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Isolation of a human homolog of osteoclast inhibitory lectin that inhibits the formation and function of osteoclasts.

UNLABELLED: Osteoclast inhibitory lectin (OCIL) is a newly recognized inhibitor of osteoclast formation. We identified a human homolog of OCIL and its gene, determined its regulation in human osteoblast cell lines, and established that it can inhibit murine and human osteoclast formation and resorption. OCIL shows promise as a new antiresorptive. INTRODUCTION: Murine and rat osteoclast inhibitory lectins (mOCIL and rOCIL, respectively) are type II membrane C-type lectins expressed by osteoblasts and other extraskeletal tissues, with the extracellular domain of each, expressed as a recombinant protein, able to inhibit in vitro osteoclast formation. MATERIALS AND METHODS: We isolated the human homolog of OCIL (hOCIL) from a human fetal cDNA library that predicts a 191 amino acid type II membrane protein, with the 112 amino acid C-type lectin region in the extracellular domain having 53% identity with the C-type lectin sequences of rOCIL and mOCIL. The extracellular domain of hOCIL was expressed as a soluble recombinant protein in E. coli, and its biological effects were determined. RESULTS AND CONCLUSIONS: The hOCIL gene is 25 kb in length, comprised of five exons, and is a member of a superfamily of natural killer (NK) cell receptors encoded by the NK gene complex located on chromosome 12. Human OCIL mRNA expression is upregulated by interleukin (IL)-1alpha and prostaglandin E2 (PGE2) in a time-dependent manner in human osteogenic sarcoma MG63 cells, but not by dexamethasone or 1,25 dihydroxyvitamin D3. Soluble recombinant hOCIL had biological effects comparable with recombinant mOCIL on human and murine osteoclastogenesis. In addition to its capacity to limit osteoclast formation, OCIL was also able to inhibit bone resorption by mature, giant-cell tumor-derived osteoclasts. Thus, a human homolog of OCIL exists that is highly conserved with mOCIL in its primary amino acid sequence (C-lectin domain), genomic structure, and activity to inhibit osteoclastogenesis.

Acid Phosphatase↗

[From temples and sanctuaries to hospitals; 6,000 years of history].

OBJECTIVE: To find out when and where the hospitals began, its history and evolution. MATERIAL: Review of books and Journals of personal and Instituto Nacional de la Nutrición Salvador Zubirán, libraries about history of medicine and surgery. RESULTS: During the last 5000 years the history of man has been full of events, the beginning and development of hospitals has been one of them. Everything started in Sumeria in 3500 bC, some changes were developed in Egypt, Greece and Rome. In the 4th century aD the first true hospitals are opened. Since then the hospitals had improved until now. CONCLUSIONS: The sick man has been looking for the healing of his ailments, and few places made him feel protected as the hospital does. The birth, growing and evolution of such institutions had been slow and steady until the end of the second millennium aD.

History, 15th Century↗

[The Lwow Medical Chamber].

The materials related to the Lwow Medical Chamber comprised in the Special Collection of the Main Medical Library date back to the establishment of the Medical Chambers. They cover the period from 1893 to 1945 and provide information on the details of the operating area of the Lwow Medical Chamber, the number of its members, functions of individual presidents and the composition of changing managements and boards. They also include information on the legal acts regulating the operations of the Medical Chambers and their scope of activities. The archive files incorporated in the Special Collection of the Main Medical Library include 216 archive units, which represent only a small percentage of preserved files: physicians' personal files including personal questionnaires, registration cards and photographs. The articles discuss the above issues extensively.

Archives↗

Overexpression of human phosphoglycerate kinase 1 (PGK1) induces a multidrug resistance phenotype.

BACKGROUND: Multidrug resistance is a significant barrier to the development of successful cancer treatment. To identify genetic alterations that are directly involved in paclitaxel resistance, a functional cloning strategy was developed. MATERIALS AND METHODS: Using mRNA from paclitaxel resistant human ovarian cancer cell line SW626TR, a cDNA library was established in a pCMV-Script vector that permits expression of cDNA inserts in mammalian cells. Transfection of the pCMV-Script/SW626TR cDNA library into the paclitaxel-sensitive human osteogenic sarcoma cell line, U-20S, resulted in several paclitaxel-resistant clones. RESULTS: DNA sequencing of clone C16 demonstrates complete homology to human phosphoglycerate kinase 1 (PGK1). Retransfection of the PGK1 insert into U-20S confers a multidrug resistant phenotype, characterized by a 30-fold increase in paclitaxel resistance, and cross-resistance to vincristine; adriamycin and mitoxantrone, but not methotrexate or cisplatin. Enzymatic analysis of the PGK1 transfectants demonstrates an increase in PGK1 activity as compared to the parental cell line, U-20S. Northern and Western analysis of PGK1 transfectants reveals no change in MDR-1 expression compared with the parental cell line. In addition, co-culture of PGK1 transfectants with verapamil only partially reverses the multidrug resistant phenotype. Rhodamine 123 studies are also consistent with an MDR-1 independent mechanism of increased drug efflux. CONCLUSION: Together this data suggests that PGK1 can induce a multidrug resistant phenotype through an MDR-1 independent mechanism.

