Biomedical subjects

C M Lieber

Publications and source records attributed to C M Lieber.

26 records · Page 2Linked to original sources

PubMed1999

Assembly of A beta amyloid protofibrils: an in vitro model for a possible early event in Alzheimer's disease.

Amyloid fibrils comprising primarily the peptides A beta 40 and A beta 42 are a defining feature of the Alzheimer's disease (AD) brain, and convergent evidence suggests that the process of their formation plays a central role in the AD pathogenic pathway. Elucidation of fibril assembly is critical for the discovery of potential AD diagnostics and therapeutics, since the pathogenic entity is not necessarily the product fibril, but could be a precursor species whose formation is linked to fibrillogenesis in vivo. Atomic force microscopy allowed the identification of an unanticipated intermediate in in vitro fibril formation, the A beta amyloid protofibril. This manuscript describes studies of the structure of the A beta 40 protofibril and its in vitro assembly and disassembly using atomic force microscopy (AFM). The A beta 40 protofibril has a height of ca. 4.3 +/- 0.5 nm and a periodicity of ca. 20 +/- 4.7 nm. The rate of its elongation depends on the total concentration of A beta 40, the temperature, and ionic strength of the medium. A beta 42 and A beta 40 protofibrils elongate at a comparable rate. Statistical analysis of AFM data reveals a decrease in the number of protofibrils with time, indicating that coalescence of smaller protofibrils contributes to protofibril elongation. Similar analysis reveals that protofibrils shorten while the number of protofibrils also decrease following dilution, indicating that protofibril disassembly does not proceed by a reverse of the assembly process. These investigations provide systematic data defining factors affecting A beta fibrillization and, thus, should be valuable in the design of high-throughput assays to identify agents which alter A beta protofibril assembly.

Adsorption↗

PubMed1998

Covalently functionalized nanotubes as nanometre-sized probes in chemistry and biology.

Carbon nanotubes combine a range of properties that make them well suited for use as probe tips in applications such as atomic force microscopy (AFM). Their high aspect ratio, for example, opens up the possibility of probing the deep crevices that occur in microelectronic circuits, and the small effective radius of nanotube tips significantly improves the lateral resolution beyond what can be achieved using commercial silicon tips. Another characteristic feature of nanotubes is their ability to buckle elastically, which makes them very robust while limiting the maximum force that is applied to delicate organic and biological samples. Earlier investigations into the performance of nanotubes as scanning probe microscopy tips have focused on topographical imaging, but a potentially more significant issue is the question of whether nanotubes can be modified to create probes that can sense and manipulate matter at the molecular level. Here we demonstrate that nanotube tips with the capability of chemical and biological discrimination can be created with acidic functionality and by coupling basic or hydrophobic functionalities or biomolecular probes to the carboxyl groups that are present at the open tip ends. We have used these modified nanotubes as AFM tips to titrate the acid and base groups, to image patterned samples based on molecular interactions, and to measure the binding force between single protein-ligand pairs. As carboxyl groups are readily derivatized by a variety of reactions, the preparation of a wide range of functionalized nanotube tips should be possible, thus creating molecular probes with potential applications in many areas of chemistry and biology.

Biology↗

PubMed1997

Observation of metastable Abeta amyloid protofibrils by atomic force microscopy.

BACKGROUND: Brain amyloid plaque, a diagnostic feature of Alzheimer's disease (AD), contains an insoluble fibrillar core that is composed primarily of variants of the beta-amyloid protein (Abeta). As Abeta amyloid fibrils may initiate neurodegeneration, the inhibition of fibril formation is a possible therapeutic strategy. Very little is known about the early steps of the process, however. RESULTS: Atomic force microscopy was used to follow amyloid fibril formation in vitro by the Abeta variants Abeta1-40 and Abeta1-42. Both variants first form small ordered aggregates that grow slowly and then rapidly disappear, while prototypical amyloid fibrils of two discrete morphologies appear. Abeta1-42 aggregates much more rapidly than Abeta1-40, which is consistent with its connection to early-onset AD. We propose that the metastable intermediate species be called Abeta amyloid protofibrils. CONCLUSIONS: Abeta protofibrils are likely to be intermediates in the in vitro assembly of Abeta amyloid fibrils, but their in vivo role has yet to be determined. Numerous reports of a nonfibrillar form of Abeta aggregate in the brains of individuals who are predisposed to AD suggest the existence of a precursor form, possibly the protofibril. Thus, stabilization of Abeta protofibrils may be a useful therapeutic strategy.

