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At least 523 records · Page 29Linked to original sources
PubMed2005

Effect of molecular binding to a semiconductor on metal/molecule/semiconductor junction behavior.

Diodes made by (indirectly) evaporating Au on a monolayer of molecules that are adsorbed chemically onto GaAs, via either disulfide or dicarboxylate groups, show roughly linear but opposite dependence of their effective barrier height on the dipole moment of the molecules. We explain this by Au-molecule (electrical) interactions not only with the exposed end groups of the molecule but also with its binding groups. We arrive at this conclusion by characterizing the interface by in situ UPS-XPS, ex situ XPS, TOF-SIMS, and Kelvin probe measurements, by scanning microscopy of the surfaces, and by current-voltage measurements of the devices. While there is a very limited interaction of Au with the dicarboxylic binding groups, there is a much stronger interaction with the disulfide groups. We suggest that these very different interactions lead to different (growth) morphologies of the evaporated gold layer, resulting in opposite effects of the molecular dipole on the junction barrier height.

Journal Article↗
PubMed2000

A scanning tunnelling study of immobilised cytochrome P450cam.

A site-specifically engineered surface cysteine residue, located in a region where the haem moiety is closest to the surface, is used to anchor cytochrome P450cam enzyme molecules covalently to a gold electrode. More reproducibly ordered adsorption, at high coverage, occurs with this K344C mutant than with the wild-type enzyme. The subsequently formed close-packed monolayer arrays have been probed by scanning tunnelling microscopy under ambient conditions and under aqueous (buffered) solution at high resolution. Initial indications suggest that the immobilised enzyme is both electrochemically addressable and catalytically active.

Camphor 5-Monooxygenase↗
PubMedDate not supplied

Modified fabrication process for aperture probe cantilevers.

We report the development of cantilever- and fibre-based probes for scanning near-field optical microscopy. Both probe concepts rely on the integration of a microfabricated aperture tip with reproducible optical and mechanical properties. Numerical calculations were carried out using a finite integration code to investigate the polarization-sensitive transmission behaviour of aperture tips. In order to establish technological guidelines for the optimization of the properties of the optical tip the distinct influence of the tip geometry on the intensity distribution in the vicinity of the aperture is studied in detail.

Journal Article↗
PubMed1991

Reactions of the subunits of the class II major histocompatibility complex molecule IAd.

Major histocompatibility complex (MHC) class II molecules are heterodimers formed by noncovalent linkage of alpha and beta chains. It has been shown that the subunits of the MHC class II molecules IAd and IEk bind antigenic peptides as well as antigenic peptides labeled with fluorescent probes. Laser scanning fluorescence microscopy on SDS/polyacrylamide gels demonstrates that the subunit-peptide complexes of IAd are stable over a wide pH range. Below pH 5.3 the heterodimer of IAd dissociates into the free chains, which still bind antigenic peptides such as the 18-amino acid peptide obtained by a tyrosine addition to a chicken ovalbumin peptide, Ova-(323-339)Y. The stability of preformed subunit complexes with fluorescein-labeled Ova-(323-339)Y was investigated by using high-performance size exclusion chromatography and epifluorescence microscopy. Each subunit forms a long-lived complex, both in detergent solutions and in reconstituted lipid bilayers. At 37 degrees C and pH 7.0 the dissociation half-time of the beta-subunit-peptide complex was determined to be 28 hr and that of the alpha-subunit-peptide complex was 10 hr. In contrast to the dissociation of the peptide from the IAd heterodimer, the half-times for dissociation of the peptide from the separate chains are not decreased at pH 5.0.

Animals↗
PubMed1999

Neuronal calcium sparks and intracellular calcium "noise".

