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At least 415 records · Page 23Linked to original sources
PubMed1993

Construction of subtracted cDNA libraries enriched for cDNAs for genes expressed in the mesoderm of early Xenopus gastrulae.

We present a new method for the construction of subtracted cDNA libraries. It makes use of two directional cDNA libraries in phagemid vectors and has two advantages over traditional protocols: it can be used when a limiting amount of driver material is available and the cDNA inserts in the subtracted libraries are long enough to encode functional proteins. Using this method, we have constructed two subtracted libraries enriched for sequences expressed in the mesoderm of early Xenopus gastrulae.

Animals↗
PubMed1999

Phage library-derived human anti-TETA and anti-DOTA ScFv for pretargeting RIT.

Pretargeting techniques have promise for radioimmunotherapy (RIT) in cancer because of the potential for markedly increasing the therapeutic ratio. Human antibody fragments can be retrieved from phage libraries and used to realize this potential. The library can be used to select the genetic material required to generate molecules with binding sites for both radiochelates and tumor antigens. In this study, human anti-chelate scFvs (single-chain fragments) for two metal chelates (Cu-TETA and Y-DOTA) were selected from a large, naive human scFv library. These anti-chelate scFvs were intended to serve as one arm of bispecific pretargeting molecules and to bind radiochelates given subsequently as Cu-67-TETA or Y-90-DOTA. Phage that displayed the anti-chelate scFv were selected by absorption to antibody (Lym-1) bound Cu-TETA or Y-DOTA. Enzyme-linked immunosorbent assays (ELISA) were performed to assess the intensity and specificity of phage binding to the specific chelate. Ninety-six clones demonstrating metal chelate binding seven times greater than to Lym-1 alone were chosen for diversity analysis. BstN I restriction digests were performed on DNA from these clones. Twenty-three and 43 different DNA fingerprint patterns were identified for anti-TETA and anti-DOTA clones, respectively. DNA sequencing of 39 anti-TETA clones for 23 different BstN I fingerprint patterns revealed 22 distinct sequences. Eleven of the anti-TETA clones were selected for further study. Five hundred to 1000 microg (100 to 320 microg per liter of culture) of purified scFv was produced from each of the 11 anti-TETA clones. Preliminary studies by BIAcore demonstrated evidence of 25- to 200-nM affinities. Comparable examination of the anti-DOTA clones is in progress. This study provides evidence that human scFv against unique synthetic targets can be readily selected from a large, naive human immunoglobulin phage library. Selections against metal chelated antibodies provided a wealth of scFvs with diverse binding affinities useful for engineering molecules for pretargeting RIT.

Antibodies, Monoclonal↗
PubMed1976

Educational materials reviewed for AVLINE.

The need for a central source of information regarding the availability, suitability, and quality of educational materials has led to the development of AVLINE. AVLINE is a computer information service available through the MEDLARS system of the National Library of Medicine. The Association of American Medical Colleges is gathering information from faculty members concerning useful materials and is bringing together content specialists, instructional designers, and technical specialists to review these materials. Information on materials that are recommended is provided to the National Library of Medicine to be cataloged, indexed, and abstracted for ultimate listing in AVLINE. Twenty-eight percent of the material has not been recommended; the many reasons for low ratings are categorized under content quality, instructional design, and technical quality.

Audiovisual Aids↗
PubMed1998

Structure-property correlations in a combinatorial library of degradable biomaterials.

A combinatorial library of degradable polyarylates was prepared. These polymers are A-B-type copolymers consisting of an alternating sequence of a diphenol and a diacid. The library was prepared by copolymerizing, in all possible combinations, 14 different tyrosine-derived diphenols and eight different aliphatic diacids, resulting in 8 x 14 = 112 distinct polymers. This approach (a) increases the number of available polymeric candidate materials for medical applications, and (b) facilitates the identification of correlations between polymer structure and glass transition temperature, air-water contact angle, mechanical properties, and fibroblast proliferation. The pendent chain and backbone structures were systematically varied by (a) simple homologative variations in the number of methylene groups, (b) substitution of oxygen for methylene groups, and (c) introduction of branched and aromatic structures. The polymers contained within the library exhibited incremental variations in Tg (from 2 degrees C to 91 degrees C) and air-water contact angle (from 64 degrees to 101 degrees ). Fibroblast proliferation (in vitro, serum-containing media) ranged from approximating that measured on tissue culture polystyrene to complete absence of proliferation. Generally, decreased proliferation correlated linearly with increased surface hydrophobicity, except in those polymers derived from oxygen-containing diacids in their backbone which were uniformly good growth substrates even if their surfaces were very hydrophobic. In a selected subgroup of polymers, tensile strength of thin solvent cast films ranged from about 6 to 45 MPa, while Young's modulus (stiffness) ranged from about 0.3 to 1.7 GPa. Combinatorial biomaterial libraries such as these tyrosine-derived polyarylates permit the systematic study of material-dependent biological responses and provide the medical device designer with the option to choose a suitable material from a library of related polymers that encompasses a broad range of properties.