Alzheimer Disease↗

PubMed1997

Atomic force microscopic imaging of seeded fibril formation and fibril branching by the Alzheimer's disease amyloid-beta protein.

BACKGROUND: Amyloid plaques composed of the fibrillar form of the amyloid-beta protein (Abeta) are the defining neuropathological feature of Alzheimer's disease (AD). A detailed understanding of the time course of amyloid formation could define steps in disease progression and provide targets for therapeutic intervention. Amyloid fibrils, indistinguishable from those derived from an AD brain, can be produced in vitro using a seeded polymerization mechanism. In its simplest form, this mechanism involves a cooperative transition from monomeric Abeta to the amyloid fibril without the buildup of intermediates. Recently, however, a transient species, the Abeta amyloid protofibril, has been identified. Here, we report studies of Abeta amyloid protofibril and its seeded transition into amyloid fibrils using atomic force microscopy. RESULTS: Seeding of the protofibril-to-fibril transition was observed. Preformed fibrils, but not protofibrils, effectively seeded this transition. The assembly state of Abeta influenced the rate of seeded growth, indicating that protofibrils are fibril assembly precursors. The handedness of the helical surface morphology of fibrils depended on the chirality of Abeta. Finally, branched and partially wound fibrils were observed. CONCLUSIONS: The temporal evolution of morphologies suggests that the protofibril-to-fibril transition is nucleation-dependent and that protofibril winding is involved in that transition. Fibril unwinding and branching may be essential for the post-nucleation growth process. The protofibrillar assembly intermediate is a potential target for AD therapeutics aimed at inhibiting amyloid formation and AD diagnostics aimed at detecting presymptomatic disease.

Alzheimer Disease↗

PubMed1997

Stretching and breaking duplex DNA by chemical force microscopy.

BACKGROUND: Specific interactions between complementary strands of DNA and other molecules are central to the storage, retrieval and modification of information in biological systems. Although in many cases the basic structures of duplex DNA and the binding energetics have been well characterized, little information is available about the forces in these systems. These forces are of critical importance because they must be overcome, for example, by protein machines during transcription and repair. Recent developments in atomic force microscopy make possible direct measurements of such forces between the individual oligonucleotide strands that form DNA duplexes. RESULTS: We used the chemical force microscopy technique, in which oligonucleotides are covalently linked to the force microscope probe tip and the sample surface, to measure the elongation and binding forces of individual DNA duplexes. The separation forces between complementary oligonucleotide strands were found to be significantly larger than the forces measured between noncomplementary strands, and to be consistent with the unbinding of a single DNA duplex. With increasing applied force, the separation of complementary strands proceeded in a stepwise manner: B-form DNA was stretched, then structurally transformed to a stable form of DNA approximately twice the length of the B form, and finally separated into single-stranded oligonucleotides. These data provide a direct measurement of the forces required to elastically deform and separate double-stranded DNA into single strands. CONCLUSIONS: Force microscopy provides a direct and quantitative measurement of the forces and energetics required to stretch and unbind DNA duplexes. Because the measurements can be carried out readily on synthetic oligonucleotides and in the presence of exogenous molecules, this method affords an opportunity for directly assessing the energetics of distorting and unbinding specific DNA sequences and DNA complexes. Such data could provide unique insights into the mechanistic steps following sequence-specific recognition by, for example, DNA repair and transcription factors.

