Effects of parylene-C photooxidation on serum-assisted glial and neuronal patterning

The increasing use of patterned neural networks in multielectrode arrays and similar devices drives the constant development and evaluation of new biomaterials. Recently, we presented a promising technique to guide neurons and glia reliably and effectively. Parylene-C, a common hydrophobic polymer, was photolithographically patterned on silicon oxide (SiO2) and subsequently activated via immersion in serum. In this article, we explore the effects of ultraviolet (UV)-induced oxidation on parylene's ability to pattern neurons and glia. We exposed parylene-C stripe patterns to increasing levels of UV radiation and found a dose-dependent reduction in the total mass of patterned cells, as well as a gradual loss of glial and neuronal conformity to the patterns. In contrast, nonirradiated patterns had superior patterning results and increased presence of cells. The reduced cell adhesion and patterning after the formation of aldehyde and carboxyl groups on UV-radiated parylene-C supports our hypothesis that cell adhesion and growth on parylene is facilitated by hydrophobic adsorption of serum proteins. We conclude that unlike other cell patterning schemes, our technique does not rely on photooxidation of the polymer. Nonetheless, the precise control of oxygenated groups on parylene could pave the way for the differential binding of proteins and other molecules on the surface, aiding in the adhesion of alternative cell types. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010

Tropical climates at the Last Glacial Maximum: a new synthesis of terrestrial palaeoclimate data. I. Vegetation, lake-levels and geochemistry.

Palaeodata in synthesis form are needed as benchmarks for the Palaeoclimate Modelling Intercomparison Project (PMIP). Advances since the last synthesis of terrestrial palaeodata from the last glacial maximum (LGM) call for a new evaluation, especially of data from the tropics. Here pollen, plant-macrofossil, lake-level, noble gas (from groundwater) and δ18O (from speleothems) data are compiled for 18±2 ka (14C), 32 °N–33 °S. The reliability of the data was evaluated using explicit criteria and some types of data were re-analysed using consistent methods in order to derive a set of mutually consistent palaeoclimate estimates of mean temperature of the coldest month (MTCO), mean annual temperature (MAT), plant available moisture (PAM) and runoff (P-E). Cold-month temperature (MAT) anomalies from plant data range from −1 to −2 K near sea level in Indonesia and the S Pacific, through −6 to −8 K at many high-elevation sites to −8 to −15 K in S China and the SE USA. MAT anomalies from groundwater or speleothems seem more uniform (−4 to −6 K), but the data are as yet sparse; a clear divergence between MAT and cold-month estimates from the same region is seen only in the SE USA, where cold-air advection is expected to have enhanced cooling in winter. Regression of all cold-month anomalies against site elevation yielded an estimated average cooling of −2.5 to −3 K at modern sea level, increasing to ≈−6 K by 3000 m. However, Neotropical sites showed larger than the average sea-level cooling (−5 to −6 K) and a non-significant elevation effect, whereas W and S Pacific sites showed much less sea-level cooling (−1 K) and a stronger elevation effect. These findings support the inference that tropical sea-surface temperatures (SSTs) were lower than the CLIMAP estimates, but they limit the plausible average tropical sea-surface cooling, and they support the existence of CLIMAP-like geographic patterns in SST anomalies. Trends of PAM and lake levels indicate wet LGM conditions in the W USA, and at the highest elevations, with generally dry conditions elsewhere. These results suggest a colder-than-present ocean surface producing a weaker hydrological cycle, more arid continents, and arguably steeper-than-present terrestrial lapse rates. Such linkages are supported by recent observations on freezing-level height and tropical SSTs; moreover, simulations of “greenhouse” and LGM climates point to several possible feedback processes by which low-level temperature anomalies might be amplified aloft.

