Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)

Introduction: In this study, quasi-three-dimensional (3D) microwell patterns were fabricated with poly (l-lactic acid) for the development of cell-based assays, targeting voltage-gated calcium channels (VGCCs). Methods and materials: SH-SY5Y human neuroblastoma cells were interfaced with the microwell patterns and found to grow as two dimensional (2D), 3D, and near two dimensional (N2D), categorized on the basis of the cells’ location in the pattern. The capability of the microwell patterns to support 3D cell growth was evaluated in terms of the percentage of the cells in each growth category. Cell spreading was analyzed in terms of projection areas under light microscopy. SH-SY5Y cells’ VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium GreenTM-1. The expression of L-type calcium channels was evaluated using immunofluorescence staining with DM-BODIPY. Results: It was found that cells within the microwells, either N2D or 3D, showed more rounded shapes and less projection areas than 2D cells on flat poly (l-lactic acid) substrates. Also, cells in microwells showed a significantly lower VGCC responsiveness than cells on flat substrates, in terms of both response magnitudes and percentages of responsive cells, upon depolarization with 50 mM K+. This lower VGCC responsiveness could not be explained by the difference in L-type calcium channel expression. For the two patterns addressed in this study, N2D cells consistently exhibited an intermediate value of either projection areas or VGCC responsiveness between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness. Conclusion: These results suggest that the pattern structure and therefore the cell growth characteristics were critical factors in determining cell VGCC responsiveness and thus provide an approach for engineering cell functionality in cell-based assay systems and tissue engineering scaffolds.

(Table 3) Sulfur and oxygen isotopic composition of cold water soluble sulfate, and strontium content and isotopic composition of upper Miocene evaporite samples recovered from DSDP Site 23-227

The origin of three Red Sea submarine brine pools was investigated by analysis of the S and O isotope ratios of dissolved sulfate and Sr isotope ratios of dissolved Sr in the brines. Sulfur and O isotope ratios of sulfate and Sr isotope ratios of evaporitic source rocks for the brines were measured for comparison. The S, O and Sr isotope ratios of evaporites recovered from DSDP site 227 are consistent with an upper Miocene evaporites age. The Valdivia Deep brine formed by karstic dissolution of Miocene evaporites by overlying seawater and shows no signs of hydrothermal input. The Suakin Deep brines are derived from, or have isotopically exchanged with Miocene or older evaporites. There has been only minor dilution of the brine by overlying seawater. Strontium isotope ratios of Suakin brine may indicate addition of a minor (15%) amount of volcanic Sr to the brine, but there is no evidence of high temperature brine-rock interaction. The sulfate in the Atlantis II brine was apparently derived from seawater. The O isotope ratio of sulfate in the present Atlantis II brine could reflect isotopic exchange between seawater sulfate and the brine at approximately 255°C. Approximately 30% of the Sr in the Atlantis II brine is derived from the underlying basalt, probably by hydrothermal leaching. Atlantis II brine is the only known example from the Red Sea which has a significant high-temperature hydrothermal history.

Mapeamento das projeções retinianas para o sistema óptico acessório, colículo superior e complexo pré-tectal do sagui (callithrix jacchus)

