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Filling the gaps in the porcine linkage map: isolation of microsatellites from chromosome 18 using flow sorting and SINE-PCR.

Flow-sorted chromosome 18 material from the pig (SSC18) was amplified with SINE-PCR to generate DNA for molecular cloning. An SSC18-enriched library was constructed and subsequently screened with a (CA)15 probe to identify polymorphic microsatellites. Eleven unique microsatellites were obtained, six of which constituted direct extensions of SINE poly(A) tracts. Eight primer pairs amplified polymorphic loci in unrelated pigs. The polymorphic markers were typed in a Swedish reference pedigree for porcine genome mapping, but only two of them (S0177 and S0179) mapped to SSC18. The remaining markers mapped to chromosomes 3, 4, 7, 9, and 11. Together with two markers from other sources (S0062 and sw787), a four-point linkage group on SSC18 could be established.

Animals↗

The asthma year in Swedish pharmacies: a nationwide information and pharmaceutical care program for patients with asthma.

OBJECTIVE: To increase the understanding of asthma among patients and the general public in Sweden and to increase the collaboration among healthcare personnel for the benefit of asthma patients. DESIGN: Based on the results from problem detection studies, Apoteksbolaget, the national organization for pharmacies in Sweden, designed a program to educate healthcare professionals and the public about asthma. Entitled the "Asthma Year," all pharmacy staffs were offered education on asthma, patients were provided written information and audiovisual materials, and the general public was made more aware of asthma through local media. SETTING: Pharmacies, healthcare centers, hospital outpatient areas, libraries, places of work, and domestic airports. PARTICIPANTS: All pharmacy staffs (12,000 persons) were offered an education on asthma. Patients with asthma were offered brochures and leaflets in pharmacies and information in the pharmacy magazine Apoteket. The general public received information about asthma through posters, radio, and television. MAIN OUTCOME MEASURES: The evaluations of the Asthma Year indicate that the activities had an impact on the public, on patients with asthma, and on the pharmacy staffs. RESULTS: About 4.5 million people in Sweden (42% of the population) observed the Asthma Year or received information about asthma, and 49% of the patients learned something new about their disease. Twice as many patients used steroid therapy regularly after the campaign than before. Of the pharmacy staffs, 96% were favorable to the idea of working with campaign years and 85% believed that they were personally involved with the activities. CONCLUSIONS: Patient outcomes in terms of reduced morbidity have not been possible to document, but the number of deaths from asthma in Sweden decreased from 438 during the previous year to 398 during the Asthma Year.

Adolescent↗

Ras-association domain family 1 protein, RASSF1C, is an IGFBP-5 binding partner and a potential regulator of osteoblast cell proliferation.

UNLABELLED: The goal of this study was to identify downstream signaling molecules involved in mediating the IGF-independent effects of IGFBP-5 in osteoblasts. We identified RASSF1C, a member of the RASSF1 gene products, as a IGFBP-5 binding partner and as a potential mediator of IGFBP-5 effects on ERK phosphorylation and cell proliferation. INTRODUCTION: It has been predicted that the intrinsic growth factor action of insulin-like growth factor binding protein (IGFBP)-5 involves either the binding of IGFBP-5 to a putative receptor to induce downstream signaling pathways and/or intracellular translocation of IGFBP-5 to bind to potential signaling molecules involved in osteoblast cell regulation. This study reports the characterization of isoform C of the Ras association family 1 (RASSF1C) gene as an interacting partner of IGFBP-5. MATERIALS AND METHODS: IGFBP-5 was used as bait in a yeast two-hybrid screen of a human osteosarcoma cDNA library. Expression levels of RASSF1C were measured by RT-PCR and/or Northern blot. IGFBP-5 effects on ERK phosphorylation were evaluated by immunoblot analysis. The effect of RASSF1C siRNA on cell proliferation was measured by the AlamarBlue assay. RESULTS: One of the clones that interacted strongly with the bait under high stringency conditions corresponded to RASSF1C. The interaction between RASSF1C and IGFBP-5 was confirmed by in vitro co-immunoprecipitation studies. Northern blot and RT-PCR analysis showed that RASSF1C was expressed in a variety of osteoblast cell types that produce IGFBP-5. Addition of synthetic RASSF1C-specific small interfering (si) RNA duplex or use of a RASSF1C-specific si-hairpin plasmid caused a decrease in cell number and abolished IGFBP-5-induced extracellular signal-regulated kinase (ERK)-1/2 phosphorylation but had no effect on IGFBP-5-induced increases in alkaline phosphatase (ALP) activity. CONCLUSIONS: We have shown a novel interaction between IGFBP-5 and RASSF1C. Our findings that silencing of RASSF1C results in the reduction of osteoblast cell proliferation and that IGFBP-5 treatment increases phosphorylation of ERK-1/2 raise the possibility that RASSF1C, a Ras effector, could, in part, contribute to mediating the effects of IGFBP-5 on ERK phosphorylation and, consequently, cell proliferation.

