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Biomedical subjects

James W Brown

Publications and source records attributed to James W Brown.

13 recordsLinked to original sources

The RNA Ontology Consortium: an open invitation to the RNA community.

The aim of the RNA Ontology Consortium (ROC) is to create an integrated conceptual framework-an RNA Ontology (RO)-with a common, dynamic, controlled, and structured vocabulary to describe and characterize RNA sequences, secondary structures, three-dimensional structures, and dynamics pertaining to RNA function. The RO should produce tools for clear communication about RNA structure and function for multiple uses, including the integration of RNA electronic resources into the Semantic Web. These tools should allow the accurate description in computer-interpretable form of the coupling between RNA architecture, function, and evolution. The purposes for creating the RO are, therefore, (1) to integrate sequence and structural databases; (2) to allow different computational tools to interoperate; (3) to create powerful software tools that bring advanced computational methods to the bench scientist; and (4) to facilitate precise searches for all relevant information pertaining to RNA. For example, one initial objective of the ROC is to define, identify, and classify RNA structural motifs described in the literature or appearing in databases and to agree on a computer-interpretable definition for each of these motifs. To achieve these aims, the ROC will foster communication and promote collaboration among RNA scientists by coordinating frequent face-to-face workshops to discuss, debate, and resolve difficult conceptual issues. These meeting opportunities will create new directions at various levels of RNA research. The ROC will work closely with the PDB/NDB structural databases and the Gene, Sequence, and Open Biomedical Ontology Consortia to integrate the RO with existing biological ontologies to extend existing content while maintaining interoperability.

Databases, Genetic↗

Structural implications of novel diversity in eucaryal RNase P RNA.

Previous eucaryotic RNase P RNA secondary structural models have been based on limited diversity, representing only two of the approximately 30 phylogenetic kingdoms of the domain Eucarya. To elucidate a more generally applicable structure, we used biochemical, bioinformatic, and molecular approaches to obtain RNase P RNA sequences from diverse organisms including representatives of six additional kingdoms of eucaryotes. Novel sequences were from acanthamoeba (Acathamoeba castellanii, Balamuthia mandrillaris, Filamoeba nolandi), animals (Caenorhabditis elegans, Drosophila melanogaster), alveolates (Theileria annulata, Babesia bovis), conosids (Dictyostelium discoideum, Physarum polycephalum), trichomonads (Trichomonas vaginalis), microsporidia (Encephalitozoon cuniculi), and diplomonads (Giardia intestinalis). An improved alignment of eucaryal RNase P RNA sequences was assembled and used for statistical and comparative structural analysis. The analysis identifies a conserved core structure of eucaryal RNase P RNA that has been maintained throughout evolution and indicates that covariation in size occurs between some structural elements of the RNA. Eucaryal RNase P RNA contains regions of highly variable length and structure reminiscent of expansion segments found in rRNA. The eucaryal RNA has been remodeled through evolution as a simplified version of the structure found in bacterial and archaeal RNase P RNAs.

Acanthamoeba castellanii↗

Bartonella quintana in cynomolgus monkey (Macaca fascicularis).

We identified a Bartonella quintana strain by polymerase chain reaction amplification, cloning, and sequencing of DNA extracted from lysed erythrocytes and cultured colonies grown from peripheral blood collected from a captive-bred cynomolgus monkey (Macaca fascicularis). This report describes naturally acquired B. quintana infection in a nonhuman primate.

Animals↗

Above-water radiometry in shallow coastal waters.

Above- and in-water radiometric data were collected from two coastal platforms: a small boat and an oceanographic tower. The above-water data were processed with and without a correction for bidirectional effects (Q02 and S95, respectively). An intercomparison of water-leaving radiances over a wide range of environmental conditions showed (a) total uncertainties across the blue-green domain were to within 4%, (b) a convergence of the Q02 method with the in-water method (average Q02 intercomparisons were to within 4%), and (c) chlorophyll a concentrations derived from Q02 reflectances and the OC4V4 (Ocean Color 4 Version 4) algorithm agreed with independent high-performance liquid-chromatography determinations to within approximately 32%.

Journal Article↗

Interactions between RNase P protein subunits in archaea.

A yeast two-hybrid system was used to identify protein-protein interactions between the ribonuclease P (RNase P) protein subunits Mth11p, Mth687p, Mth688p and Mth1618p from the archaeon Methanothermobacter thermoautotrophicus. Clear interactions between Mth688p and Mth687p, and between Mth1618p and Mth11p, were confirmed by HIS3 and LacZ reporter expression. Weaker interactions of Mth687p and Mth688p with Mth 11p, and Mth11p with itself, are also suggested. These interactions resemble, and confirm, those previously seen among the homologs of these proteins in the more complex yeast RNase P holoenzyme.

Methanobacteriaceae↗

In vitro selection of an archaeal RNase P RNA mimics natural variation.

