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PubMed2006

With a grain of salt: what halite has to offer to discussions on the origin of life.

This experimental study investigated how the dynamics of the crystallization of the evaporite mineral halite could affect the accumulation and preservation of organic macromolecules present in the crystallizing solution. Halite was grown under controlled conditions in the presence of polymer nanoparticles that acted as an analog to protocellular material. Optical microscopy, atomic force microscopy, and laser scanning confocal fluorescence microscopy were used to trace the localization of the nanoparticles during and after growth of halite crystals. The present study revealed that the organic nanoparticles were not regularly incorporated within the halite, but were very concentrated on its surfaces. Their distribution was controlled dominantly by the morphologic surface features of the mineral rather than by specific molecular interactions with an atomic plane of the mineral. This means that the distribution of organic molecules was controlled by surfaces like those of halite's evaporitic growth forms. The experiments with halite also demonstrated that a mineral need not continuously incorporate organic molecules during its crystallization to preserve those molecules: After rejection by (non-incorporation into) the crystallizing halite, the organic nanoparticles increased in concentration in the evaporating brine. They ultimately either adsorbed in rectilinear patterns onto the hopper-enhanced surfaces and along discontinuities within the crystals, or they were encapsulated within fluid inclusions. Of additional importance in origin-of-life considerations is the fact that halite in the natural environment rapidly can change its role from that of a protective repository (in the absence of water) to that of a source of organic particles (as soon as water is present) when the mineral dissolves.

Crystallization↗
PubMed2004

Hydrocarbon decomposition in alumina membrane: an effective way to produce carbon nanotubes bundles.

Carbon nanotubes were synthesised within the pores of an alumina membrane. The membrane had 200 nm diameter pores and 60 microm thickness, and ethylene was used as carbon source. Membrane dissolution by HF results in a bundle of parallel open tubes, aligned without macroscopic defects. The external diameter of the tubes is uniform and there is no evidence of any amorphous carbon. Wall thickness control was obtained by varying the reaction time, length by the thickness of alumina membrane, and external tube diameter by the membrane pore size. Scanning (SEM) and transmission (TEM) electron microscopy, atomic force microscopy (AFM), X-ray diffraction, thermogravimetric analysis (TG) and surface area evaluation by nitrogen adsorption were used for the characterization of membrane and nanotubes.

Adsorption↗
PubMed1999

Controlled release of human immunoglobulin G. 2. Morphological characterization.

Human immunoglobulin G (IgG) serves as an important chemotherapeutic agent for a number of immunological ailments and as a carrier in the targeted delivery of other therapeutic agents. This paper deals with the characterization of IgG-dispersed monolithic matrixes of different geometries, prepared using a nonbiodegradable polymer carrier EVAc. The morphological changes associated with the matrix during drug release was studied using scanning electron microscopy, polarizing microscopy, atomic force microscopy, and X-ray photoelectron microscopy, and the results were compared. The study answered the burst effect problem significantly and illustrated the potential of these techniques in understanding the morphological structure of matrixes and mode of release kinetics.

Delayed-Action Preparations↗
PubMed2002

Lung lining liquid modifies PM(2.5) in favor of particle aggregation: a protective mechanism.

The health effects of particle inhalation including urban air pollution and tobacco smoke comprise a significant public health concern worldwide, although the mechanisms by which inhaled particles cause premature deaths remain undetermined. In this study, we assessed the physicochemical interactions of fine airborne particles (PM(2.5)) and lung lining liquid using scanning electron microscopy, atomic force microscopy, and X-ray photon spectroscopy. We provide experimental evidence to show that lung lining liquid modifies the chemistry and attractive forces at the surface of PM(2.5), which leads to enhanced particle aggregation. We propose that this is an important protective mechanism that aids particle clearance in the lung.

Air Pollution↗
PubMed2005

Fabrication and characterization of stable ultrathin film micropatterns containing DNA and photosensitive polymer diazoresin.

