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[Utilizing tabacco genomic DNA to construct nearly random peptide libraries].

We developed a novel method for constructing nearly random peptide library. Genomic DNAs extracted from tissue or cells of large genome species were digested with frequent cutter to produce short DNA fragments. These short fragments can be considered nearly random. Nearly random peptide libraries can be constructed by cloning the short fragments into appropriate expression vectors and transformation into host cells. Genomic DNA from one species can be digested with different restriction enzymes and ligated to different reading frames to produce several different libraries. In this study, we digested tobacco genomic DNA with two enzymes and cloned into three different reading frames to make totally six nearly random peptide libraries.

DNA, Plant↗

[Construction of cDNA library from human colorectal cancer cell HRT-18].

OBJECTIVE: To construct a cDNA library from human colorectal cancer cell HRT-18. METHODS: The total RNA was extracted from human CRC (colorectal cancer) cell HRT-18 and the mRNA was isolated from the total RNA by MagneSphere technique, and then the first-strand cDNA was synthesized with oligo (dT) primer containing sfi I site while the double-strand cDNA was amplified through LD-PCR (long-distance PCR) by SMART technique. The double strand cDNA was digested by sfi I ( I A & I B) restriction enzyme before cDNA size fractionation, and the double-strand cDNA fractionated was ligated into the lambdaTripIEX2 vector and then was packaged into lambda phage in vitro. RESULTS: The unamplified human CRC cell HRT-18 cDNA library consisted of 6. 5 x 10(6) independent clones, recombinants clones was more than 96%. The titer of the amplified cDNA library was 1.1 x 10(9) pfu/ml and the average insert of the recombinants was 1.1 kb. CONCLUSION: The quality of the constructed cDNA library from human CRC cell HRT-18 is excellent and it is helpful to screen CRC specific antigen.

Colorectal Neoplasms↗

[The Library of Medicine of Córdoba, witness and protagonist of knowledge from the end of the 19th Century to the beginning of 21st Century].

In the nineteenth century, was founded the Medicine's Library of the National University Faculty of Medicine, center of knowledge, new ideas and teaching. The history library sector hold the valuable inheritance that represent the collection of Teachers Doctors, Pablo Mirizzi, Temístocles Castellano, Pedro Ara, José F. Verna, Juan Martín Allende located with the books and thesis of the XIX century and beginning of XX century. In 1986 it integrates the National Health Science Information Network and also, the Latino American and the Caribbean Health Science Information Network. The health libraries are in front of a new paradigm. The technologies of information and communication require new methods for administration of the information's resources and services, to answer the society information needs in the knowledge age. The traditional and virtual library is one unit that complement sources in different supports, electronic publications and interactive networks. It is the nexus between the scientific inheritance and the society, and it's mission is: Give local and virtual information's services and management the information's resources, basis of the knowledge society, support of teaching, investigation and extension of the Medical Sciences Faculty of the Córdoba National University.

Academic Medical Centers↗

Cross-species bacterial artificial chromosome (BAC) library screening via overgo-based hybridization and BAC-contig mapping of a yield enhancement quantitative trait locus (QTL) yld1.1 in the Malaysian wild rice Oryza rufipogon.

The construction of BAC-contig physical maps is an important step towards a partial or ultimate genome sequence analysis. Here, we describe our initial efforts to apply an overgo approach to screen a BAC library of the Malaysian wild rice species, Oryza rufipogon. Overgo design is based on repetitive element masking and sequence uniqueness, and uses short probes (approximately 40 bp), making this method highly efficient and specific. Pairs of 24-bp oligos that contain an 8-bp overlap were developed from the publicly available genomic sequences of the cultivated rice, O. sativa, to generate 20 overgo probes for a 1-Mb region that encompasses a yield enhancement QTL yld1.1 in O. rufipogon. The advantages of a high similarity in melting temperature, hybridization kinetics and specific activities of overgos further enabled a pooling strategy for library screening by filter hybridization. Two pools of ten overgos each were hybridized to high-density filters representing the O. rufipogon genomic BAC library. These screening tests succeeded in providing 69 PCR-verified positive hits from a total of 23,040 BAC clones of the entire O. rufipogon library. A minimal tilling path of clones was generated to contribute to a fully covered BAC-contig map of the targeted 1-Mb region. The developed protocol for overgo design based on O. sativa sequences as a comparative genomic framework, and the pooled overgo hybridization screening technique are suitable means for high-resolution physical mapping and the identification of BAC candidates for sequencing.