Amino Acid Sequence↗

Faxon Finder/Faxon Xpress: report from a beta test site.

In this environment of expanding information needs, spiraling journal costs, and curtailed financial resources, medical libraries must take advantage of the rapid evolution in document delivery services. There are exciting developments in transmission technology, from fax machines to scanners to the Internet. There is also a notable shift away from the traditional reliance on library networks toward use of commercial vendors as document providers. These changes require reevaluation of interlibrary loan (ILL) and document delivery systems on many levels, including pricing structures, work flow, and impact on collection development. As commercial vendors develop increasingly effective products, librarians can test, evaluate, and incorporate them into their ILL operations to enhance service to users and relieve the pressure to acquire new resources; in this way, librarians can move from the "just in case" practice of building an on-site collection to the "just in time" model of providing timely delivery of materials to users as needed. This paper describes a beta test of the Faxon Finder/Faxon Xpress table of contents and document delivery services conducted by the Treadwell Library of the Massachusetts General Hospital.

Databases, Bibliographic↗

1927 reference in the new millennium: where is the Automat?

James Ballard, director at the Boston Medical Library, tracked questions he received at the reference desk in 1927 to recognize the trend of queries and to record the information for future use. He presented a paper on reference services that listed sixty of his reference questions at the Thirtieth Annual Meeting of the Medical Library Association (MLA) in 1927. During a two-month period in 2001, the authors examined Ballard's questions by attempting to answer them with print sources from the 1920s and with the Internet. The searchers answered 85% of the questions with the Internet and 80% with 1920s reference sources. The authors compared Internet and 1920s print resources for practical use. When answering the questions with 1920s resources, the searchers rediscovered a time in health sciences libraries when there was no Ulrich's Periodicals Directory, no standardized subject headings, and no comprehensive listings of available books. Yet, the authors found many of the 1920s reference materials to be quite useful and often multifunctional. The authors recorded observations regarding the impact of automation on answering reference questions. Even though the Internet has changed the outward appearance of reference services, many things remain the same.

History, 20th Century↗

"AND GLADLY TEACH": THE AMERICAN HOSPITAL ASSOCIATION'S EXPERIENCE IN CONDUCTING INSTITUTES ON HOSPITAL LIBRARIANSHIP.

As part of its overall educational program, the American Hospital Association has since 1959 conducted three institutes on hospital librarianship to meet the demand for more competent librarians in medical, nursing school, and patients' libraries. The purpose of such institutes is to teach the basic elements of library science to untrained personnel in hospital libraries. Discussed are steps in initiating an institute; factors determining length, date, and place; financing; publicity; choice and responsibility of local advisory committee; program content; qualifications of instructors; characteristics of registrants; materials for distribution; evaluations. Details of the most recent institute are outlined. A summary of problems still facing this type of educational program and suggestions for future improvements conclude the paper.

Academies and Institutes↗

The role of National Library of Medicine web sites in newborn screening education.

Expanded newborn screening programs and subsequent detection of rare genetic disorders challenge parents and their medical providers to learn about the treatment and management of these disorders. Many people seek medical information on the Internet but may encounter requests for registration or fees, or find that resources are out of date, difficult to understand, or buried in advertisements. The U.S. National Library of Medicine (NLM), a component of the National Institutes of Health, provides web-based resources that address the challenges of newborn screening education. These resources include MedlinePlus, Genetics Home Reference, ClinicalTrials.gov, and PubMed. NLM websites are not commercial, do not require registration or fees, and provide varied levels of information for a continuum of audiences from low-literacy consumers to health professionals. Using phenylketonuria as an example, this study describes the information that parents and their medical providers can find through NLM resources. NLM has embraced the digital age and provides the public with reliable, accurate, and up-to-date educational materials.

Genetic Testing↗

The construction of Streptomyces cyaneus genomic libraries in Escherichia coli is dependent upon the use of Mcr-deficient strains.