Stable, ultrathin DNA micropatterns were fabricated from photosensitive polymer diazoresin (DR) through a self-assembly technique. The micropatterns were achieved on LBL ultrathin film after UV exposure through a photomask. The patterns were characterized systematically with scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy and fluorescence microscopy. All of the results indicate that the combined LBL self-assembly and photolithography technique is a promising method for constructing stable, well-defined micropatterns with a nanoscale thickness.

Animals↗
PubMed2006

Methods for assessment of dental erosion.

Various assessment techniques have been applied to evaluate the loss of dental hard tissue and the surface-softened zone in enamel induced by erosive challenges. In this chapter, the most frequently adopted techniques for analyzing the erosively altered dental hard tissues are reviewed, such as profilometry, microradiography, scanning electron microscopy, atom force microscopy, nano- and microhardness tests and iodide permeability test. Moreover, methods for chemical analysis of minerals dissolved from dental hard tissue are discussed. It becomes evident that the complex nature of erosive mineral loss and dissolution might not be comprehended by a single technique, but needs application of different approaches for full understanding.

Fluorescence↗
PubMed2007

Self-assembled molecular magnets on patterned silicon substrates: bridging bio-molecules with nanoelectronics.

The paper reports the methods of preparing molecular magnets and patterning of the molecules on a semiconductor surface. A highly magnetically aligned metallothionein containing Mn and Cd (Mn,Cd-MT-2) is first synthesized, and the molecules are then placed into nanopores prepared on silicon (001) surfaces using electron beam lithography and reactive ion-etching techniques. We have observed the self-assemble growth of the MT molecules on the patterned Si surface such that the MT molecules have grown into rod or ring type three-dimensional nanostructures, depending on the patterned nanostructures on the surface. We also provide scanning electron microscopy, atomic force microscopy, and magnetic force microscope studies of the molecular nanostructures. This engineered molecule shows molecular magnetization and is biocompatible with conventional semiconductors. These features make Mn,Cd-MT-2 a good candidate for biological applications and sensing sources of new nanodevices. Using molecular self-assembly and topographical patterning of the semiconductor substrate, we can close the gap between bio-molecules and nanoelectronics built into the semiconductor chip.

Biocompatible Materials↗
PubMed2005

Optimization of gold nanoparticle-based DNA detection for microarrays.

DNA microarrays are promising tools for fast and highly parallel DNA detection by means of fluorescence or gold nanoparticle labeling. However, substrate modification with silanes (as a prerequisite for capture DNA binding) often leads to inhomogeneous surfaces and/or nonspecific binding of the labeled DNA. We examined both different substrate cleaning and activating protocols and also different blocking strategies for optimizing the procedures, especially those for nanoparticle labeling. Contact angle measurements as well as fluorescence microscopy, atomic force microscopy (AFM), and a flatbed scanner were used to analyze the multiple-step process. Although the examined different cleaning and activating protocols resulted in considerably different contact angles, meaning different substrate wettability, silanization led to similar hydrophobic surfaces which could be revealed as smooth surfaces of about 2-4 nm roughness. The two examined silanes (3-glycidoxypropyltrimethoxysilane (GOPS) and 3-aminopropyltriethoxysilane (APTES)) differed in their DNA binding homogeneity, maximum signal intensities, and sensitivity. Nonspecific gold binding on APTES/PDC surfaces could be blocked by treatment in 3% bovine serum albumin (BSA).

Base Sequence↗
PubMed2002

Surface analysis methods for characterizing polymeric biomaterials.

Surface properties have an enormous effect on the success or failure of a biomaterial device, thus signifying the considerable importance of and the need for adequate characterization of the biomaterial surface. Microscopy techniques used in the analysis of biomaterial surfaces include scanning electron microscopy, transmission electron microscopy, atomic force microscopy, and confocal microscopy. Spectroscopic techniques include X-ray photoelectron spectroscopy, Fourier Transform infrared attenuated total reflection and secondary ion mass spectrometry. The measurement of contact angles, although one of the earlier techniques developed remains a very useful tool in the evaluation of surface hydrophobicity/hydrophilicity. This paper provides a brief, easy to understand synopsis of these and other techniques including emerging techniques, which are proving useful in the analysis of the surface properties of polymeric biomaterials. Cautionary statements have been made, numerous authors referenced and examples used to show the specific type of information that can be acquired from the different techniques used in the characterization of polymeric biomaterials surfaces.

