclusion is in accord with the fact that a conduction band feature observed for undoped In(2)O(3) in Al K alpha x-ray photoemission is much weaker than expected in hard x-ray photoemission.

Crystal structure of human T cell leukemia virus type 1 gp21 ectodomain crystallized as a maltose-binding protein chimera reveals structural evolution of retroviral transmembrane proteins

Retroviral entry into cells depends on envelope glycoproteins, whereby receptor binding to the surface-exposed subunit triggers membrane fusion by the transmembrane protein (TM) subunit. We determined the crystal structure at 2.5-Angstrom resolution of the ectodomain of gp21, the TM from human T cell leukemia virus type 1. The gp21 fragment was crystallized as a maltose-binding protein chimera, and the maltose-binding protein domain was used to solve the initial phases by the method of molecular replacement. The structure of gp21 comprises an N-terminal trimeric coiled coil, an adjacent disulfide-bonded loop that stabilizes a chain reversal, and a C-terminal sequence structurally distinct from HIV type 1/simian immunodeficiency virus gp41 that packs against the coil in an extended antiparallel fashion. Comparison of the gp21 structure with the structures of other retroviral TMs contrasts the conserved nature of the coiled coil-forming region and adjacent disulfide-bonded loop with the variable nature of the C-terminal ectodomain segment. The structure points to these features having evolved to enable the dual roles of retroviral TMs: conserved fusion function and an ability to anchor diverse surface-exposed subunit structures to the virion envelope and infected cell surface. The structure of gp21 implies that the N-terminal fusion peptide is in close proximity to the C-terminal transmembrane domain and likely represents a postfusion conformation.