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At least 433 records · Page 24Linked to original sources

PubMed1998

A genetic algorithm for designing gene family-specific oligonucleotide sets used for hybridization: the G protein-coupled receptor protein superfamily.

MOTIVATION: Massive oligonucleotide hybridization is one of the most promising technologies of functional genome analysis. The critical point is to design appropriate sets of oligonucleotides that can be used effectively in identification by hybridization. RESULTS: Using a genetic algorithm approach, we have attempted to design sets of oligo probes capable of identifying new genes belonging to a defined gene family within a cDNA or genomic library. It is not limited by oligonucleotide length and admits the letter 'N' in the structure of the oligonucleotides selected. One of the major advantages of this approach is the low homology required to identify functional families of sequences with little homology. We have designed the oligonucleotide sets that are most selective for the cDNA clones of transmembrane G protein-coupled receptors (GPCRs), a large family of proteins that form part of a modular system of extracellular signal transduction to the intracellular second messenger pathways. The accuracy of identification has been checked on the EST library containing 713 870 cDNA sequences. A set of 15 oligos between 7 and 14 bases in length has correctly identified 70% of the GPCR cDNA collection sequences with 0.02% false positives. AVAILABILITY: The developed software is available by ftp://ftp.bionet.nsc. ru/pub/biology/ and on the Web page http://www.bionet.nsc. ru/SRCG/Oligoselector/. CONTACT: kel@.bionet.nsc.ru; sebastian. meier-ewert@gpc-ag.com

Algorithms↗

PubMed2003

An ordered collection of expressed sequences from Cryphonectria parasitica and evidence of genomic microsynteny with Neurospora crassa and Magnaporthe grisea.

Cryphonectria parasitica, the causative agent of chestnut blight, has proven to be a tractable experimental system for studying fungal pathogenesis. Moreover, the development of infectious cDNA clones of C. parasitica hypoviruses, capable of attenuating fungal virulence, has provided the opportunity to examine molecular aspects of fungal plant pathogenesis in the context of biological control. In order to establish a genomic base for future studies of C. parasitica, the authors have analysed a collection of expressed sequences. A mixed cDNA library was prepared from RNA isolated from wild-type (virus-free) and hypovirus-infected C. parasitica strains. Plasmid DNA was recovered from individual transformants and sequenced from the 5' end of the insert. Contig analysis of the collected sequences revealed that they represented approximately 2200 individual ORFs. An assessment of functional diversity present in this collection was achieved by using the BLAST software utilities and the NCBI protein database. Candidate genes were identified with significant potential relevance to C. parasitica growth, development, pathogenesis and vegetative incompatibility. Additional investigations of a 12.9 kbp genomic region revealed microsynteny between C. parasitica and both Neurospora crassa and Magnaporthe grisea, two closely related fungi. These data represent the largest collection of sequence information currently available for C. parasitica and are now forming the basis of further studies using microarray analyses to determine global changes in transcription that occur in response to hypovirus infection.

Amino Acid Sequence↗

PubMed2004

Preparation of single rice chromosome for construction of a DNA library using a laser microbeam trap.

We report the development of a laser micromanipulation system and its application in the isolation of individual rice chromosomes directly from a metaphase cell. Microdissection and flow sorting are two major methods for the isolation of single chromosome. These methods are dependent on the techniques of chromosome spread and chromosome suspension, respectively. In the development of this system, we avoided using chromosome spread and cell suspension was used instead. The cell wall of metaphase rice cell was cut by optical scissors. The released single chromosome was captured by an optical trap and transported to an area without cell debris. The isolated single chromosome was then collected and specific library was constructed by linker adaptor PCR. The average insert size of the library was about 300 bp. Two hundred inserts of chromosome 4 library were sequenced, and 96.5% were aligned to the corresponding sequences of rice chromosome 4. These results suggest the possible application of this method for the preparation of other subcellular structures and for the cloning of single macromolecule through a laser microbeam trap.

Chromosomes, Plant↗

PubMed2003

The Association of Academic Health Sciences Libraries Annual Statistics: an exploratory twenty-five-year trend analysis.

This paper presents an exploratory trend analysis of the statistics published over the past twenty-four editions of the Annual Statistics of Medical School Libraries in the United States and Canada. The analysis focuses on the small subset of nineteen consistently collected data variables (out of 656 variables collected during the history of the survey) to provide a general picture of the growth and changing dimensions of services and resources provided by academic health sciences libraries over those two and one-half decades. The paper also analyzes survey response patterns for U.S. and Canadian medical school libraries, as well as osteopathic medical school libraries surveyed since 1987. The trends show steady, but not dramatic, increases in annual means for total volumes collected, expenditures for staff, collections and other operating costs, personnel numbers and salaries, interlibrary lending and borrowing, reference questions, and service hours. However, when controlled for inflation, most categories of expenditure have just managed to stay level. The exceptions have been expenditures for staff development and travel and for collections, which have both outpaced inflation. The fill rate for interlibrary lending requests has remained steady at about 75%, but the mean ratio of items lent to items borrowed has decreased by nearly 50%.