Streptomyces cyaneus genomic DNA ligated into either lambda phage or plasmid vectors was very inefficiently cloned into standard Escherichia coli host strains. However, the same material could be efficiently cloned using Mcr-deficient E. coli strains. These results suggest that the S. cyaneus genome contains 5-methylcytosine residues, some of which occur within the recognition sequences of the E. coli Mcr restriction system.

Cloning, Molecular↗

DNA display II. Genetic manipulation of combinatorial chemistry libraries for small-molecule evolution.

Biological in vitro selection techniques, such as RNA aptamer methods and mRNA display, have proven to be powerful approaches for engineering molecules with novel functions. These techniques are based on iterative amplification of biopolymer libraries, interposed by selection for a desired functional property. Rare, promising compounds are enriched over multiple generations of a constantly replicating molecular population, and subsequently identified. The restriction of such methods to DNA, RNA, and polypeptides precludes their use for small-molecule discovery. To overcome this limitation, we have directed the synthesis of combinatorial chemistry libraries with DNA "genes," making possible iterative amplification of a nonbiological molecular species. By differential hybridization during the course of a traditional split-and-pool combinatorial synthesis, the DNA sequence of each gene is read out and translated into a unique small-molecule structure. This "chemical translation" provides practical access to synthetic compound populations 1 million-fold more complex than state-of-the-art combinatorial libraries. We carried out an in vitro selection experiment (iterated chemical translation, selection, and amplification) on a library of 10(6) nonnatural peptides. The library converged over three generations to a high-affinity protein ligand. The ability to genetically encode diverse classes of synthetic transformations enables the in vitro selection and potential evolution of an essentially limitless collection of compound families, opening new avenues to drug discovery, catalyst design, and the development of a materials science "biology."

Catalysis↗

Construction of a large phage display antibody library by in vitro package and in vivo recombination.

Capacity and diversity are extremely important to the quality of various phage display libraries. In this work, lambda phage-based in vitro package was applied to construct a filamentous phage display antibody library so as to enlarge its capacity and introduce more sequence diversity in the final library. In vivo recombination via Cre recombinase/lox sites was also exploited to create V(H)/V(L) combination diversity based on multivalent package of lambda phage packaging extracts on phagemid DNA concatemers. The library constructed with 10 microg concatenated phagemid DNA and ten vials of lambda phage packaging extracts was calculated to contain 1.40 x 10(10) independent clones. Higher capacity can be easily achieved when more materials are consumed. This strategy is somewhat more efficient than prior methods.

Antibodies↗

Drug information libraries on the Internet.

UNLABELLED: In the past several years there has been a dramatic proliferation of drug-related sites on the Internet. This article reviews the information found at selected Internet drug information libraries, and comments on its accuracy and implications. Drug-related sites were found by initially performing an Internet search on "psychoactive drugs" and then exploring links among the sites identified. Sites were chosen on the basis of comprehensiveness of information and positive or tolerant attitude toward drug use. While all classes of drugs are discussed at these sites, the primary foci of interest are synthetic and naturally occurring hallucinogens. Many of the biological materials discussed are legal and readily available. Information surveyed at these sites was largely accurate regarding the effects of various substances and biological sources of psychoactive compounds. CONCLUSIONS: Internet drug information libraries contain large amounts of information about a wide variety of drugs, including previously little-known biological sources of hallucinogens. The availability of this information could have significant effects on patterns of drug use.

Information Services↗

Rapid verification of identity and content of drug formulations using mid-infrared spectroscopy.

A general method for the rapid verification of both identity and content of complete solid drug formulations has been devised. Infrared spectra for the samples were recorded using the diffuse reflectance technique, and specially written software was employed to identify the type of formulation and level of active ingredient. This software was devised to ensure reliable use when applied by those with minimal operator skills. Three differing drug tablet formulations containing simvastatin, enalapril maleate and lovastatin, as well as a capsule formulation containing finastride were studied. Adequate precision was obtained to reliably verify drug dosage levels. Near-infrared (NIR) and mid-infrared (MIR) spectrometers were evaluated for use with the method. The MIR instrument allowed sufficient resolution and spectral/structural selectivity to reliably verify correctness of either of two near derivative drugs necessarily present in the same clinical study. Drug tablet and capsule dosage levels tested ranged from 0.2 to 40 mg of drug. Approximately 1% (w/w) of the drug in the formulation was the minimum amount determined. Parameters affecting method ruggedness in routine use were optimized. Experimental addition of an extraneous material to a simvastatin formulation was easily detected and flagged by the routine test procedure. Subsequent data retrieval and searching against spectral libraries was used to demonstrate identification of the additive.