The accessory optical system, the pretectal complex, and superior colliculus are important control centers in a variety of eye movement, being extremely necessary for image formation, consequently to visual perception. The accessory optical system is constituted by the nuclei: dorsal terminal nucleus, lateral terminal nucleus, medial terminal nucleus and interstitial nucleus of the posterior superior fasciculus. From a functional point of view they contribute to the image stabilization, participating in the visuomotor activity where all system cells respond to slow eye movements and visual stimuli, which is important for the proper functioning of other visual systems. The pretectal complex comprises a group of nuclei situated in mesodiencephalic transition, they are: anterior pretectal nucleus, posterior pretectal nucleus, medial pretectal nucleus, olivary pretectal nucleus and the nucleus of the optic tract, all retinal projection recipients and functionally are related to the route of the pupillary light reflex and the optokinetic nystagmus. The superior colliculus is an important subcortical visual station formed by layers and has an important functional role in the control of eye movements and head in response to multisensory stimuli. Our aim was to make a mapping of retinal projections that focus on accessory optical system, the nuclei of pretectal complex and the superior colliculus, searching mainly for pretectal complex, better delineation of these structures through the anterograde tracing with the B subunit of cholera toxin (CTb) followed by immunohistochemistry and characterized (measured diameter) synaptic buttons present on the fibers / terminals of the nucleus complex pré-tectal. In our results accessory optical system, including a region which appears to be medial terminal nucleus and superior colliculus, were strongly marked by fibers / terminals immunoreactive CTb as well as pretectal complex in the nucleus: optic tract, olivary pretectal nucleus, anterior pretectal nucleus and posterior pretectal nucleus. According to the characterization of the buttons it was possible to make a better definition of these nucleus.

Semi-engineered earthquake-resistant structures: one-storey buildings built up with gabion-box walls

This thesis studies the static and seismic behavior of simple structures made with gabion box walls. The analysis was performed considering a one-story building with standard dimensions in plan (6m x 5m) and a lightweight timber roof. The main focus of the present investigation is to find the principals aspects of the seismic behavior of a one story building made with gabion box walls, in order to prevent a failure due to seismic actions and in this way help to reduce the seismic risk of developing countries where this natural disaster have a significant intensity. Regarding the gabion box wall, it has been performed some calculations and analysis in order to understand the static and dynamic behavior. From the static point of view, it has been performed a verification of the normal stress computing the normal stress that arrives at the base of the gabion wall and the corresponding capacity of the ground. Moreover, regarding the seismic analysis, it has been studied the in-plane and out-of-plane behavior. The most critical aspect was discovered to be the out-of-plane behavior, for which have been developed models considering the “rigid- no tension model” for masonry, finding a kinematically admissible multiplier that will create a collapse mechanism for the structure. Furthermore, it has been performed a FEM and DEM models to find the maximum displacement at the center of the wall, maximum tension stresses needed for calculating the steel connectors for joining consecutive gabions and the dimensions (length of the wall and distance between orthogonal walls or buttresses) of a geometrical configuration for the standard modulus of the structure, in order to ensure an adequate safety margin for earthquakes with a PGA around 0.4-0.5g. Using the results obtained before, it has been created some rules of thumb, that have to be satisfy in order to ensure a good behavior of these structure.

Physical oceanography from 120 CTDs, 79 water bottles stations, and 30 helicopter-borne CTDs during POLARSTERN cruise ANT-XXII/2 (ISPOL)

The Ice Station POLarstern (ISPOL) cruise revisited the western Weddell Sea in late 2004 and obtained a comprehensive set of conductivity-temperature-depth (CTD) data. This study describes the thermohaline structure and diapycnal mixing environment observed in 2004 and compares them with conditions observed more than a decade earlier. Hydrographic conditions on the central western Weddell Sea continental slope, off Larsen C Ice Shelf, in late winter/early spring of 2004/2005 can be described as a well-stratified environment with upper layers evidencing relict structures from intense winter near-surface vertical fluxes, an intermediate depth temperature maximum, and a cold near-bottom layer marked by patchy property distributions. A well-developed surface mixed layer, isolated from the underlying Warm Deep Water (WDW) by a pronounced pycnocline and characterized by lack of warming and by minimal sea-ice basal melting, supports the assumption that upper ocean winter conditions persisted during most of the ISPOL experiment. Much of the western Weddell Sea water column has remained essentially unchanged since 1992; however, significant differences were observed in two of the regional water masses. The first, Modified Weddell Deep Water (MWDW), comprises the permanent pycnocline and was less saline than a decade earlier, whereas Weddell Sea Bottom Water (WSBW) was horizontally patchier and colder. Near-bottom temperatures observed in 2004 were the coldest on record for the western Weddell Sea over the continental slope. Minimum temperatures were ~0.4 and ~0.3 °C colder than during 1992-1993, respectively. The 2004 near-bottom temperature/salinity characteristics revealed the presence of two different WSBW types, whereby a warm, fresh layer overlays a colder, saltier layer (both formed in the western Weddell Sea). The deeper layer may have formed locally as high salinity shelf water (HSSW) that flowed intermittently down the continental slope, which is consistent with the observed horizontal patchiness. The latter can be associated with the near-bottom variability found in Powell Basin with consequences for the deep water outflow from the Weddell Sea.

