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Life Sciences57%

Selected Abstracts

Simulated microgravity activates MAPK pathways in fibroblasts cultured on microgrooved surface topography

CYTOSKELETON, Issue 2 2008
W. A. Loesberg
Abstract This study evaluated in vitro the differences in morphological behaviour between fibroblast cultured on smooth and microgrooved substrata (groove depth: 0.5 ,m, width: 1 ,m), which were subjected to simulated microgravity. The aim of the study was to clarify which of these parameters was more dominant to determine cell behaviour. Morphological characteristics were investigated using scanning electron microscopy and fluorescence microscopy in order to obtain qualitative information on cell alignment. Expression of collagen type I, and ,1-, ,1-, ,3-integrin were investigated by QPCR. Finally, immunoblotting was applied to visualise MAPK signalling pathways. Microscopy and image analysis showed that the fibroblasts aligned along the groove direction on all textured surfaces. On the smooth substrata, cells had spread out in a random fashion. The alignment of cells cultured on grooved surfaces under simulated microgravity, after 48 h of culturing appeared similar to those cultured at 1g, although cell shape was different. Analysis of variance proved that all main parameters: topography, gravity force, and time were significant. In addition, gene levels were reduced by simulated microgravity particularly those of ,3-integrin and collagen, however alpha-1 and beta-1 integrin levels were up-regulated. ERK1/2 was reduced in RPM, however, JNK/SAPK and p38 remained active. The members of the small GTPases family were stimulated under microgravity, particularly RhoA and Cdc42. The results are in agreement that application of microgravity to fibroblasts promotes a change in their morphological appearance and their expression of cell-substratum proteins through the MAPK intracellular signalling pathways. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source]

The effect of combined simulated microgravity and microgrooved surface topography on fibroblasts

CYTOSKELETON, Issue 3 2007
W. A. Loesberg
Abstract This study evaluated in vitro the differences in morphological behaviour between fibroblast cultured on smooth and microgrooved substrata (groove depth: 0.5 ,m, width: 1, 2, 5, and 10 ,m), which were subjected to simulated microgravity. The aim of the study was to clarify which of these parameters was more dominant to determine cell behaviour. Morphological characteristics were investigated using scanning electron microscopy and fluorescence microscopy in order to obtain qualitative information on cell alignment and area. Confocal laser scanning microscopy visualised distribution of actin filaments and focal adhesion points. Finally, expression of collagen type I, fibronectin, and ,1- and ,1-integrin were investigated by PCR. Microscopy and image analysis showed that the fibroblasts aligned along the groove direction on all textured surfaces. On the smooth substrata, cells had spread out in a random fashion. The alignment of cells cultured on grooved surfaces decreased under simulated microgravity, especially after 24 h of culturing. Cell surface area on grooved substrata were significantly smaller than on smooth substrata, but simulated microgravity on the grooved groups resulted in an enlargement of cell area. ANOVA was performed on all main parameters: topography, gravity force, and time. In this analysis, all parameters proved significant. In addition, gene levels were reduced by microgravity particularly those of ,1-integrin and fibronectin. From our data it is concluded that the fibroblasts primarily adjust their shape according to morphological environmental cues like substratum surface whilst a secondary, but significant, role is played by microgravity conditions. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source]

The effect of combined hypergravity and microgrooved surface topography on the behaviour of fibroblasts

CYTOSKELETON, Issue 7 2006
W. A. Loesberg
Abstract This study evaluated in vitro the differences in morphological behaviour between fibroblast cultured on smooth and microgrooved substrata (groove depth: 1 ,m, width: 1, 2, 5, 10 ,m), which undergo artificial hypergravity by centrifugation (10, 24 and 50 g; or 1 g control). The aim of the study was to clarify which of these parameters was more important to determine cell behaviour. Morphological characteristics were investigated using scanning electron microscopy and fluorescence microscopy in order to obtain qualitative information on cell spreading and alignment. Confocal laser scanning microscopy visualised distribution of actin filaments and vinculin anchoring points through immunostaining. Finally, expression of collagen type I, fibronectin, and ,1 - and ,1 -integrin were investigated by PCR. Microscopy and image analysis showed that the fibroblasts aligned along the groove direction on all textured surfaces. On the smooth substrata (control), cells spread out in a random fashion. The alignment of cells cultured on grooved surfaces increased with higher g-forces until a peak value at 25 g. An ANOVA was performed on the data, for all main parameters: topography, gravity force, and time. In this analysis, all parameters proved significant. In addition, most gene levels were reduced by hypergravity. Still, collagen type 1 and fibronectin are seemingly unaffected by time or force. From our data it is concluded that the fibroblasts primarily adjust their shape according to morphological environmental cues like substratum surface whilst a secondary, but significant, role is played by hypergravity forces. Cell Motil. Cytoskeleton 2006. © 2006 Wiley-Liss, Inc. [source]

Testing and improving experimental parameters for the use of low molecular weight targets in array-CGH experiments,