Intracellular calcium ions are involved in many forms of cellular function. To accommodate so many control functions, a complex spatiotemporal organization of calcium signaling has developed. In both excitable and nonexcitable cells, calcium signaling was found to fluctuate. Sudden localized increases in the intracellular calcium concentration-or calcium sparks-were found in heart, striated and smooth muscle, Xenopus Laevis oocytes, and HeLa and P12 cells. In the nervous system, intracellular calcium ions were found important in key processes such as transmitter release, repetitive firing, and gene expression. Hence, we examined whether calcium sparks also exist in neurons. Using confocal laser-scanning microscopy and fluorescent probes, we found that calcium sparks exist in two types of neuronal preparations: the presynaptic boutons of the lizard neuromuscular junction and rat hippocampal neurons in cell culture. Control experiments exclude the possibility that these calcium sparks originate from instrumental or biological artifacts. Calcium sparks seem to be just the tip of the iceberg of a more general phenomenon of intracellular calcium "noise." We speculate that calcium sparks and calcium noise may be of key importance in calcium signaling in the nervous system.

Animals↗
PubMed2006

The effect of perinatal TCDD exposure on caries susceptibility in rats.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), the model compound of polychlorinated dibenzo-p-dioxins and furans, is a potent toxicant with the ability to hamper development. Accidental exposure to TCDD has been linked with various developmental dental aberrations in humans, and experimentally it has been shown that TCDD causes, among other defects, hypomineralization of dental hard tissues in rodents. Here, we studied the effect of very low perinatal TCDD exposure on dental caries susceptibility and mineral composition of tooth enamel in rats. Pregnant line C rats (rat line developed in our laboratory) were dosed 0.03-1.0 microg/kg TCDD on gestation day 15 and allowed to give birth and nurse until weaning on postnatal day 21. The offspring were challenged with cariogenic treatment including sugar-rich diet and three inoculations with Streptococcus mutans. Control groups involved animals with or without cariogenic challenge or TCDD treatment. The number of caries lesions in left lower molars was determined by Schiff's staining after 8 weeks of weaning. TCDD treatment increased cariogenic lesions in the enamel at the lowest maternal dose used, 0.03 microg/kg, and at the highest maternal dose, 1 microg/kg, the lesions extended through the enamel to dentin more frequently. Changes in mineral composition measured by electron probe microanalyzer, scanning electron microscopy, and energy-dispersive spectrometry could not explain the increased caries susceptibility. In conclusion, perinatal TCDD exposure can render rat molars more susceptible to caries.

Abnormalities, Drug-Induced↗
PubMed2002

Imaging subsurface reflection phase with quantized electrons.

Lead quantum wells (QW) epitaxially grown on annealed Pb/Si(111) interface form a model system for the study of interactions between quantized electrons and adiabatically modulated boundaries. Tunnel spectra of this system reveal a previously unknown adiabatic shift of QW resonances due to lateral variations of the electronic reflection phase at the buried interface. With this effect, lateral distribution of the subsurface reflection phase can be probed, using scanning tunneling microscopy.

Journal Article↗
PubMed2005

Plasmonics-based nanostructures for surface-enhanced Raman scattering bioanalysis.

Surface-enhanced Raman scattering (SERS) spectroscopy is a plasmonics-based spectroscopic technique that combines modern laser spectroscopy with unique optical properties of metallic nanostructures, resulting in strongly increased Raman signals when molecules are adsorbed on or near nanometer-size structures of special metals such as gold, silver, and transition metals. This chapter provides a synopsis of the development and application of SERS-active metallic nanostructures, especially for the analysis of biologically relevant compounds. Some highlights of this chapter include reports of SERS as an immunoassay readout method, SERS gene nanoprobes, near-field scanning optical microscopy SERS probes, SERS as a tool for single-molecule detection, and SERS nanoprobes for cellular studies.

DNA↗
PubMed2006

Ecophysiology and niche differentiation of Nitrospira-like bacteria, the key nitrite oxidizers in wastewater treatment plants.