Data on the biogeochemical cycle of methane in the ocean

Geological, mineralogical and microbiological aspects of the methane cycle in water and sediments of different areas in the oceans are under consideration in the monograph. Original and published estimations of formation- and oxidation rates of methane with use of radioisotope and isotopic methods are given. The role of aerobic and anaerobic microbial oxidation of methane in production of organic matter and in formation of authigenic carbonates is considered. Particular attention is paid to processes of methane transformation in areas of its intensive input to the water column from deep-sea hydrothermal sources, mud volcanoes, and cold methane seeps.

Sediment rates and structures in Bay of Kiel, Baltic Sea

Density and diversity of bottom fauna population as dependent on sediment types and water depth is largely well known in Kiel Bay. This is in contrast to structures and processes of bioturbation, although generally it has a big influence on the benthic boundary layer and its processes, e.g., the metabolism of the bottom fauna, the mechanical properties, the age dating, and the large field of chemical processes. In the densely inhabited sands and muddy sands of the shallower waters with sediment thicknesses of some decimeters only, bioturbation is usually ubiquitous, and most of the structures left are monotonously of "biodeformational" character. At greater water depths, however, where a sedimentary column of several meters of Holocene is developed, the X-ray radiographs of numerous sediment cores show heterogeneous biogenic structures with regional and stratigraphical differentiation. They are described in terms of ichnofabrics and are interpreted on ethological knowledge of the related macrobenthos species. lmportant organisms creating specific traces include the bivalve Arctica (Cyprina) islandica and the polychaete worm Pectinaria koreni. These species are abundant in Kiel Bay and produce by their crawling-plowing mode of locomotion, a characteristic biogenic stratification, the "plow-sole structure". Other typical biogenic structures are tube traces, which are left by a number of different polychaetes occurring either singly, or as U-pairs mainly in mud sediments. Although sea urchins are rare to absent in Kiel Bay, layers of their characteristic traces Scolicia occur as witness of paleohydrographic events in channel sediments of the central bay. Plow-sole traces, polychaete-tube ichnofabric, Scolicia layers and alternations of laminated and bioturbated layers are considered as building blocks of a future "ichnostratigraphy" of Kiel Bay.

Identification of unusual events in multi-channel bridge monitoring data using wavelet transform and outlier analysis

Peer reviewed

Atomic layer structure of vanadium oxide nanotubes grown on nanourchin structures

We report the detailed characterization of high quality vanadium oxide (VOx) nanotubes (NTs) and highlight the zipping of adjacent vanadate layers in such NTs formed on remarkable nanourchin structures. These nanostructures consist of high-density spherical radial arrays of NTs. The results evidence vanadate NTs with unprecedented uniformity and evidences the first report of vanadate atomic layer zipping. The NTs are ∼2 μm in length with inner diameters of 20-30 nm. The tube walls comprise scrolled triplet-layers of vanadate intercalated with organic surfactant. Such high-volume structures might be useful as open-access electrolyte scaffolds for lithium insertion-based charge storage devices.