Animals↗
PubMed1967

The National Library of Medicine and interlibrary loan.

The National Library of Medicine was a pioneer in the development of the interlibrary loan system. In 1957, the Library instituted the policy of combining loan and photocopy which it follows to this day. There has been a steady rise in the demand for photocopies from the NLM; hence, the advantage of making fuller use of local resources before approaching NLM is stressed. The criteria the Library observes to strike a balance between the national public interest expressed in the Library's statutory mission, and private interests protected by the Copyright Act, are described; but NLM will always be obliged to provide materials difficult to obtain elsewhere. Developing programs under the Medical Library Assistance Act should have considerable impact on the local availability of medical library resources.

Information Services↗
PubMed1995

Integration of information-seeking skills and activities into a problem-based curriculum.

Recent trends in medical education include a shift from the traditional, didactic, lecture-oriented approach to a more student-driven, problem-based approach to learning. This trend provides librarians with an opportunity to develop programs to teach information-gathering skills that support and are integrated into problem-based learning (PBL). In 1992, the University of Pittsburgh School of Medicine implemented the initial phase of a curriculum revision that emphasizes PBL. Since that time, Falk Library of the Health Sciences has provided a large-scale, intensive program integrating information-seeking skills and activities into the first-year Patient-Doctor Relationship course, a sequence that initiates medical school. A multimodal approach to information seeking and sources is emphasized, utilizing print and audiovisual materials, computerized resources, and subject experts. The Falk Library program emphasizes the gathering and use of information as central to both PBL and student skills development. An informal, post-course evaluation was conducted to gauge which information resources were used and valued most by students. This article presents evaluation results, including data on the use of information sources and services, and student perceptions of the librarian's role in the PBL sessions.

Curriculum↗
PubMed1970

Treasure or white elephant?

Small libraries have problems with accepting gifts of older material. Older textbooks are probably worthless, while real rarities may belong elsewhere. The usefulness of the book to the collection must be considered. While textbooks age, monographs have a longer life, and history and biography last forever. Ephemeral pamphlets have historical value. Condition of the book also affects its value. A library should always keep publications about its own institution. Suggestions are given on whether to discard or sell, with warnings about pitfalls in selling.

Book Collecting↗
PubMed1978

Health information for patients: The hospital library's role.

Libraries today, including most hospital-based patients' libraries, are involved only peripherally in providing patient health science information. Hospital libraries should collaborate with health professionals in getting health information to patients--along with the library's more traditional roles of providing recreational reading for patients and serving the informational needs of the physician and medical staff. The library should act as the center for educational materials and programs within the hospital. Many health agencies, health educators, physicians, and librarians have been discussing the need for patient health education, but there are few effectively organized or established education centers. This paper discusses an overview of patient health education and intellectural freedom, proposes a new role for the existing hospital library in patient health education, and suggests guidelines for establishing a patient education center.

Health Education↗
PubMed2003

Subtraction of cap-trapped full-length cDNA libraries to select rare transcripts.

The normalization and subtraction of highly expressed cDNAs from relatively large tissues before cloning dramatically enhanced the gene discovery by sequencing for the mouse full-length cDNA encyclopedia, but these methods have not been suitable for limited RNA materials. To normalize and subtract full-length cDNA libraries derived from limited quantities of total RNA, here we report a method to subtract plasmid libraries excised from size-unbiased amplified lambda phage cDNA libraries that avoids heavily biasing steps such as PCR and plasmid library amplification. The proportion of full-length cDNAs and the gene discovery rate are high, and library diversity can be validated by in silico randomization.

Gene Expression Profiling↗
PubMed2005

Library as place: results of a delphi study.

OBJECTIVE: An expert consensus on the future of the library as place was developed to assist health sciences librarians in designing new library spaces. METHOD: An expert panel of health sciences librarians, building consultants, architects, and information technologists was asked to reflect on the likelihood, desirability, timing, and impact on building design of more than seventy possible changes in the use of library space. RESULTS: An expert consensus predicted that the roles librarians play and the way libraries are used will substantially change. These changes come in response to changes in technology, scholarly communication, learning environments, and the health care economy. CONCLUSIONS: How health sciences library space is used will be far less consistent by 2015, as space becomes more tailored to institutional needs. However, the manner in which health sciences libraries develop and deliver services and collections will drastically change in the next decade. Libraries will continue to exist and will provide support for knowledge management and clinical trials, provide access to digital materials, and play a host of other roles that will enable libraries to emerge as institutional change agents.