HUMAN MUTATION, Issue 11 2006
Marianne Stef
Abstract Array,comparative genomic hybridization (CGH) has evolved as a useful technique for the detection and characterization of deletions, and, to a lesser extent, of duplications. The resolution of the technique is dictated by the genomic distance between targets spotted on the microarray, and by the targets' sizes. The use of region-specific, high-resolution microarrays is a specific goal when studying regions that are prone to rearrangements, such as those involved in deletion syndromes. The aim of the present study was to evaluate the best experimental conditions to be used for array-CGH analysis using low molecular weight (LMW) targets. The parameters tested were: the target concentration, the way LMW targets are prepared (either as linearized plasmids or as purified PCR products), and the way the targets are attached to the array-CGH slide (in a random fashion on amino-silane coated slides, or by one amino-modified end on epoxysilane-coated slides). As a test case, we constructed a microarray harboring LMW targets located in the CREBBP gene, mutations of which cause the Rubinstein-Taybi syndrome (RTS). From 10 to 15% of RTS patients have a CREBBP deletion. We showed that aminosilane- and epoxysilane-coated slides were equally efficient with targets above 1,000,bp in size. On the other hand, with the smallest targets, especially those below 500,bp, epoxysilane-coated slides were superior to aminosilane-coated slides, which did not allow deletion detection. Use of the high resolution array allowed us to map intragenic breakpoints with precision and to identify a very small deletion and a duplication that were not detected by the currently available techniques for finding CREBBP deletions. Hum Mutat 27(11), 1143,1150, 2006. © 2006 Wiley-Liss, Inc. [source]

Random and localized resistive switching observation in Pt/NiO/Pt

PHYSICA STATUS SOLIDI - RAPID RESEARCH LETTERS, Issue 6 2007
Jung-Bin Yun
Abstract Resistive memory switching devices based on transition metal oxides are now emerging as a candidate for nonvolatile memories. To visualize nano-sized (10 nm to 30 nm in diameter) conducting filamentary paths in the surface of NiO thin films during repetitive switching, current sensing,atomic force microscopy and ultra-thin (<5 nm) Pt films as top electrodes were used. Some areas (or spots), which were assumed to be the beginning of the conducting filaments, appeared (formation) and disappeared (rupture) in a localized and random fashion during the switching and are thought to contribute to resistive memory switching. (© 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source]

Bedside Detection of Urine ,-Hydroxybutyrate in Diagnosing Metabolic Acidosis

ACADEMIC EMERGENCY MEDICINE, Issue 8 2008
Silas W. Smith MD
Abstract Objectives:, While critically important, the rapid identification of the etiology of metabolic acidosis (MA) may be labor-intensive and time-consuming. Alcoholic, starvation, and severe diabetic ketoacidosis (AKA, SKA, and DKA, respectively) may produce ,-hydroxybutyrate (BOHB) in marked excess of acetone (ACET) and acetoacetate (AcAc). Unfortunately, current urine dipstick technology poorly detects ACET and cannot measure BOHB. The inability to detect BOHB might delay therapy for ketoacidoses or provoke unnecessary evaluation or empiric treatment of other causes of MA, such as toxic alcohol poisoning. The authors tested the previous assertion that commonly available hydrogen peroxide (H2O2) would improve BOHB detection. The effectiveness of alkalinization and use of a silver nitrate (AgNO3) catalyst was also assessed. Methods:, Control and urine test specimens containing from 0.5 to 800 mmol/L ACET, AcAc, and BOHB were prepared. Urine specimens were oxidized with H2O2 (3%) 1:9 (H2O2:urine), alkalinized with potassium hydroxide (KOH; 10%), exposed to AgNO3 sticks, or altered with a combination of these methods in a random fashion. Three emergency physicians (EPs) blinded to the preparation technique evaluated urine dipsticks (Multistix, Bayer Corp.) placed in the specimens for "ketones." Results:, Multistix detected AcAc appropriately; ACET was detected only at high concentrations of ,600 mmol/L. Multistix failed to measure BOHB at all concentrations tested. H2O2 improved urinary BOHB detection, although not to clinically relevant levels (40 mmol/L). Alkalinization and AgNO3 sticks did not improve BOHB detection beyond this threshold. Conclusions:, Addition of H2O2 (3%), alkalinization, or AgNO3 sticks did not improve clinically meaningful urine BOHB detection. Clinicians should use direct methods to detect BOHB when suspected. [source]

Sketches from a Design Process: Creative Cognition Inferred From Intermediate Products

COGNITIVE SCIENCE - A MULTIDISCIPLINARY JOURNAL, Issue 1 2005
Saskia Jaarsveld
Abstract Novice designers produced a sequence of sketches while inventing a logo for a novel brand of soft drink. The sketches were scored for the presence of specific objects, their local features and global composition. Self-assessment scores for each sketch and art critics' scores for the end products were collected. It was investigated whether the design evolves in an essentially random fashion or according to an overall heuristic. The results indicated a macrostructure in the evolution of the design, characterized by two stages. For the majority of participants, the first stage is marked by the introduction and modification of novel objects and their local and global aspects; the second stage is characterized by changes in their global composition. The minority that showed the better designs has a different strategy, in which most global changes were made in the beginning. Although participants did not consciously apply these strategies, their self-assessment scores reflect the stages of the process. [source]