Postglacial vegetation, fire and climate dynamics at Central Chilean Patagonia (Lake Shaman, 44ºS)

A 19 cal ka BP pollen and charcoal record from Lake Shaman (44°S; 71°W, Chile) was analyzed to establish vegetation, fire and climate dynamics of the forest-steppe ecotone in Central Chilean Patagonia. Lake Shaman record indicates that the upper Río Cisnes valley was free of ice at around 19 cal ka BP. From this date and until 14.8 cal ka BP, a grass steppe with high proportions of shrubs associated to colder and drier conditions than present developed in this area. A continuous increase of Nothofagus accompanied by a decline in the steppe shrubs and sudden dominance of paludal over aquatic plants from 11 cal ka BP was associated to effective moisture increase but still under modern values. The replacement of the cold-dry grass-shrub steppe by a similar-than-present forest-steppe ecotone suggests an increase in temperature indicating the onset of the Holocene. At the same time, moderate fire activity suggested by the charcoal record could be related to major fuel availability as consequence of Nothofagus forest expansion. Between 8 and 3 cal ka BP, the record indicates the easternmost position of the forest-steppe ecotone suggesting the highest effective moisture with the establishment of seasonality between 5 and 3 cal ka BP. From 3 cal ka BP, the record indicates a retraction of the forest-steppe ecotone accompanied by a high pollen record variability and an increased fire activity. These late changes suggest decreased effective moisture associated with a high climatic variability. At regional and extra-regional scale, climatic changes at Lake Shaman's record are mostly associated to changes (latitudinal shifts and/or strengthening/weakening) of past Southern Westerlies that were previously recorded along Patagonia from the Lateglacial to the mid-Holocene. During the Late Holocene, a regional pattern characterized by high record variability emerges throughout Central Chilean Patagonia. This variability would be related to (1) low magnitude Southern Westerlies changes probably associated to ENSO and/or SAM or (2) the complex relationships between vegetation, fire and human occupations during the last 3 cal ka.

Holocene changes in atmospheric circulation patterns as shown by lake status changes in northern Europe

Changes in lake status, a measure of relative water depth or lake level, have been reconstructed from geological and biological evidence for 87 sites in northern Europe. During the early Holocene. the lakes show conditions similar to or drier than present in a broad band across southern Britain, southern Scandinavia and into the eastern Baltic and wetter conditions along the west coast and in central Europe. This pattern is consistent with the effects of a glacial anticyclone over the Scandinavian Ice Sheet, namely enhanced southwesterly flow along the west coast and strengthened easterlies south of the ice. After c, 8000 BP a different lake status pattern was established. with conditions drier than present over much of northern Europe. Lakes higher than today were confined to the far north, the west coast, eastern Finland and western Russia. This pattern gradually attenuated after 4000 BP. Differences in lake status during the mid- to late Holocene are consistent with a strengthening of the blocking anticyclone over the Baltic Sea in summer. resulting in more meridional circulation than today. This strengthening of the blocking anticyclone during the mid-Holocene is interpreted as a consequence of insolation changes, enhanced by the fact that the Baltic Sea was larger than present.

Datasets related to in-land water for limnology and remote sensing applications: distance-to-land, distance-to-water, water-body identifier and lake-centre co-ordinates

Datasets containing information to locate and identify water bodies have been generated from data locating static-water-bodies with resolution of about 300 m (1/360 deg) recently released by the Land Cover Climate Change Initiative (LC CCI) of the European Space Agency. The LC CCI water-bodies dataset has been obtained from multi-temporal metrics based on time series of the backscattered intensity recorded by ASAR on Envisat between 2005 and 2010. The new derived datasets provide coherently: distance to land, distance to water, water-body identifiers and lake-centre locations. The water-body identifier dataset locates the water bodies assigning the identifiers of the Global Lakes and Wetlands Database (GLWD), and lake centres are defined for in-land waters for which GLWD IDs were determined. The new datasets therefore link recent lake/reservoir/wetlands extent to the GLWD, together with a set of coordinates which locates unambiguously the water bodies in the database. Information on distance-to-land for each water cell and the distance-to-water for each land cell has many potential applications in remote sensing, where the applicability of geophysical retrieval algorithms may be affected by the presence of water or land within a satellite field of view (image pixel). During the generation and validation of the datasets some limitations of the GLWD database and of the LC CCI water-bodies mask have been found. Some examples of the inaccuracies/limitations are presented and discussed. Temporal change in water-body extent is common. Future versions of the LC CCI dataset are planned to represent temporal variation, and this will permit these derived datasets to be updated.