Archaeal and bacterial RNase P RNAs are similar in sequence and secondary structure, but in the absence of protein, the archaeal RNAs are much less active and require extreme ionic conditions for activity. To assess how readily the activity of the archaeal RNA alone could be improved by small changes in sequence, in vitro selection was used to generate variants of a Methanobacterium formicicum RNase P RNA: Bacillus subtilus pre-tRNA(Asp) self-cleaving conjugate RNA. Functional variants were generated with a spectrum of mutations that were predominately consistent with natural variation in this RNA. Variants generated from the selection had cleavage rates comparable to that of wild type; variants with improved cleavage rates or lower ionic requirements were not obtained. This suggests that the RNase P RNAs of Bacteria and Archaea are globally optimized and the basis for the large biochemical differences between these two types of RNase P RNA is distributed in the molecule.

Archaea↗

Genes within genes within bacteria.

Recently, an unusual gene structure has been described in species of the genus Thermus, in which the rpmH (ribosomal protein L34) coding sequence was found to be entirely overlapped by the unusually large rnpA (RNase P protein subunit) sequence. Gene overlap is common in viruses, but has not been seen to this extent in any bacterium.

Bacteria↗

RNAML: a standard syntax for exchanging RNA information.

Analyzing a single data set using multiple RNA informatics programs often requires a file format conversion between each pair of programs, significantly hampering productivity. To facilitate the interoperation of these programs, we propose a syntax to exchange basic RNA molecular information. This RNAML syntax allows for the storage and the exchange of information about RNA sequence and secondary and tertiary structures. The syntax permits the description of higher level information about the data including, but not restricted to, base pairs, base triples, and pseudoknots. A class-oriented approach allows us to represent data common to a given set of RNA molecules, such as a sequence alignment and a consensus secondary structure. Documentation about experiments and computations, as well as references to journals and external databases, are included in the syntax. The chief challenge in creating such a syntax was to determine the appropriate scope of usage and to ensure extensibility as new needs will arise. The syntax complies with the eXtensible Markup Language (XML) recommendations, a widely accepted standard for syntax specifications. In addition to the various generic packages that exist to read and interpret XML formats, an XML processor was developed and put in the open-source MC-Core library for nucleic acid and protein structure computer manipulation.

Databases, Nucleic Acid↗

Archaeal RNase P has multiple protein subunits homologous to eukaryotic nuclear RNase P proteins.

Although archaeal RNase P RNAs are similar in both sequence and structure to those of Bacteria rather than eukaryotes, and heterologous reconstitution between the Bacillus subtilis RNase P protein and some archaeal RNase P RNAs has been demonstrated, no archaeal protein sequences with similarity to any known bacterial RNase P protein subunit have been identified, and the density of Methanothermobacter thermoautotrophicus RNase P in Cs2SO4 (1.42 g/mL) is inconsistent with a single small bacterial-like protein subunit. Four hypothetical open reading frames (MTH11, MTH687, MTH688, and MTH1618) were identified in the genome of M. thermoautotrophicus that have sequence similarity to four of the nine Saccharomyces cerevisiae RNase P protein subunits: Pop4p, Pop5p, Rpp1p, and Rpr2p, respectively. Polyclonal antisera generated to recombinant Mth11p, Mth687p, Mth688p, and Mth1618p each recognized a protein of the predicted molecular weight in western blots of partially purified M. thermoautotrophicus RNase P, and immunoprecipitated RNase P activity from the same partially purified preparation. RNase P in Archaea is therefore composed of an RNA subunit similar to bacterial RNase P RNA and multiple protein subunits similar to those in the eukaryotic nucleus.

Amino Acid Sequence↗

Investigation of the phylogenetic relationships within the genus Bartonella based on comparative sequence analysis of the rnpB gene, 16S rDNA and 23S rDNA.

A variety of genes and analytical methods have been applied to the study of phylogenetic relationships within the genus Bartonella, but so far the results have been inconsistent. While previous studies analysed single protein-encoding genes, we have analysed an alignment containing the sequences of three important phylogenetic markers, RNase P RNA, 16S rRNA and 23S rRNA, merged by catenation, to determine the phylogenetic relationships within the genus Bartonella. The dataset described here comprises 13 different Bartonella strains, including the seven strains that are known to be human pathogens. A variety of algorithms have been used to construct phylogenetic trees based on the combined alignment and, for comparison purposes, each individual gene. Trees generated from the catenated alignment are more consistent (independent of algorithm) and robust (higher bootstrap support). It is suggested that a phylogenetic analysis incorporating the RNase P RNA, 16S rRNA and 23S rRNA be used to study the phylogenetic relationships within the genus Bartonella.

Algorithms↗

Radiopacity of nonmetallic root canal posts.

PURPOSE: This study determined the radiopacity of a group of nonmetallic posts. MATERIALS AND METHODS: Four specimens were cut from 7 posts, and tooth sections were cut from extracted teeth. Radiographic images of all specimens along with an aluminum step wedge were obtained on occlusal films. Optical density readings for each specimen image were determined with a transmission densitometer. Radiopacity values were subsequently calculated as equivalents of aluminum thickness. RESULTS: Analysis of variance revealed significant differences in radiopacity values among the posts (P < .001). One nonmetallic post that was made of zirconium had a radiopacity value significantly greater than that of enamel. Another, made of glass fibers, acrylic resin, and fillers, had a radiopacity that was greater than that of dentin but smaller than that of enamel. The remaining 3 nonmetallic posts had radiopacity values lower than dentin. CONCLUSION: The 3 posts with radiopacity that was lower than dentin cannot be considered sufficiently radiopaque.

Absorptiometry, Photon↗