Anticholesteremic Agents↗

Quality assurance of treatment plans for interstitial and intracavitary high-dose-rate brachytherapy.

PURPOSE: Quality assurance for complex high-dose-rate (HDR) treatment planning has always been a challenge to the physics community because of the time constraint between HDR planning and the delivery of the treatment. This study proposes an efficient, precise, and easy method for checking the complex computer calculation. METHODS AND MATERIALS: Posttreatment, three-dimensional dose-volume study was performed for 98 patients with 128 new treatment plans along with 30 library plans. Volumes covered by the 100% isodose line, source activity (Ci), total dwell time (s), and the prescription dose (100%) were recorded. Variation of R(V) defined as (irradiated time x activity/elongation factor x prescribed dose) with volume was studied for different catheter systems. RESULTS: Parametric fit of R(V) with volume for three different systems that cover most of the interstitial and intracavitary brachytherapy implants agrees within +/-6%. CONCLUSIONS: The excellent agreement of R(V) derived from this simplistic point source model with three-dimensional dose calculations for individual HDR treatment plans clearly establishes that for an implant with known number of catheters, the time needed to deliver a prescribed dose to a given prescription volume can be easily predicted.

Brachytherapy↗

Molecular analysis of the hematopoiesis supporting osteoblastic cell line U2-OS.

OBJECTIVE: Osteoblasts play an important role in regulating hematopoiesis in the bone marrow. Here we show that U2-OS, a widely used osteoblastic cell line derived from an osteosarcoma, has the capacity to support proliferation of human hematopoietic progenitor cells in vitro. In this study, U2-OS cells are characterized at the molecular level to unravel the molecular mechanisms underlying the support of hematopoiesis. MATERIALS AND METHODS: U2-OS was analyzed in great detail using RT-PCR and flow cytometry. In addition, a cDNA library was constructed and randomly sequenced to obtain insight in the repertoire of expressed molecules. RESULTS: A broad panel of growth factors and cytokines is expressed by U2-OS. TGF-beta, GM-CSF, c-kit ligand, and IL-7 are produced constitutively and IL-1beta, IL-6, IL-8, TNF-alpha, IFN-gamma, and MIP1-alpha are upregulated upon stimulation. In addition to those, mRNAs of the CC chemokine LARC and leukemia inhibitory factor were identified. U2-OS cells express high levels of beta1-integrins at the cell surface: VLA-2, VLA-3, VLA-4, VLA-5, VLA-6, and the integrin alphavbeta3. Besides integrins, ALCAM and NCAM are detected on the cell surface of U2-OS. Interestingly, we show that CD34(+) progenitor cells expressing ALCAM are highly proliferative when compared with CD34(+) ALCAM(low) cells, hinting at a role for ALCAM in anchoring progenitor cells to the bone marrow stroma. Interestingly, random sequencing of an U2-OS cDNA library yielded almost 10% of novel cDNAs with a potential role in hematopoiesis. The involvement of these novel molecules in hematopoiesis is an interesting target for future investigations. CONCLUSIONS: We conclude that U2-OS supports outgrowth of hematopoietic progenitor cells and accordingly expresses adhesion molecules and growth factors and a number of novel, as yet uncharacterized potentially interesting genes.

Activated-Leukocyte Cell Adhesion Molecule↗

Analysis of the conserved N-terminal domains in major ampullate spider silk proteins.

Major ampullate silk, also known as dragline silk, is one of the strongest biomaterials known. This silk is composed of two proteins, major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2). Only partial cDNA sequences have been obtained for these proteins, and these sequences are toward the C-terminus. Thus, the N-terminal domains have never been characterized for either protein. Here we report the sequence of the N-terminal region of major ampullate silk proteins from three spider species: Argiope trifasciata, Latrodectus geometricus, and Nephila inaurata madagascariensis. The amino acid sequences are inferred from genomic DNA clones. Northern blotting experiments suggest that the predicted 5' end of the transcripts are present in fibroin mRNA. The presence of more than one Met codon in the N-terminal region indicates the possibility of translation of both a long and a short isoform. The size of the short isoform is consistent with the published, cDNA based, N-terminal sequence found in flagelliform silk. Analyses comparing the level of identity of all known spider silk N-termini show that the N-terminus is the most conserved part of silk proteins. Two DNA sequence motifs identified upstream of the putative transcription start site are potential silk fibroin promoter elements.

Amino Acid Sequence↗

The Nuclear Receptor Resource: a growing family.