Glacial lake drainage in Patagonia (13-8 kyr) and response of the adjacent Pacific Ocean

Large freshwater lakes formed in North America and Europe during deglaciation following the Last Glacial Maximum. Rapid drainage of these lakes into the Oceans resulted in abrupt perturbations in climate, including the Younger Dryas and 8.2 kyr cooling events. In the mid-latitudes of the Southern Hemisphere major glacial lakes also formed and drained during deglaciation but little is known about the magnitude, organization and timing of these drainage events and their e ect on regional climate. We use 16 new single-grain optically stimulated luminescence (OSL) dates to de ne three stages of rapid glacial lake drainage in the Lago General Carrera/Lago Buenos Aires and Lago Cohrane/ Pueyrredón basins of Patagonia and provide the rst assessment of the e ects of lake drainage on the Paci c Ocean. Lake drainage occurred between 13 and 8 kyr ago and was initially gradual eastward into the Atlantic, then subsequently reorganized westward into the Paci c as new drainage routes opened up during Patagonian Ice Sheet deglaciation. Coupled ocean-atmosphere model experiments using HadCM3 with an imposed freshwater surface “hosing” to simulate glacial lake drainage suggest that a negative salinity anomaly was advected south around Cape Horn, resulting in brief but signi cant impacts on coastal ocean vertical mixing and regional climate.

Diel surface temperature range scales with lake size

Ecological and biogeochemical processes in lakes are strongly dependent upon water temperature. Long-term surface warming of many lakes is unequivocal, but little is known about the comparative magnitude of temperature variation at diel timescales, due to a lack of appropriately resolved data. Here we quantify the pattern and magnitude of diel temperature variability of surface waters using high-frequency data from 100 lakes. We show that the near-surface diel temperature range can be substantial in summer relative to long-term change and, for lakes smaller than 3 km2, increases sharply and predictably with decreasing lake area. Most small lakes included in this study experience average summer diel ranges in their near-surface temperatures of between 4 and 7°C. Large diel temperature fluctuations in the majority of lakes undoubtedly influence their structure, function and role in biogeochemical cycles, but the full implications remain largely unexplored.

The Mobility of Chondroitin Sulfate in Articular and Artificial Cartilage Characterized by (13)C Magic-Angle Spinning NMR Spectroscopy

We have studied the molecular dynamics of one of the major macromolecules in articular cartilage, chondroitin sulfate. Applying (13)C high-resolution magic-angle spinning NMR techniques, the NMR signals of all rigid macromolecules in cartilage can be suppressed, allowing the exclusive detection of the highly mobile chondroitin sulfate. The technique is also used to detect the chondroitin sulfate in artificial tissue-engineered cartilage. The tissue-engineered material that is based on matrix producing chondrocytes cultured in a collagen gel should provide properties as close as possible to those of the natural cartilage. Nuclear relaxation times of the chondroitin sulfate were determined for both tissues. Although T(1) relaxation times are rather similar, the T(2) relaxation in tissue-engineered cartilage is significantly shorter. This suggests that the motions of chondroitin sulfate in data:rat and artificial cartilage different. The nuclear relaxation times of chondroitin sulfate in natural and tissue-engineered cartilage were modeled using a broad distribution function for the motional correlation times. Although the description of the microscopic molecular dynamics of the chondroitin sulfate in natural and artificial cartilage required the identical broad distribution functions for the correlation times of motion, significant differences in the correlation times of motion that are extracted from the model indicate that the artificial tissue does not fully meet the standards of the natural ideal. This could also be confirmed by macroscopic biomechanical elasticity measurements. Nevertheless, these results suggest that NMR is a useful tool for the investigation of the quality of artificially engineered tissue. (C) 2010 Wiley Periodicals, Inc. Biopolymers 93: 520-532, 2010.