Biocompatible Materials↗
PubMed1995

A review of the aggregation properties of a recombinant amelogenin.

The present paper reviews some recent data on the aggregation properties of a recombinant amelogenin using dynamic light scattering, transmission electron microscopy, atomic force microscopy, and size exclusion chromatography. It was found that the recombinant amelogenin M179 molecules in solution form spherical monodisperse aggregates (15-20 nm) which are predominantly stabilized by intermolecular hydrophobic interactions while their surfaces are charged, carrying the hydrophilic carboxy-terminal sequence. We concluded that the spherical aggregates represent the "stippled materials" secreted by the ameloblasts at the mineralization front. We further speculate that the self-assembly process in the formation of amelogenin aggregates may play a primary role in the structural organization of mineralizing enamel.

Agglutination↗
PubMedDate not supplied

Surface characterization of titanium-based implant materials.

This study examined the effects of different treatments (polished, electropolished, and grit-blasted) on the surface morphology and chemistry of commercially pure titanium and titanium-6% aluminum-4% vanadium. The structure and composition of the surfaces were evaluated using scanning electron microscopy, atomic force microscopy, energy dispersive spectroscopy, Auger microprobe analysis, and x-ray photoelectron spectroscopy. Surface roughness values at large scales were nearly identical for grit-blasted and electropolished samples, while at smaller scales, electropolished and polished samples had nearly identical quantitative roughness values. The surface oxide compositions were found to be primarily titanium dioxide on both materials for all surface treatments. No vanadium was seen with either x-ray photoelectron spectroscopy or Auger microprobe analysis for the alloy, indicating a possible surface depletion. Calcium was present on the grit-blasted samples, and calcium and chlorine were detected on the electropolished samples.

Alloys↗
PubMed2003

The pH dependence of HIV-1 capsid assembly and its interaction with cyclophilin A.

Immature HIV-1 virions have spherical cores which become conical due to cleavage of the capsid domain of Gag. Here, we have used an immature form of capsid and show by electron microscopy, atomic force microscopy and single angle light scattering that it aggregates to spherical cores resembling immature virions at high ionic strengths and at pH values above 6. Dynamic angle light scattering of the dissociated protein shows structural changes that promote oligomerization above pH 6. We then examined the role of the required host protein cyclophilin A on assembly. Cyclophilin A is incorporated into virions at a 1:10 cyclophilin A/capsid ratio. We find that although cyclophilin A does not affect the oligomerization rate or stability of immature capsid cores, it does bind strongly to immature capsid at physiological stoichiometry above pH 6. This association serves as an entry route of cyclophilin A into HIV-1 virions.

Capsid↗
PubMed2003

Development and characterization of novel empty adenovirus capsids and their impact on cellular gene expression.

Adenovirus (Ad) has been extensively studied as a eukaryotic viral vector. As these vectors have evolved from first-generation vectors to vectors that contain either very few or no viral genes ("gutless" Ad), significant reductions in the host innate immune response upon infection have been observed. Regardless of these vector improvements an unknown amount of toxicity has been associated with the virion structural proteins. Here we demonstrate the ability to generate high particle numbers (10(11) to 10(12)) of Ad empty virions based on a modification of Cre/lox gutless Ad vectors. Using a battery of analyses (electron microscopy, atomic force microscopy, confocal images, and competition assays) we characterized this reagent and determined that it (i) makes intact virion particles, (ii) competes for receptor binding with wild-type Ad, and (iii) enters the cell proficiently, demonstrating an ability to carry out essential steps of viral entry. To further study the biological impact of these Ad empty virions on infected cells, we carried out DNA microarray analysis. Compared to that for recombinant Ad, the number of mRNAs modulated upon infection was significantly reduced but the expression signatures were similar. This reagent provides a valuable tool for studies of Ad in that researchers can examine the effect of infection in the presence of the virion capsid alone.