Delphi Technique↗
PubMed1985

A library skills program serving adults with mental retardation: an interdisciplinary approach.

During the past decade, there has been a thrust, both socially and legally, to integrate adults with developmental disabilities, particularly those with mental retardation, into community life. Libraries, which are resources in the community for all citizens, can play a significant role in this integration. This article describes the format and outcome of a California-based library program for mentally retarded adults. It describes the materials developed, gives the contributions made by an interdisciplinary team, and discusses occupational therapy's role in the implementation of this community program.

Adult↗
PubMed2000

miniSAGE: gene expression profiling using serial analysis of gene expression from 1 microg total RNA.

The use of serial analysis of gene expression (SAGE) to determine gene expression profiles is increasing because the technique can provide absolute transcript numbers in a digital format and identify new genes. We developed a miniSAGE technique, which uses only 1 microg total RNA and reduces the amount of the starting material by 250- to 500-fold. Unlike the other modified SAGE methods, the miniSAGE technique does not require the additional PCR amplifications. The additional PCR amplifications potentially introduce bias and compromise the quantitative aspects of the SAGE method. Three key modifications in the miniSAGE technique are: (i) using the phase lock gel (PLG, Eppendorf) to increase the recovery and the purity of DNA material after each phenol extraction step; (ii) reducing the amount of linkers in the ligation, thereby minimizing their interference with SAGE ditag amplification and increasing the SAGE ditag yield; and (iii) employing the mRNA capture kit (Boehringer Mannheim) to allow the first five steps: mRNA isolation, cDNA synthesis, enzyme cleavage of cDNA, binding of the cleaved biotin-cDNA to the streptavidin-magnetic beads, ligating linkers to the bound cDNA, and the release of cDNA tags to occur within one tube to significantly reduce the loss of material between successive steps. Two fibroblast SAGE libraries have been successfully prepared. The preliminary analysis of 3838 tags from one library demonstrated a typical fibroblast gene expression pattern. This miniSAGE technique will permit a broader application of SAGE.

Biopsy↗
PubMed1991

Direct cloning of a long restriction fragment aided with a jumping clone.

Long-range physical mapping with rare-cutting restriction enzymes (rare cutters) is an important step for structural analysis of complex genomes. Combination of two types of DNA clones bearing the rare-cutter sites, linking clones and jumping clones (Fig. 1a), facilitates the physical mapping [Poustka et al., Nature 325 (1987) 353-355]. A step followed by the physical mapping is the cloning of the large (rare-cutter-generated) restriction fragment of interest. For facilitating this step, we devised a method to directly clone a long restriction fragment without constructing the whole genomic DNA library using the jumping clone as starting material. The short DNA segments of a jumping clone, which are derived from the 5' and 3' terminal regions of the large restriction fragment, are inserted into the yeast artificial chromosome plasmid (pYAC) vector, and then converted into single strands with T7 gene 6-encoded 5'----3' exonuclease. The total genomic DNA digested with the restriction enzyme is also treated with the exonuclease to convert the terminal regions of the restriction fragments into single strands. In the resulting products, only the fragment corresponding to the jumping clone can form hybrids with the just-mentioned, single-stranded DNAs, which are connected to the pYAC, and only this fragment is cloned in yeast. We describe the protocol of this method with Escherichia coli DNA as a model experiment. Judging from the cloning efficiency, this method could be applied to cloning single-copy regions of the human genome, provided a jumping clone is available. The instability of inserts in the pYAC vector is also discussed.

Base Sequence↗
PubMed1989

Isolation and characterization of a human variable copy number tandem repeat at Xcen-p11.22.

A subclone (M27B) has been isolated from a cosmid randomly selected from a library enriched for human X-chromosomal material. The subclone is extensively single-copy sequence, but also contains three complete copies and one partial copy of a 26-bp repeat, within which exists an inverted repeat having the potential to form a cruciform loop structure. Genomic sequences in this repeat region are apparently refractory to cloning, and rearrangements occurring during this process result in deletions. M27B detects multiple X-linked restriction fragments for a wide range of enzymes including MspI and HpaII. We have assigned the locus recognized by the probe (DXS255) to Xcen-Xp11.4 by mapping with a somatic cell hybrid panel and further refined its localization to Xp11.22 by in situ hybridization. The repeat sequence that is presumed to be responsible for the hypervariability observed does not show close similarity to other variable copy number tandem repeats described.