Annual Reports as Topic↗

PubMed2006

Scopus database: a review.

The Scopus database provides access to STM journal articles and the references included in those articles, allowing the searcher to search both forward and backward in time. The database can be used for collection development as well as for research. This review provides information on the key points of the database and compares it to Web of Science. Neither database is inclusive, but complements each other. If a library can only afford one, choice must be based in institutional needs.

Journal Article↗

PubMed2006

Phenotypic activation to discover biological pathways and kinase substrates.

Observing the effects of gene perturbation on cells or organisms has long been a standard strategy in biological research. We developed a genome-wide gene overexpression library as a new tool for large-scale functional analysis in budding yeast. Previous large-scale genetic studies have focused on applications of the deletion mutant collection, which has arguably revolutionized the functional characterization of yeast genes. While extremely powerful, deletion mutant experiments are generally limited to the characterization of loss-of-function phenotypes. We have explored the potential for using the Synthetic Genetic Array (SGA) method, a platform for high-throughput genetic analysis, with a genome-wide "overexpression array", in which each strain on the array overexpresses a unique yeast gene. The overexpression array enables gain-of-function phenotypes to be examined on a large scale, providing a unique insight into gene function and a novel source of reagents for the global mapping of genetic networks and functional relationships amongst genes and pathways. Understanding the molecular bases of overexpression phenotypes should also shed new light on the nature of genetic dominance.

Animals↗

PubMed1994

A unique approach to multi-state networking: BHSL (Basic Health Sciences Network).

Development of a reciprocal multi-state shared resources network is described. The Basic Health Sciences Library Network (BHSL) is one the largest interlibrary loan networks free of direct charges to participants and any direct federal or state funding. Established in June 1986, BHSL started with 132 member libraries from three northeastern states. Current membership is 460 libraries in 10 states. Interlibrary loan activity for 1992 resulted in a collective cost savings of $592,672. This model of resource sharing can be applied to any group of libraries that access a common locator tool.

Cost Savings↗

PubMed2001

REALISE-ing their potential?: Implementing local library projects to support evidence-based health care.

Librarian involvement in Evidence-based Health Care provides many opportunities at a local level. Unfortunately, the potential for innovative projects to inform future developments is generally lost by a failure to 'pass the baton'--to identify lessons learnt and transferable principles. The 'Library Support for Evidence-based Health Care' Project, funded by the NHS Executive Northern and Yorkshire, resulted in the implementation of locally responsive packages of hardware and software in six of the Region's libraries. The opportunity to evaluate the collective experience of these sites, and to synthesize principles of good practice, was provided by a separately funded post-hoc evaluation, the Research Evaluation to Audit Library and Information Support for EBHC (REALISE). This paper reports on how this evaluation was conducted, documents the strengths and weaknesses of the Project itself, and attempts to provide a checklist for use in similar projects. The paper concludes by outlining the relevance of the findings to the introduction of planned organizational approaches to quality (clinical governance) and the development of local implementation strategies across the UK, required by the NHS Information Strategy, Information for Health.

Evaluation Studies as Topic↗

PubMed2006

Professionally applied topical fluoride: evidence-based clinical recommendations.

BACKGROUND: With the dramatic increase in the amount of scientific information available about oral health, an evidence-based approach to oral health care and the practice of dentistry is necessary. There is a need to summarize, critique and disseminate scientific evidence and to translate the evidence into a practical format that is used easily by dentists The evidence-based clinical recommendations in this report were developed by an expert panel established by the American Dental Association Council on Scientific Affairs that evaluated the collective body of scientific evidence on the effectiveness of professionally applied topical fluoride for caries prevention. The recommendations are intended to assist dentists in clinical decision making. TYPES OF STUDIES REVIEWED: MEDLINE and the Cochrane Library were searched for systematic reviews and clinical studies of professionally applied topical fluoride--including gel, foam and varnish--through October 2005. RESULTS: Panelists were selected on the basis of their expertise in the relevant subject matter. The recommendations are stratified by age groups and caries risk and indicate that periodic fluoride treatments should be considered for both children and adults who are at moderate or high risk of developing caries. Included in the clinical recommendations is a summary table that can be used as a chairside resource. CLINICAL IMPLICATIONS: The dentist, knowing the patient's health history and vulnerability to oral disease, is in the best position to make treatment decisions in the interest of each patient. These clinical recommendations must be balanced with the practitioner's professional expertise and the individual patient's preferences.