Adenoviridae↗
PubMedDate not supplied

Guanidine hydrochloride can induce amyloid fibril formation from hen egg-white lysozyme.

The formation of amyloid fibrils is an intractable problem in which normally soluble protein polymerizes and forms insoluble ordered aggregates. Such aggregates can range from being a nuisance in vitro to being toxic in vivo. The latter is true for lysozyme, which has been shown to form toxic deposits in humans. In the present study, the effects of partial denaturation of hen egg-white lysozyme via incubation in a concentrated solution of the denaturant guanidine hydrochloride are investigated. Results show that when lysozyme is incubated under moderate guanidine hydrochloride concentrations (i.e., 2-5 M), where lysozyme is partially unfolded, fibrils form rapidly. Thioflavin T, Congo red, X-ray diffraction, transmission electron microscopy, atomic force microscopy, and circular dichroism spectroscopy are all used to verify the production of fibrils under these conditions. Incubation at very low or very high guanidine hydrochloride concentrations fails to produce fibrils. At very low denaturant concentrations, the structure of lysozyme is fully native and very stable. On the other hand, at very high denaturant concentrations, guanidine hydrochloride is capable of dissolving and dis-aggregating fibrils that are formed. Raising the temperature and/or concentration of lysozyme accelerates fibril formation by further adding to the concentration of partially unfolded species. The addition of preformed fibrils also accelerates fibril formation but only under partially unfolding conditions. The results presented here provide further evidence that partial unfolding is a prerequisite to fibril formation. Partial denaturation can accelerate fibril formation in much the same way that mutations have been shown to accelerate fibril formation.

Amyloid↗
PubMed2006

Carbon nanotube-DNA nanoarchitectures and electronic functionality.

Biological molecules such as deoxyribonucleic acid (DNA) possess inherent recognition and self-assembly capabilities, and are attractive templates for constructing functional hierarchical material structures as building blocks for nanoelectronics. Here we report the assembly and electronic functionality of nanoarchitectures based on conjugates of single-walled carbon nanotubes (SWNTs) functionalized with carboxylic groups and single-stranded DNA (ssDNA) sequences possessing terminal amino groups on both ends, hybridized together through amide linkages by adopting a straightforward synthetic route. Morphological and chemical-functional characterization of the nanoarchitectures are investigated using scanning electron microscopy, transmission electron microscopy, atomic force microscopy, energy-dispersive X-ray spectroscopy, Raman spectroscopy, and Fourier-transform infrared spectroscopy. Electrical measurements (I-V characterization) of the nanoarchitectures demonstrate negative differential resistance in the presence of SWNT/ssDNA interfaces, which indicates a biomimetic route to fabricating resonant tunneling diodes. I-V characterization on platinum-metallized SWNT-ssDNA nanoarchitectures via salt reduction indicates modulation of their electrical properties, with effects ranging from those of a resonant tunneling diode to a resistor, depending on the amount of metallization. Electron transport through the nanoarchitectures has been analyzed by density functional theory calculations. Our studies illustrate the great promise of biomimetic assembly of functional nanosystems based on biotemplated materials and present new avenues toward exciting future opportunities in nanoelectronics and nanobiotechnology.

DNA↗
PubMed2003

The circularization of amyloid fibrils formed by apolipoprotein C-II.