Nitrite-oxidizing bacteria of the genus Nitrospira are key nitrifiers in wastewater treatment plants. Pure cultures of these organisms are unavailable, but cultivation-independent molecular methods make it possible to detect Nitrospira-like bacteria in environmental samples and to investigate their ecophysiology. Comprehensive screening of natural and engineered habitats and of public databases for 16S rRNA sequences of Nitrospira-like bacteria revealed a surprisingly high biodiversity in the genus Nitrospira, which comprises at least four phylogenetic sublineages. All Nitrospira-like bacteria detected in wastewater treatment plants belonged to the sublineages I and II. Subsequently, the population dynamics of different Nitrospira-like bacteria were monitored, by quantitative fluorescence in situ hybridization with rRNA-targeted probes, confocal laser scanning microscopy and digital image analysis, during incubation of nitrifying activated sludge in media containing different nitrite concentrations. These experiments showed that Nitrospira-like bacteria, which were affiliated with the phylogenetic sublineages I or II of the genus Nitrospira, responded differently to nitrite concentration shifts. Previously unknown properties of Nitrospira-like bacteria were discovered in the course of an environmental genomics project. Implications of the obtained results for fundamental understanding of the microbial ecology of nitrite oxidizers as well as for future improvement of nutrient removal in wastewater treatment plants are discussed.

Bacteria↗
PubMed2004

Cationic lipid-DNA complexes for gene therapy: understanding the relationship between complex structure and gene delivery pathways at the molecular level.

Cationic liposomes (CLs) are used as gene vectors (carriers) in worldwide human clinical trials of non-viral gene therapy. These lipid-gene complexes have the potential of transferring large pieces of DNA of up to 1 million base-pairs into cells. As our understanding of the mechanisms of action of CL-DNA complexes remains poor, transfection efficiencies are still low when compared to gene delivery with viral vectors. We describe recent studies with a combination of techniques (synchrotron x-ray diffraction for structure determination, laser-scanning confocal microscopy to probe the interactions of CL-DNA particles with cells, and luciferase reporter-gene expression assays to measure transfection efficiencies in mammalian cells), which collectively are beginning to unravel the relationship between the distinctly structured CL-DNA complexes and their transfection efficiency. The work described here is applicable to transfection optimization in ex vivo cell transfection, where cells are removed and returned to patients after transfection. CL-DNA complexes primarily form a multilayered sandwich structure with DNA layered between the cationic lipids (labeled L(alpha)(C)). On rare occasions, an inverted hexagonal structure with DNA encapsulated in lipid tubules (labeled H(II)(C)) is observed. A major recent insight is that for L(alpha)(C) complexes the membrane charge density sigma(M) of the CL-vector, rather than the charge of the cationic lipid alone, is a key universal parameter that governs the transfection efficiency of L(alpha)(C) complexes in cells. The parameter sigma(M) is a measure of the average charge per unit area of the membrane, thus taking into account the amount of neutral lipids. In contrast to L(alpha)(C) complexes, H(II)(C) complexes containing the lipid 1,2-dioleoyl-sn-glycerophosphatidylethanolamine (DOPE) exhibit no dependence on sigma(M). The current limiting factor to transfection by cationic lipid vectors appears to be the tight association of a fraction of the delivered exogenous DNA with cationic cellular molecules, which may prevent optimal transcriptional activity. Future directions are outlined, which make use of surface-functionalized CL-DNA complexes suitable for transfection in vivo.

Animals↗
PubMedDate not supplied

[Interaction of nonelectrolytes, the derivatives of 5-hydroxybenzimidazole, with erythrocyte membrane].