Last year, the original Glucocorticoid Receptor Resource was expanded into a comprehensive project: the Nuclear Receptor Resource (NRR, http:// nrr.georgetown.edu/nrr/nrr.html ). The NRR has since been offering comprehensive information on nuclear receptor structure and function, as well as general facts of interest to the scientific community on meetings, funding and employment opportunities. The project now includes individual resources as part of a network which integrates information on glucocorticoid, androgen, mineralocorticoid, thyroid hormone, Vitamin D and peroxisome-proliferator activated receptors. Many investigators have joined the NRR network by filling the Who is who? form available in the NRR home page. This has facilitated communication among scientists in the field and dissemination of data nor otherwise published. Because several investigators have contacted NRR authors over the past few months asking for advice and materials for educational purposes, we have recently decided to include in our project an educational resource on nuclear receptors termed the 'Graphics Library'. The input and suggestions of NRR users do shape the future direction of the project, so we encourage user to give us feedback.

Computer Communication Networks↗

SOURCES: a code for calculating (alpha,n), spontaneous fission, and delayed neutron sources and spectra.

SOURCES is a computer code that determines neutron production rates and spectra from (alpha,n) reactions, spontaneous fission and delayed neutron emission owing to the decay of radionuclides in homogeneous media, interface problems and three-region interface problems. The code is also capable of calculating the neutron production rates due to (alpha,n) reactions induced by a monoenergetic beam of alpha particles incident on a slab of target material. The (alpha,n) spectra are calculated using an assumed isotropic angular distribution in the centre-of-mass system with a library of 107 nuclide decay alpha-particle spectra, 24 sets of measured and/or evaluated (alpha,n) cross sections and product nuclide level branching fractions, and functional alpha particle stopping cross sections for Z < 106. Spontaneous fission sources and spectra are calculated with evaluated half-life, spontaneous fission branching and Watt spectrum parameters for 44 actinides. The delayed neutron spectra are taken from an evaluated library of 105 precursors. The code outputs the magnitude and spectra of the resultant neutron sources. It also provides an analysis of the contributions to that source by each nuclide in the problem.

Algorithms↗

Management of urinary tract infections: historical perspective and current strategies: Part 1--Before antibiotics.

PURPOSE: Urinary tract infections (UTIs) have plagued mankind long before bacteria were recognized as the causative agents of disease and before urology became an established medical specialty. To our knowledge a comprehensive review of the recorded medical history of UTI from its first description in ancient Egyptian papyri through today has not been attempted until now. MATERIALS AND METHODS: Numerous resources were used to collect the information described in this review. Older textbooks from the school of medicine library at our institution were used to collect information on UTI in ancient times. Medical texts from the 19th century contains information regarding the treatment of UTI during that era. Early volumes of the Journal of Urology from the beginning of the 20th century are rich with information on the first attempts at chemotherapy for UTI. MEDLINE searches were used to collect appropriate information after 1969. RESULTS: The Ebers papyrus from ancient Egypt recommended herbal treatments to ameliorate urinary symptoms without providing insight into pathological mechanisms. Hippocrates believed that disease was caused by disharmony of the 4 humors and accordingly diagnosed urinary disorders. Roman medicine further expanded the conservative approach (bed rest, diet, narcotics and herbs) advocated by Greek physicians, while also improving invasive techniques (surgical lithotomy for stones and catheterization for retention). The Arabian physician Aetius refined uroscopy and created a detailed classification and interpretation of urinary disease based on this technique. During the Middle Ages no major advances occurred, although existing therapies were refined and treatments for gonococcal urethritis were well described. The early 19th century provided vivid and detailed descriptions of UTIs without the knowledge that they were caused by microorganisms. Management included hospitalization, bed rest, attention to diet, plasters, narcotics, herbal enemas and douches, judicious bleeding (direct bleeding, cupping and leeches), and surgery for stones, abscess and retention. The discovery of microorganisms as the etiological agents of infectious diseases in general and inflammation associated with urinary diseases in particular provided an impetus for physicians critically to examine management approaches and develop evidence based strategies for UTI treatment. Various antibacterial agents, such as hexamine, mercurochrome and others (hexylresorcinol, methylene blue, pyridium, acriflavin and mandelic acid), showed promise in laboratory studies but their efficacy in clinical investigations was disappointing. CONCLUSIONS: Treatments for UTI until the discovery of antibiotics were largely palliative because the bacterial origin of UTI was not recognized and no specific antimicrobial therapies were available.

China↗