Amyloid fibrils have historically been characterized by diagnostic dye-binding assays, their fibrillar morphology, and a "cross-beta" x-ray diffraction pattern. Whereas the latter demonstrates that amyloid fibrils have a common beta-sheet core structure, they display a substantial degree of morphological variation. One striking example is the remarkable ability of human apolipoprotein C-II amyloid fibrils to circularize and form closed rings. Here we explore in detail the structure of apoC-II amyloid fibrils using electron microscopy, atomic force microscopy, and x-ray diffraction studies. Our results suggest a model for apoC-II fibrils as ribbons approximately 2.1-nm thick and 13-nm wide with a helical repeat distance of 53 nm +/- 12 nm. We propose that the ribbons are highly flexible with a persistence length of 36 nm. We use these observed biophysical properties to model the apoC-II amyloid fibrils either as wormlike chains or using a random-walk approach, and confirm that the probability of ring formation is critically dependent on the fibril flexibility. More generally, the ability of apoC-II fibrils to form rings also highlights the degree to which the common cross-beta superstructure can, as a function of the protein constituent, give rise to great variation in the physical properties of amyloid fibrils.

Amyloid↗
PubMed2003

Gas plasma etching of PEO/PBT segmented block copolymer films.

A series of poly(ethylene oxide)/poly(butylene terephthalate) (PEO/PBT) segmented block copolymer films was treated with a radio-frequency carbon dioxide (CO(2)) or with argon (Ar) plasma. The effects of (preferential) etching on surface structure, topography, chemistry, and wettability were studied by scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, and contact angle measurements. In all cases, a granular-type nanostructure was formed after prolonged CO(2) plasma etching. Ar plasma etching generally did not lead to significant changes in surface structure. Regarding surface chemistry, CO(2) plasma treatment caused surface oxidation and oxidative degradation of the films while Ar plasma etching resulted mainly in the preferential removal of PEO blocks. The wettability of all films significantly increased after plasma treatment because of the creation of polar functional groups at the surface. Preliminary goat bone-marrow cell compatibility experiments have shown that all plasma-treated PEO/PBT films induced a greatly enhanced cell adhesion and/or growth compared to untreated biomaterials. This improvement was attributed to changes in surface chemistry during plasma etching rather than to changes in surface structure. These results show that plasma-treated PEO/PBT copolymers have a high potential as scaffolds for bone tissue regeneration.

Animals↗
PubMed1999

Microstructures of an amelogenin gel matrix.

The thermo-reversible transition (clear <--> opaque) of the amelogenin gel matrix, which has been known for some three decades, has now been clarified by microstructural investigations. A mixed amelogenin preparation extracted from porcine developing enamel matrix (containing "25K," 7.4%; "23K," 10.7%; "20K," 49.5%; and smaller peptides, 32.4%) was dissolved in dilute formic acid and reprecipitated by adjusting the pH to 6.8 with NaOH solution. Amelogenin gels were formed in vitro by sedimenting the precipitate in microcentrifuge tubes. The gels were fixed with Karnovsky fixative at 4 and 24 degrees C, which was found to preserve their corresponding clear (4 degrees C) and opaque (24 degrees C) states. Scanning electron microscopy, atomic force microscopy, and transmission electron microscopy were employed for the microstructural characterization of the fixed clear and opaque gels. The amelogenin gel matrix was observed to possess a hierarchical structure of quasi-spherical amelogenin nanospheres and their assemblies. The nanospheres of diameters 8-20 nm assemble to form small spherical assemblies of diameters 40-70 nm that further aggregated to form large spherical assemblies of 70-300 nm in diameter. In the clear gel, most of the large assemblies are smaller than 150 nm, and the nanospheres and assemblies are uniformly dispersed, allowing an even fluid distribution among them. In the opaque gel, however, numerous spherical fluid-filled spaces ranging from 0.3 to 7 microm in diameter were observed with the majority of the large assemblies sized 150-200 nm in diameter. These spaces presumably result from enhanced hydrophobic interactions among nanospheres and/or assemblies as the temperature increased. The high opacity of the opaque (24 degrees C) gel apparently arises from the presence of the numerous fluid-filled spaces observed compared to the low-temperature (4 degrees C) preparation. These observations suggest that the hydrophobic interactions among nanospheres and different orders of amelogenin assemblies are important in determining the structural integrity of the dental enamel matrix.

Amelogenin↗