The method of spin probe and scanning electron microscopy were used to study the effects of some new synthetic antioxidants and bioregulators, the derivatives of 5-hydroxybenzimidazole, on the membrane structure and morphology of erythrocytes. Analysis of EPR spectra and electron micrographs revealed that the derivatives with various side substituents affect the membrane structure and shape of erythrocytes in a concentration-dependent manner, the effect correlating with the hydrophobic properties of the side derivatives. It was shown that all the compounds in the concentration range 1.10(-7) - 1.10(-3) M exhibit the echinocytogenic action, the most profound effect being found in the compound with benzyl- and ethoxygroup in sites 2 and 5, respectively. Our data suggest that nonelectrolytes, the derivatives of 5-hydroxybenzimidazole, are located in the outer monolayer of erythrocyte membrane.

Animals↗
PubMed2005

Lipoplex structures and their distinct cellular pathways.

Cationic liposomes (CLs) are used as non-viral vectors in worldwide clinical trials of gene therapy. Among other advantages, CL-DNA complexes have the ability to transfer very large genes into cells. However, since the understanding of their mechanisms of action is still incomplete, their transfection efficiencies remain low compared to those of viruses. We describe recent studies which have started to unravel the relationship between the distinct structures and physicochemical properties of CL-DNA complexes and their transfection efficiency by combining several techniques: synchrotron X-ray diffraction for structure determination, laser-scanning confocal microscopy to probe the interactions of CL-DNA particles with cells, and luciferase reporter-gene expression assays to measure transfection efficiencies in mammalian cells. Most CL-DNA complexes form a multilayered structure with DNA sandwiched between the cationic lipids (lamellar complexes, LalphaC). Much more rarely, an inverted hexagonal structure (HIIC) with single DNA strands encapsulated in lipid tubules is observed. An important recent insight is that the membrane charge density sigmaM of the CL-vector, rather than, for example, the charge of the cationic lipid, is a universal parameter governing the transfection efficiency of LalphaC complexes. This has led to a new model of the intracellular release of LalphaC complexes, through activated fusion with endosomal membranes. In contrast to LalphaC complexes, HIIC complexes exhibit no dependence on sigmaM, since their structure leads to a distinctly different mechanism of cell entry. Surface-functionalized complexes with poly(ethyleneglycol)-lipids (PEG-lipids), potentially suitable for transfection in vivo, have also been investigated, and the novel aspects of these complexes are discussed.

Biogenic Polyamines↗
PubMed1997

Advances in the microscopy of osteoarthritis.

This review describes recent contributions made by microscopy to the understanding of osteoarthritis, a clinical syndrome the pathological features of which are well defined by classical white light microscopy. The fluorescence and reflected light, conventional and scanning optical microscopy of excised osteoarthritic tissue preparations, from human and animal sources, has enabled the identification of cell proteins such as S100, of matrix components such as the proteoglycans and collagens, and of adhesion molecules including fibronectin, the integrins and tenascin. Comparable microscopic studies have been made of cell and tissue culture preparations of osteoarthritic cartilage and synovium. Scanning optical microscopy also allows the rapid measurement, in hydrated osteoarthritic tissues, of cell density, cell size, surface roughness and other parameters. The importance of water in sustaining the physical attributes of cartilage is accepted and new forms of electron microscopy can play important parts in the study of unfixed osteoarthritic cartilage. These methods include the low temperature scanning electron microscopy and electron probe x-ray microanalysis of hydrated bulk material and the high resolution transmission electron microscopy of low temperature replicas of cartilage surfaces. Understanding of osteoarthritis has been facilitated by these advances and will continue to be enhanced as new techniques of microscopy evolve.

Animals↗
PubMed2000

A biodegradable hybrid sponge nested with collagen microsponges.