Advancing atomic nanolithography: cold atomic Cs beam exposure of alkanethiol self-assembled monolayers

We report the results of a study into the quality of functionalized surfaces for nanolithographic imaging. Self-assembled monolayer (SAM) coverage, subsequent post-etch pattern definition and minimum feature size all depend on the quality of the Au substrate used in atomic nanolithographic experiments. We find sputtered Au substrates yield much smoother surfaces and a higher density of {111} oriented grains than evaporated Au surfaces. A detailed study of the self-assembly mechanism using molecular resolution AFM and STM has shown that the monolayer is composed of domains with sizes typically of 5-25 nm, and multiple molecular domains can exist within one Au grain. Exposure of the SAM to an optically-cooled atomic Cs beam traversing a two-dimensional array of submicron material masks ans also standing wave optical masks allowed determination of the minimum average Cs dose (2 Cs atoms per SAM molecule) and the realization of < 50 nm structures. The SAM monolayer contains many non-uniformities such as pin-holes, domain boundaries and monoatomic depressions which are present in the Au surface prior to SAM adsorption. These imperfections limit the use of alkanethiols as a resist in atomic nanolithography experiments. These studies have allowed us to realize an Atom Pencil suitable for deposition of precision quantities of material at the microand nanoscale to an active surface.

Anodic behavior of InP: film growth, porous structures and current oscillations.

We review our recent work on the anodization of InP in KOH electrolytes. The anodic oxidation processes are shown to be remarkably different in different concentrations of KOH. Anodization in 2 - 5 mol dm-3 KOH electrolytes results in the formation of porous InP layers but, under similar conditions in a 1 mol dm-3 KOH, no porous structure is evident. Rather, the InP electrode is covered with a thin, compact surface film at lower potentials and, at higher potentials, a highly porous surface film is formed which cracks on drying. Anodization of electrodes in 2 - 5 mol dm-3 KOH results in the formation of porous InP under both potential sweep and constant potential conditions. The porosity is estimated at ~65%. A thin layer (~ 30 nm) close to the surface appears to be unmodified. It is observed that this dense, near-surface layer is penetrated by a low density of pores which appear to connected it to the electrolyte. Well-defined oscillations are observed when InP is anodized in both the KOH and (NH4)2S. The charge per cycle remains constant at 0.32 C cm-2 in (NH4)2S but increases linearly with potential in KOH. Although the characteristics of the oscillations in the two systems differ, both show reproducible and well-behaved values of charge per cycle.

Fabrication of sub-10 nm lamellar structures by solvent vapour annealing of block copolymers

Fabrication of nanoscale patterns through the bottom-up approach of self-assembly of phase-separated block copolymers (BCP) holds promise for nanoelectronics applications. For lithographic applications, it is useful to vary the morphology of BCPs by monitoring various parameters to make “from lab to fab” a reality. Here I report on the solvent annealing studies of lamellae forming polystyrene-blockpoly( 4-vinylpyridine) (PS-b-P4VP). The high Flory-Huggins parameter (χ = 0.34) of PS-b-P4VP makes it an ideal BCP system for self-assembly and template fabrication in comparison to other BCPs. Different molecular weights of symmetric PS-b-P4VP BCPs forming lamellae patterns were used to produce nanostructured thin films by spin-coating from mixture of toluene and tetrahydrofuran(THF). In particular, the morphology change from micellar structures to well-defined microphase separated arrangements is observed. Solvent annealing provides a better alternative to thermal treatment which often requires long annealing periods. The choice of solvent (single and dual solvent exposure) and the solvent annealing conditions have significant effects on the morphology of films and it was found that a block neutral solvent was required to realize vertically aligned PS and P4VP lamellae. Here, we have followed the formation of microdomain structures with time development at different temperatures by atomic force microscopy (AFM). The highly mobilized chains phase separate quickly due to high Flory-Huggins (χ) parameter. Ultra-small feature size (~10 nm pitch size) nanopatterns were fabricated by using low molecular weight PSb- P4VP (PS and P4VP blocks of 3.3 and 3.1 kg mol-1 respectively). However, due to the low etch contrast between the blocks, pattern transfer of the BCP mask is very challenging. To overcome the etch contrast problem, a novel and simple in-situ hard mask technology is used to fabricate the high aspect ratio silicon nanowires. The lamellar structures formed after self-assembly of phase separated PS-b-P4VP BCPs were used to fabricate iron oxide nanowires which acted as hard mask material to facilitate the pattern transfer into silicon and forming silicon nanostructures. The semiconductor and optical industries have shown significant interest in two dimensional (2D) molybdenum disulphide (MoS2) as a potential device material due to its low band gap and high mobility. However, current methods for its synthesis are not ‘fab’ friendly and require harsh environments and processes. Here, I also report a novel method to prepare MoS2 layered structures via self-assembly of a PS-b-P4VP block copolymer system. The formation of the layered MoS2 was confirmed by XPS, Raman spectroscopy and high resolution transmission electron microscopy.