Mg(2+) Ions Bind at the C-Terminal Region of Skeletal Muscle alpha-Tropomyosin

Tropomyosin (Tm) is a dimeric coiled-coil protein that polymerizes through head-to-tail interactions. These polymers bind along actin filaments and play an important role in the regulation of muscle contraction. Analysis of its primary structure shows that Tm is rich in acidic residues, which are clustered along the molecule and may from sites for divalent cation binding. In a previous study, we showed that the Mg(2+)-induced increase in stability of the C-terminal half of Tin is sensitive to imitations near the C-terminus. In the present report, we study the interaction between Mg(2+) and full-length Tin and smaller fragments corresponding to the last 65 and 26 Tin residues. Although the smaller Tin peptide (Tm(259-284(W269))) is flexible and to large extent unstructured, the larger Tm(220-284(W269)) fragments forms a coiled coil in solution whose stability increases significantly in the presence of Mg(2+). NMR analysis shows thin Mg(2+) induces chemical shift perturbations in both Tm(220-284(W269)) and Tm(259-284(W269)) in the vicinity of His276, in which are located several negatively charged residues. (C) 2009 Wiley Periodicals, Inc. Biopolymers 91: 583-590, 2009.

Acanthoscurrin Fragment 101-132: Total Synthesis at 60 degrees C of a Novel Difficult Sequence

Glycine-rich proteins (GRP), serve a variety of biological functions. Acanthoscurrin is an antimicrobial GRP isolated front hemocytes-of the Brazilian spider Acanthoscurria gomesiana. Aiming to contribute to the knowledge of the secondary structure and stepwise solid-phase synthesis of GRPs` glycine-rich domains, we attempted to prepare G(101)GGLGGGRGGGYG(113) GGGGYGGGYG(123)GGy(126)GGGKYK(132)-NH(2), acanthoscurrin C-terminal amidated fragment. Although a theoretical prediction did not indicate high aggregation potential for this peptide, repetitive incomplete aminoacylations were observed after incorporating Tyr(126) to the growing peptide-MBHA resin (Boc chemistry) at 60 degrees C. The problem was not solved by varying the coupling reagents or solvents, adding chaotropic salts to the reaction media or changing the resin/chemistry (Rink amide resin/Fmoc chemistry). Some improvement was mode when CLEAR amide resin (Fmoc chemistry) was 32 used, as it allowed for obtaining fragment (G(113)-K(132) NIR-FT-Raman spectra collected for samples of the growing peptide-MBHA, -Rink amide resin and -CLEAR amide resin revealed the presence of beta-sheet structures. Only the combination of CLEAR-amide resin, 60 degrees C, Fmoc-(Fmoc-Hmb)Gly-OH and LiCl (the last two used alternately) was able to inhibit the phenomenon, as proven by NIR-FT-Raman analysis of the growing peptide-resin, allowing the total synthesis of desired 132 fragment Gly(101)-K(132). In summary, this work describes a new difficult sequence, contributes to understanding stepwise solid-phase synthesis of this type of peptide and shows that, at least while protected and linked to a resin, this GRPs glycine-rich motif presents all early tendency to assume beta-sheet structures. (c) 2008 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 92: 65-75, 2009.

Type 8 Design G Starboard Side; SS Lake Oneida

Print no 69

Type 8 Design G Port Side; SS Lake Oneida

Print no 77

Type 8 Design C Port Side

print no 81; Discontinued stamp JUL 23 1918

Type 8 Design C Starboard Side

Print no 73; Discontinued stamp JUL 23 1918