DNA↗

PubMed1995

Time required to remove totally bonded tooth-colored posterior restorations and related tooth substance loss.

OBJECTIVES: The study was conducted to measure the time required to remove large totally bonded tooth-colored posterior restorations and related tooth substance loss. This information was collected to determine if there were differences between bonded restorations and conventional restorations. METHODS: Molars were restored with the following materials: amalgam, composite, glass ionomer cement or glass ceramic cusp coverages. After submitting them to an in vitro aging process, they were attached to a lower jaw model in a phantom head. Six dentists removed the test restorations under standardized, quasi-clinical conditions. RESULTS: While the amalgam was completely removed, some glass ionomer cement vestiges were found. In the glass-ceramic group, the margins were covered with remnants of the composite luting agent in several places. The most restorative material was left in the composite group. The loss of tooth structure after removal of amalgam, glass ionomer cement, composite and glass ceramic was 17.6 mm3, 19.6 mm3, 39.9 mm3 and 41.8 mm3, respectively. When comparing the removal time, the glass ionomer group scored best with an average of 11.9 min followed by the amalgam group with an average of 15.2 min, followed by 24.9 min for the composite group and 30.4 min for the glass ceramic group. SIGNIFICANCE: The removal of totally bonded posterior restorations made of composite and ceramic is more technically demanding and more time-consuming than the removal of glass ionomer and amalgam restorations.

Acrylic Resins↗

PubMed1991

Scanning tunneling microscopy imaging of synthetic oligonucleotides and oligonucleotide-metal complexes.

Scanning tunneling microscopy (STM) has been used to image 12- and 24-base pair (bp) synthetic oligonucleotide duplexes alone or with intercalatively-bound metal complexes with submolecular resolution. The sizes of the 12- and 24-bp oligonucleotides determined from STM images are close to expected values, and images of isolated duplexes resolve the two nucleotide strands of these molecules. The variation of duplex size in our images demonstrates that these features are not due to surface artifacts. In addition, images of the 12-bp duplex in the presence of bis(9,10-phenanthrenequinone diimine)(2,2'-bipyridyl)rhodium(III) exhibit a new structural feature at 14 +/- 2 A from the duplex ends. This new feature corresponds well to the metal binding site determined from DNA cleavage and molecular modeling studies. These results indicate that STM can be used to image directly transition metal complexes bound to DNA, and thus suggest that metal complexes bound specifically to biological molecules could serve as labels in STM structural studies.

2,2'-Dipyridyl↗

PubMed1990

Enhancement of the critical current density in single-crystal Bi2Sr2CaCu2O8 superconductors by chemically induced disorder.

The effect of metal substitution on the critical current densities of single-crystal PbxBi2-xSr2CaCu2O8 (x = 0 or x = 0.7) superconductors has been investigated. Substitution of lead was found to increase the average critical current density from 1 x 10(5) A/cm2 to 2 x 10(6) A/cm2 at 5 K in an applied magnetic field of 10 kilooersteds (1 oersted = 80 A/m). The order of magnitude increase in the critical current density was observed for temperatures up to the flux vortex lattice melting point; the flux lattice melting point was also found to increase to 30 K (from 22 K) in the lead-substituted materials. Diffraction and microscopy investigations of the structural parameters indicate that the fundamental atomic lattices are virtually the same for both materials. Scanning tunneling microscopy images demonstrate, however, that lead substitution causes significant disorder (or defects) in the one-dimensional superstructure found in Bi2Sr2CaCu2O8. Since crystal defects can increase the critical current density by pinning the motion of flux vortices, it is likely that this lead-induced disorder enhances vortex pinning. The lead-induced disorder is specific to the nonsuperconducting Bi-O layers, and thus our results suggest that chemical substitutions may be utilized to control selectively flux pinning and the critical current density in these materials.

Journal Article↗