A biodegradable hybrid sponge of poly(DL-lactic-co-glycolic acid) (PLGA) and collagen was fabricated by forming microsponges of collagen in the pores of PLGA sponge. Observation of the PLGA-collagen hybrid sponge by scanning electron microscopy (SEM) showed that microsponges of collagen with interconnected pore structures were formed in the pores of PLGA sponge. The hybrid structure further was confirmed by scanning electron microscopy-electron probe microanalysis (SEM-EPMA), and elemental nitrogen was detected in the microsponges of collagen and on the pore surfaces of PLGA, but not in cross-sections of PLGA regions. The formation of collagen microsponges was dependent on collagen concentration, the effective range of which was from 0.1 to 1.5 (w/v) %. The mechanical strength of the hybrid sponge was higher than that of either PLGA or collagen sponges, in both dry and wet states. The wettability with water was improved by hybridization with collagen, which facilitated cell seeding in the hybrid sponge. Mouse fibroblast L929 cells attached well and spread on the surfaces of the microsponges of collagen in the hybrid sponge. The distribution of cells was spatially uniform throughout the hybrid sponge. Use of the PLGA sponge as a skeleton facilitated formation of the hybrid sponge into desired shapes with high mechanical strength while collagen microsponges contributed good cell interaction and hydrophilicity.

Animals↗
PubMed1986

Early alterations at the plasma membrane of breast cancer cell lines in response to estradiol and hydroxytamoxifen.

The time course of the early stage of estradiol-17 beta (E2) and hydroxytamoxifen (OHTAM) action at the plasma membrane of hormone-responsive MCF-7 and non-responsive MDA-MB-231 (MDA) breast cancer cell lines was investigated using scanning electron microscopy (SEM), electron probe X-ray microanalysis and microelectrophysiology analysis. SEM showed a marked increase in the density and the length of microvilli (MV) on MCF-7 cells treated with 1 nM estradiol for 1 min. This membrane response disappeared at 5 min. No early effect was obtained with OHTAM, but both compounds produced a similar surge of heterogeneous MV at 15 min of treatment. The morphological change induced by E2 subsided at 60 min, whereas that of OHTAM persisted. X-ray microanalysis and computer determination of peak/background ratios permitted the demonstration that these morphological alterations were concomitant with a rise in the intracellular level of potassium. Microelectrophysiology analysis showed a sharp transitory decrease in the membrane potential of MCF-7 cells in response to estradiol. In the estrogen-insensitive MDA cells, the hormone did not modify the membrane potential and K levels decreased at 1 and 5 min before rising again to control levels at minute 15 when MV appeared. With OHTAM, potassium decreased significantly at 60 min of treatment. These initial and transitory changes in surface morphology paralleled by alterations in potassium level may be consistent with the occurrence of estrogen membrane receptors on target cells, a new aspect of steroid hormone action.

Breast Neoplasms↗
PubMed2000

Probing the building blocks of eumelanins using scanning electron microscopy.

Scanning electron microscopy (SEM) is used to examine the structure of natural and synthetic melanins. Eumelanin from Sepia officinalis and synthetic eumelanin are found to be structurally dissimilar. The natural sample has a significant structural order with subunits that have a lateral dimension of approximately 15 nm. The synthetic samples, on the other hand, appear to be amorphous solids. These results lend support for the existence of fundamental structural units proposed from the analyses of wide-angle X-ray diffraction measurements and previous mass-spectrometry results. These findings also provide insight into the disparate photophysical behavior of Sepia and synthetic eumelanin.

Animals↗
PubMed2004

HAADF-STEM imaging with sub-angstrom probes: a full Bloch wave analysis.

A full coherent Bloch wave calculation is presented to investigate high-angle annular dark-field image formation for sub-angstrom probes in scanning transmission electron microscopy (STEM). With increasing illumination angle, the contribution of the 1s bound state increases to a maximum at an optimum probe angle, after which we find increasing contributions from high-angle plane wave states around the periphery of the objective aperture. Examination of image contributions from different depths within a crystal shows an oscillatory behavior due to the beating between 1s and non-1s states. The oscillation period reduces with decreasing probe size, while the relative contribution from a specific depth increases. This signifies a changeover from a projection mode of imaging to a depth-slicing mode of imaging. This new mode appears capable of resolving three-dimensional atomic structures in future generation aberration-corrected STEM.

Algorithms↗