Adolescent↗

PubMed2002

The Italian Twin Project: from the personal identification number to a national twin registry.

The unique opportunity given by the "fiscal code", an alphanumeric identification with demographic information on any single person residing in Italy, introduced in 1976 by the Ministry of Finance, allowed a database of all potential Italian twins to be created. This database contains up to now name, surname, date and place of birth and home address of about 1,300,000 "possible twins". Even though we estimated an excess of 40% of pseudo-twins, this still is the world's largest twin population ever collected. The database of possible twins is currently used in population-based studies on multiple sclerosis, Alzheimer's disease, celiac disease, and type 1 diabetes. A system is currently being developed for linking the database with data from mortality and cancer registries. In 2001, the Italian Government, through the Ministry of Health, financed a broad national research program on twin studies, including the establishment of a national twin registry. Among all the possible twins, a sample of 500,000 individuals are going to be contacted and we expect to enrol around 120,000 real twin pairs in a formal Twin Registry. According to available financial resources, a sub sample of the enrolled population will be asked to donate DNA. A biological bank from twins will be then implemented, guaranteeing information on future etiological questions regarding genetic and modifiable factors for physical impairment and disability, cancers, cardiovascular diseases and other age related chronic illnesses.

Aging↗

PubMed1980

MEDLINE: an introduction to on-line searching.

MEDLINE is presented as a prototype for on-line bibliographic search systems. Creation of the data base, indexing language, and file organization are reviewed. On accessing the files, search logic is illustrated with a sample MEDLINE search. NLM's development of a document delivery system to complement its bibliographic retrieval system is discussed.

Abstracting and Indexing↗

PubMed2003

Dereplication of Streptomyces soil isolates and detection of specific biosynthetic genes using an automated ribotyping instrument.

The discrimination of distinct cultures among morphologically similar Streptomyces soil isolates (dereplication) and the detection of specific biosynthetic pathways in these strains are important steps in the selection of microorganisms to include in a natural products library. We have developed methods for analysis of actinomycetes using the RiboPrinter microbial characterization system, an automated instrument that performs ribotyping on bacterial samples. To evaluate our dereplication method, 26 Streptomyces isolates, obtained from soil samples collected in Maui, Hawaii, were ribotyped and compared with each other, using the RiboPrinter. The strains were also compared by 16S rDNA sequence analysis, MIDI fatty acid analysis, and LC-MS profiling of fermentation extracts. The RiboPrinter was able to identify closely related isolates and to discriminate between morphologically similar isolates with unique genetic, fatty acid and fermentation profiles. For the detection of biosynthetic genes, a 1,006-bp probe containing a portion of an adenylation domain of a non-ribosomal peptide synthetase (NRPS) was employed. Using this alternate probe in place of the standard ribosomal probe, the RiboPrinter was able to detect NRPS genes in several strains of Streptomyces. These results demonstrate that the RiboPrinter has multiple applications in a natural products research program.

Biotechnology↗

PubMed2005

Real-time protein kinase assay.

We report a novel, real-time fluorogenic kinase assay. The peptide substrates are synthesized with a fluorescent dye and a hydrocarbon tail. The substrate self-assembles into micelles, increasing the local concentration of the dye and quenching its fluorescence. Upon phosphorylation, the fluorescence intensity increases 4-6-fold due to micelle reorganization. Both dynamic light scattering data and cryoelectron microscope images show that the size and the shape of the phosphopeptide micelles are significantly different from micelles of substrate peptide. The system provides a robust fluorescence increase in a real-time protein kinase assay. Unlike other fluorogenic systems, the fluorophore may be distant from the serine, threonine, or tyrosine that is phosphorylated. Assays for several kinases, including PKA, PKC, p38, MAPKAP K2, akt, Erk1, and src-family kinases, have been developed. IC(50) values of inhibitors for PKC betaII determined with this technology are consistent with published values. The utility of this assay to high-throughput screening was demonstrated with Sigma's LOPAC library, a collection of 640 compounds with known biological activities, and satisfactory results were obtained.

Adenosine Triphosphate↗

PubMedDate not supplied

Molecular shape diversity of combinatorial libraries: a prerequisite for broad bioactivity.