Chromosomes, Artificial, Bacterial↗

[Construction of a virulence-related gene library of Streptococcus mutans by suppression subtractive hybridization].

OBJECTIVE: To construct a suppression subtractive library of virulence-related genes from c serotype Streptococcus mutans (S. mutans), and lay foundations for screening the virulent genes. METHODS: After being isolated from virulent and avirulent strain of S. mutans respectively, the intact and high-pure genomic DNA was digested with three appropriate four-base-cutting restriction endonueleases to produce fragments of optimal length. The digested DNA of the virulent strain ligated with adaptor was used as tester DNA, and that of the avirulent strain as driver DNA. Then the suppression subtractive hybridization was carried out, and the efficiency of ligation and subtraction detected respectively. The subtracted fragments were inserted into vector pCR2. 1 using T/A cloning kit, and transformed into E. coli TOP10F' competent cells. Those white colonies were selected to construct the suppression subtractive library. RESULTS: Alu I chosen from three restriction endonucleases was verified to be suitable for preparing restriction fragments from S. mutans genomic DNA. Through electrophoresis of Alu I -digested DNA, a smear ranged from 0.1 to 2.0 kb was observed. The ligation efficiency of tester DNA with adaptor was at least higher than 25 percent. The subtraction efficiency of suppression subtractive hybridization confirmed the success in enrichment of differential genes between virulent and avirulent strain of S. mutans. In the subtracted group, the appearance time of the 23S rRNA gene both in tester and driver DNA was later than that in the unsubtracted group by six cycles. It suggested that suppression subtractive hybridization happened indeed. After the subtracted fragments were cloned, 96 colonies were picked up for constructing the suppression subtractive library of virulence-related genes of S. mutans. CONCLUSION: Suppression subtractive hybridization allows rapid and easy construction of virulence-related gene library of S. mutans.

Escherichia coli↗

[Bioinformatic analysis of adenoma-normal mucosa SSH library of colon].

We established a colonic adenoma-normal mucosa suppressive subtraction hybridization (SSH) library in 1999. In this study, we wanted to explore the expression profile of all candidate genes in this library. We developed an EST pipeline which contained two in-house software packages, nucleic acid analytical software and GetUni. The nucleic acid analytical software, an integrator of the universal bioinformatics tools including phred, phd2fasta, cross_match, repeatmasker and blast2.0, can blast sequences of differential clones with the downloaded non-redundant nucleotide (NR) database. GetUni can cluster these NR sequences into Unigene via matching with the downloaded Homo Sapiens UniGene database. Sixty-two candidate genes in A-N library were obtained via the high throughput automatic gene expression bioinformatics pipeline. Gene Ontology online analysis revealed that ribosome genes and immunity-regulating genes were the two most common categories in the KEGG or Biocarta Pathway. We also detected the expression of 2 genes with highest hits, Reg4 and FAM46A, by semi-quantitative RT-PCR. Both genes were up-regulated in 10 or 9 out of 10 adenomas in comparison with the paired normal mucosa, respectively. The candidate genes in A-N library would be of great significance in disclosing the molecular mechanism underlying in colonic adenoma initiation and progression.

Adenoma↗

[An optimized method for construction of genomic library].