Temperatures, salinity, wind speed, grain sizes and foraminifera abundance in the Great Belt Channel, western Baltic Sea

The Great Belt, the largest inlet to the Baltic Sea, has a deep and well defined channel system. A distinct thermohaline layer at roughly 18 to 20 m of water depth separates the saltier and generally cooler deeper North Sea water from the brackish and warmer surface water. It is practically a current dominated area, with the strongest bottom currents due to prolonged west winds. The size and shape of the surface sediments and their grain size distributions show a close relationship with the prevailing hydrographical conditions. Southerly current marks predominate while northerly directions are confined to 10 to 14 m of water depth. The degree of bioturbation is highest in the uppermost sedimentary cover where practically all original stratification has been destroyed. Various bioturbate structures have been identified with the fauna. Coiling ratios of Ammonia beccarii (Linnaeus) have been successfully applied for correlation in the postglacial sediments of the early Littorina Transgression. The succession shows that in the Boreal brackish water conditions were probably followed by peat and limnic sediments as the sea regressed. With the Littorina Transgression, the sea again entered the area and high sedimentation rates resulted in the major deposits of the Great Belt. At least for the last 4000 years, sedimentation rates had been very low. Present day currents sweep out the sediments, mainly to the southern marginal areas.

RELIBILITY ANALYSIS OF SIMPLE SLOPES AND SOIL-STRUCTURES WITH LINEAR LIMIT STATES

The first objective of this research was to develop closed-form and numerical probabilistic methods of analysis that can be applied to otherwise conventional methods of unreinforced and geosynthetic reinforced slopes and walls. These probabilistic methods explicitly include random variability of soil and reinforcement, spatial variability of the soil, and cross-correlation between soil input parameters on probability of failure. The quantitative impact of simultaneously considering the influence of random and/or spatial variability in soil properties in combination with cross-correlation in soil properties is investigated for the first time in the research literature. Depending on the magnitude of these statistical descriptors, margins of safety based on conventional notions of safety may be very different from margins of safety expressed in terms of probability of failure (or reliability index). The thesis work also shows that intuitive notions of margin of safety using conventional factor of safety and probability of failure can be brought into alignment when cross-correlation between soil properties is considered in a rigorous manner. The second objective of this thesis work was to develop a general closed-form solution to compute the true probability of failure (or reliability index) of a simple linear limit state function with one load term and one resistance term expressed first in general probabilistic terms and then migrated to a LRFD format for the purpose of LRFD calibration. The formulation considers contributions to probability of failure due to model type, uncertainty in bias values, bias dependencies, uncertainty in estimates of nominal values for correlated and uncorrelated load and resistance terms, and average margin of safety expressed as the operational factor of safety (OFS). Bias is defined as the ratio of measured to predicted value. Parametric analyses were carried out to show that ignoring possible correlations between random variables can lead to conservative (safe) values of resistance factor in some cases and in other cases to non-conservative (unsafe) values. Example LRFD calibrations were carried out using different load and resistance models for the pullout internal stability limit state of steel strip and geosynthetic reinforced soil walls together with matching bias data reported in the literature.