Base Sequence↗
PubMed2006

What matters most? Predictors of student satisfaction in public health educational courses.

The development of primary care trusts, the health protection agency, and non-medical public health (PH) specialists' pathways requires a competent PH workforce to be in place. Hence, the quality of education and training of PH workers is increasingly critical. With the diversity of courses and programmes that contribute to building a competent PH workforce, few studies have examined the satisfaction of students (and reasons for it) on educational PH programmes. The present study investigated the influences that four demographic variables (gender, disability, ethnicity and age bracket) and seven education-related variables (mode of study, academic term, academic level, pre-/postregistration status, entry qualification, qualification aim and class size) have on PH students' perceptions of satisfaction with 18 aspects of their learning. After controlling for the effects of the other variables, five variables explained 32% of the reported satisfaction levels. These were class size, study mode, qualification aim, pre-/postregistration status and academic level. Part-time students on postgraduate programmes are a particularly vulnerable group who may need extra support from PH educators and trainers. For these students, smaller class sizes could be beneficial, and if larger classes are inevitable, smaller group work sessions within the classes is advocated. Equally important is the availability of library resources required for learning, and course materials, handbooks and briefs need to be clear, informative, user friendly and available early in the course. Satisfied students achieved better final grades, reflecting an increased amount of transferable knowledge. The implications of the study findings for research and practice are discussed.

Adolescent↗
PubMed2002

Molecular and functional characterization of human Dectin-1.

OBJECTIVE: Dendritic cells (DC) have several unique features that differ from other antigen-presenting cells and that enable them to initiate primary immune responses. The aim of this study was to identify and characterize genes that are differentially expressed in DC generated from CD14+ peripheral blood monocytes in vitro. MATERIALS AND METHODS: Using a subtractive cDNA library, we identified the full-length cDNA sequence of human Dectin-1b by rapid amplification of cDNA ends. Expression profiles were performed by reverse transcriptase polymerase chain reaction and Western blotting. For functional analysis, HeLa cells were transfected with the Dectin-1b coding region and used as stimulators for purified peripheral blood T-cell populations. RESULTS: Our subtractive cloning strategy revealed the human Dectin-1b cDNA, which encodes for a transmembrane protein of the C-type lectin-like receptor family. It is selectively expressed in several purified DC subpopulations but not in monocytes and is up-regulated upon stimulation with lipopolysaccharide. The nucleotide sequence was submitted to GenBank (accession no. AY009090). Furthermore, we demonstrate that the Dectin-1b splice variant transfected in HeLa cells up-regulates the activation markers on human T lymphocytes, induces the production of interferon-gamma, and promotes proliferation of both CD4+ and CD8+ T lymphocytes. CONCLUSION: The Dectin-1 gene is expressed during the development of DC from peripheral blood monocytes and the transfection of the splice variant 1b into HeLa cells results in the stimulation of effector functions of human T lymphocytes.

Antigens, CD↗
PubMed2003

Combined modality radiotherapy and chemotherapy in nonsurgical management of localized carcinoma of the esophagus: a practice guideline.

PURPOSE: To make recommendations regarding combined radiotherapy (RT) and chemotherapy (RTCT), compared with RT alone, when a nonsurgical approach is used for patients with localized esophageal carcinoma. MATERIALS AND METHODS: The Medline, Cancerlit, Cochrane Library databases, and abstracts published in the American Society of Clinical Oncology and the American Society for Therapeutic Radiology and Oncology proceedings were searched for evidence. Evidence was evaluated by two members of the Gastrointestinal Cancer Disease Site Group and methodologists. RESULTS: Pooling seven randomized trials detected a statistically significant survival benefit at 1 year for concomitant RTCT compared with RT alone (1-year mortality odds ratio 0.61; 95% confidence interval 0.44-0.84; p <0.00001). Local control also significantly improved with concomitant RTCT compared with RT alone for the available data (odds ratio 0.52; 95% confidence interval 0.31-0.89; p = 0.004), but a significant increase in adverse effects, including life-threatening toxicities, was shown. CONCLUSIONS: Concomitant RT and cisplatin-based CT is recommended over RT alone. Patients should be aware of the increased acute toxicity associated with this approach, and this recommendation should only be made after consideration of the potential risks and benefits and the patient's general condition. Sequential RTCT is not recommended as standard practice.

Adenocarcinoma↗