A computational method to rapidly assess and visualize the diversity in molecular shape associated with a given compound set has been developed. Normalized ratios of principal moments of inertia are plotted into two-dimensional triangular graphs and then used to compare the shape space covered by different compound sets, such as combinatorial libraries of varying size and composition. We have further developed a computational method to analyze interset similarity in terms of shape space coverage, which allows the shape redundancy between the different subsets of a given compound collection to be analyzed in a quantitative way. The shape space coverage has been found to originate mainly from the nature and the 3D-geometry (but not the size) of the central scaffold, while the number and nature of the peripheral substituents and conformational aspects were shown to be of minor importance. Substantial shape space coverage has been correlated with broad biological activity by applying the same shape analysis to collections of known bioactive compounds, such as MDDR and the GOLD-set. The aggregate of our results corroborates the intuitive notion that molecular shape is intimately linked to biological activity and that a high degree of shape (hence scaffold) diversity in screening collections will increase the odds of addressing a broad range of biological targets.

Combinatorial Chemistry Techniques↗

PubMed1997

A collection of 1814 human chromosome 7-specific STSs.

An established goal of the ongoing Human Genome Project is the development and mapping of sequence-tagged sites (STSs) every 100 kb, on average, across all human chromosomes. En route to constructing such a physical map of human chromosome 7, we have generated 1814 chromosome 7-specific STSs. The corresponding PCR assays were designed by the use of DNA sequence determined in our laboratory (79%) or generated elsewhere (21%) and were demonstrated to be suitable for screening yeast artificial chromosome (YAC) libraries. This collection provides the requisite landmarks for constructing a physical map of chromosome 7 at < 100-kb average spacing of STSs.

Base Sequence↗

PubMed2004

Binding proteins from alternative scaffolds.

The use of so-called protein scaffolds for the generation of novel binding proteins via combinatorial engineering has recently emerged as a powerful alternative to natural or recombinant antibodies. This concept requires an extraordinary stable protein architecture tolerating multiple substitutions or insertions at the primary structural level. With respect to broader applicability it should involve a type of polypeptide fold which is observed in differing natural contexts and with distinct biochemical functions, so that it is likely to be adaptable to novel molecular recognition purposes. The quickly growing number of approaches can be classified into three groups: carrier proteins for the display of single variegated loops, scaffolds providing rigid elements of secondary structure, and protein frameworks supporting a group of conformationally variable loops in a fixed spatial arrangement. Generally, such artificial receptor proteins should be based on monomeric and small polypeptides that are robust, easily engineered, and efficiently produced in inexpensive prokaryotic expression systems. Today, progress in protein library technology allows for the parallel development of immunoglobulin (Ig) as well as scaffold-based affinity reagents. Both biomolecular tools have the potential to complement each other, thus expanding the possibility to find an affinity reagent suitable for a given application. The repertoire of protein scaffolds hitherto recruited for combinatorial protein engineering purposes will probably be further expanded in the future, including both additional natural proteins and de novo designed proteins, contributing to the collection of libraries available at present. In this review both the structural features and the practical use of scaffold proteins will be discussed and exemplified.

Animals↗

PubMed2006

DNA as a dietary biomarker in Antarctic krill, Euphausia superba.

The diet of Antarctic krill (Euphausia superba) has been studied using a variety of techniques, but current methods still suffer from problems that are difficult to solve. This study examined an alternative approach utilizing DNA as a prey biomarker. Methods were developed for the preservation, extraction, and identification of prey DNA from krill collected in the field. Group-specific polymerase chain reaction (PCR) was used to amplify diatom prey (Phylum: Bacillariophyta) and the results from DNA clone libraries were compared with microscopic diet analysis. DNA analysis was superior to microscopy for prey detection. However, differences in prey relative abundance estimates between the two techniques suggested some bias in the DNA-based estimates. Quantification showed that large amounts of prey DNA had been successfully preserved and extracted. Overall the results suggest that the application of DNA-based diet analysis to krill warrants further investigation, particularly for prey that are difficult to study using other methods.

Animals↗

PubMed2006

HepSEQ: International Public Health Repository for Hepatitis B.

HepSEQ is a repository for an extensive library of public health and molecular data relating to hepatitis B virus (HBV) infection collected from international sources. It is hosted by the Centre for Infections, Health Protection Agency (HPA), England, United Kingdom. This repository has been developed as a web-enabled, quality-controlled database to act as a tool for surveillance, HBV case management and for research. The web front-end for the database system can be accessed from http://www.hpa-bioinfodatabases.org.uk/hepatitis_open/main.php. The format of the database system allows for comprehensive molecular, clinical and epidemiological data to be deposited into a functional database, to search and manipulate the stored data and to extract and visualize the information on epidemiological, virological, clinical, nucleotide sequence and mutational aspects of HBV infection through web front-end. Specific tools, built into the database, can be utilized to analyse deposited data and provide information on HBV genotype, identify mutations with known clinical significance (e.g. vaccine escape, precore and antiviral-resistant mutations) and carry out sequence homology searches against other deposited strains. Further mechanisms are also in place to allow specific tailored searches of the database to be undertaken.

Databases, Factual↗