Construction of genomic libraries is basic and important. Because of the laboriousness and high background of traditional methods for constructing genomic libraries, we improved them by overcoming these disadvantages. Two Ear I sites were chosen as the cloning sites, which can produce variable 3-base cohesive ends. Therefore the two overhangs could be devised to prevent a match and to avoid self-ligation of vector. Genomic DNA is cleaved partially with Sau3A I and subsequently incubated with dGTP and Klenow fragment of DNA polymerase Iso the self-ligation of fragments and ligation between them are blocked. In this study, the ARS probe vector (pHBM803/Trp) based on the improved method was constructed and then we constructed the Oryza sativa genomic library separately with the traditional method and improved method and compared them. The result of experiment indicated that the improved method could optimize the quality of library.

Cloning, Molecular↗

[Comparison of methods to construct a full-length cDNA library].

The use of full-length cDNA libraries is an effective tool to obtain complete gene information in a high-efficiency, high-throughput manner, especially in organisms with huge genomes that are not amenable to whole genome sequencing. In this review, we outlined several methods of full-length cDNA library construction and compared their advantages and disadvantages based on their respective principles. Drawing on our own experience, we described the Cap-trapper method in detail, with an emphasis on its application in wheat full-length cDNA library construction as well as the determination of the ratio of full-length cDNA in a library.

Cloning, Molecular↗

Team effectiveness in academic medical libraries: a multiple case study.

OBJECTIVES: The objective of this study is to apply J. Richard Hackman's framework on team effectiveness to academic medical library settings. METHODS: The study uses a qualitative, multiple case study design, employing interviews and focus groups to examine team effectiveness in three academic medical libraries. Another site was selected as a pilot to validate the research design, field procedures, and methods to be used with the cases. In all, three interviews and twelve focus groups, with approximately seventy-five participants, were conducted at the case study libraries. FINDINGS: Hackman identified five conditions leading to team effectiveness and three outcomes dimensions that defined effectiveness. The participants in this study identified additional characteristics of effectiveness that focused on enhanced communication, leadership personality and behavior, and relationship building. The study also revealed an additional outcome dimension related to the evolution of teams. CONCLUSIONS: Introducing teams into an organization is not a trivial matter. Hackman's model of effectiveness has implications for designing successful library teams.

Academic Medical Centers↗

[Preliminary approach towards construction of peptide libraries as potential tools for diagnosis of malignant thyroid tumors].

INTRODUCTION: Cancer of thyroid gland is the most common malignancy of the endocrine system. The treatment improvement could be achieved by early diagnosis. The aim of the study was to identify cancer specific antigenes with use of peptide libraries. MATERIAL AND METHODS: The material from 6 patients with thyroid cancer (4 with papillary cancer, 1 with follicular cancer and 1 with oxyphilic tumor) were analyzed. It was performed with use of lipophylic peptide libraries by direct comparison of staining of specimens prepared from normal and malignant tissue. RESULTS: Preliminary results confirm practical value of peptide libraries in early diagnostics of thyroid cancer. CONCLUSIONS: It is important to optimize construction of peptide libraries by using different staining agents hydrolyzed by proteases.

Adenocarcinoma, Follicular↗

[Construction and preliminary application of a human natural phage antibody library with diversity].

AIM: To construct a human natural phage single-chain antibody (scFv) library with diversity. METHODS: V(H) and V(L) genes were amplified by RT-PCR and hemi-PCR from peripheral blood lymphocytes of healthy persons. The V genes were assembled to form scFv by overlap PCR and cloned into phagemid pCANTAB-5E, and then transformed into E. coli TG1 by electroporation to construct a human natural phage scFv library. The diversity and gene family of antibody gene were analysed by sequencing and the specific antibodies against various antigens were screened through bio-panning. RESULTS: A human natural phage scFv library with diversity and 2x10(8) sink size was constructed successfully. The specific human scFvs against 5 antigens were obtained by bio-panning. CONCLUSION: A human natural phage scFv library with diversity is constructed successfully and can be applied to human antibody preparation.

Antibodies↗

Construction of genomic libraries of Cryptosporidium parvum and identification of antigen-encoding genes.

Genomic libraries have been constructed from bovine C. parvum DNA in the lambda ZAP and lambda DASH vectors. Based on an estimated genome size of 2 x 10(4) kilobases (kb), each recombinant library contains greater than 10 genomic equivalents. The average recombinant size for the lambda ZAP library is 2.1 kb and for the lambda DASH library is 14 kb. We have identified genes to major antigens recognized by hyperimmune bovine antiserum. These recombinants are currently being purified and characterized. Limited DNA sequence analysis of random C. parvum clones confirms suggestions that the genome is quite AT-rich. The DNA sequence of random lambda ZAP fusion proteins has identified a potential ATPase, a structural protein and a DNA-binding protein.

Animals↗

The switch library: new service in rehabilitation librarianship.

This article discusses the role of assistive devices in patient rehabilitation programs and the problems in evaluating and assessing patients for prescribing these devices. The rehabilitation library can solve many of these problems and enhance patient rehabilitation programs by providing a circulation service for adapted microswitches and other assistive devices. The article outlines library management of such a program, including funding, acquisitions, classification, and cataloging strategies. A comprehensive library classification scheme for assistive devices is proposed, and schedules are given for microswitches and environmental controls. It is predicted that this service will rapidly expand nationally over the next decade to become an integral part of rehabilitation library patient services.

Activities of Daily Living↗

The ergonomics/human factors approach to health sciences libraries.

A review of the literature reveals scant information on the application of ergonomics to health sciences libraries. Ergonomics research has identified and validated many genuine health hazards in business offices and industrial settings. While appearing innocuous, the library environment is affected by these hazards. As sophisticated technology and machinery are introduced into libraries, the human factors must be considered. This paper examines the hazards of the library environment as identified through ergonomics research and makes recommendations for alleviating or eliminating these dangers.

Air Pollutants, Occupational↗

Assaying the quality of cDNA libraries.

Once a cDNA library has been constructed, it is very useful to be able to easily assess the quality of the library. Because actin is a ubiquitous sequence, this assessment has been accomplished by probing filter lifts and Southern blots of libraries with an actin cDNA probe. These methods provide information about the percent of actin-positive clones and the degree of completeness of cDNA clones in a library. Results of these methods are correlated with the success of finding full-length clones of interest.

Actins↗

Automating Veterans Administration libraries: I. National planning and development.

The Veterans Administration Library Network (VALNET) needed an automated library system that could interface with the agency's national automation activities. The Library Special Interest Users Group was established to oversee the selection of appropriate automated systems. The Massachusetts General Hospital Utility Multi-Programming System (MUMPS) computer language and VA FILEMAN, a database management system, are being used to create new library software, which will be in the public domain.

Computer Communication Networks↗

The National Library of Medicine: from MEDLARS to the sesquicentennial and beyond.

The two decades since the introduction of MEDLARS and the passage of the Medical Library Assistance Act have been especially eventful in the history of the National Library of Medicine. The library's collections and services have grown to keep pace with the expanding health sciences literature and the needs of health professionals. Networking has emerged as an invaluable method for disseminating biomedical information. NLM has assumed new responsibilities for information services in toxicology, pharmacology, and environmental health, and for research and development in biomedical communications. Research now being carried out by NLM has the potential for enhancing the library's archival programs and for improving information dissemination in support of health sciences research, education, and practice.

Forecasting↗

Selected list of books and journals for the small medical library.

The impact that the hospital librarian's use of management techniques and comprehension of the highly competitive health care environment can have on collection development and resulting information services in his or her library is reviewed in the introduction to this revised list of 600 books and 139 journals. The list is intended as a selection guide for the small or medium-size library in a hospital or comparable medical facility, or a core collection for a consortium of small hospital libraries. Books and journals are categorized by subject; the book list is followed by an author/editor index and the subject list of journals by an alphabetical title listing. Items suggested for initial purchase by smaller libraries are indicated by asterisks. To purchase the entire collection of books and to pay for 1987 subscriptions to all journals would require about $52,600. The cost of only the asterisked items totals approximately $21,000.

Books↗