Antineoplastic Agents↗

[A new method for screening mutant yeast strains with green fluorescent protein].

OBJECTIVE: To establish a fast and simple method for screening mutant yeast strains. MATERIALS AND METHODS: Homologous recombination technique was used to detect mutant yeast strains in yeast genomic library, with the green fluorescent protein gene as the reporter gene in the transposon. RESULTS: The strains that emitted green fluorescence were isolated, indicating that the gfp gene was inserted into the yeast genome by homologous recombination. CONCLUSION: This study established a useful method for functional genome study by homologous recombination technique, and provide an alternative for gene therapeutic drug development.

Cloning, Molecular↗

Interstitial deletion of chromosome 9q with coexistence of the deleted segment as a ring chromosome. A case report.

In a mentally retarded female an interstitial deletion of a chromosome 9 and an additional ring chromosome was shown, which by positive hybridisation with a no 9 library was considered to be the excised segment. The functional centromere and C and DA/DAPI positive material as well on the ring chromosome are explained by a break within the centromere close to the constitutive heterochromatin and supports the hypothesis of "latent" centromere(s).

Chromosome Deletion↗

Health promotion and prevention.

Techniques of promotive and preventive health care include: The use of proactive skills in the consultation; and the co-ordinated use of posters and pamphlets with media campaigns. An efficient medical record and patient recall system; and a doctor initiated health check programme with selected patients. A cost effective method of informing practitioners includes: Ensuring access to continuing medical education by supporting FMP and post-graduate medical institutes. The provision of educational material for both the patient and doctor combined with media publicity campaigns. Supporting RACGP CHECKUP and library services for isolated practitioners. Suggested changes to undergraduate medical education are: Promotive and preventive health care delivery needs to be an inbuilt part of the curriculum. Promotive and preventive health care needs to be taught by both academic and private general practitioners who have received tutor training. It should be presented in tutorial form and during student attachment at surgeries. It needs to be followed up by continuing education by postgraduate institutes, the RACGP and FMP. Government input required includes: The provision of increased academic resources and continued funding of FMP. Remuneration for general practitioners who are involved in undergraduate teaching. Alteration of the fee structure to reward quality of care. Allowing Medicare rebate for promotive and preventive health care delivery.

Health Promotion↗

The Modular Resource Center: integrated units for the study of the anatomical sciences in a problem-based curriculum.

The Modular Resource Center (MRC) at the College of Veterinary Medicine at Cornell University was created in 1993 as a way to provide visual resources in support of a newly implemented problem-based curriculum in which the anatomical sciences are taught primarily in the first tutorial-based course, The Animal Body. Over two dozen modules have been created specifically in support of this course, whereas additional modules have been created in support of other basic science courses. The basic unit of organization of the MRC is a module presented in a carrel that provides students a way to study, either alone or in groups, a given topic. The topic is presented through a script and an integrated set of anatomical materials including plastinated dissected specimens, vascular casts, skeletal preparations, models, radiographs, histological slides, and photo- and electron micrographs. The key feature of this resource center is that it is not a museum; rather it is more analogous to an interactive library, that can be used for reference, study, and review, not only by veterinary students but also by faculty, interns, residents, and undergraduates. A unique aspect is that all materials have been made by veterinary students working with faculty during the summer. Although started as a resource in support of a tutorial-based curriculum, the MRC has evolved over a decade into an anatomy resource that would be highly valued in any curricular format.

Anatomy↗

Novel method for high throughput DNA methylation marker evaluation using PNA-probe library hybridization and MALDI-TOF detection.

The methylation of CpG dinucleotides has become a topic of great interest in cancer research, and the methylation of promoter regions of several tumor suppressor genes has been identified as a marker of tumorigenesis. Evaluation of DNA methylation markers in tumor tissue requires hundreds of samples, which must be analyzed quantitatively due to the heterogeneous composition of biological material. Therefore novel, fast and inexpensive methods for high throughput analysis are needed. Here we introduce a new assay based on peptide nucleic acid (PNA)-library hybridization and subsequent MALDI-TOF analysis. This method is multiplexable, allows the use of standard 384 well automated pipetting, and is more specific and flexible than established methods, such as microarrays and MS-SNuPE. The approach was used to evaluate three candidate colon cancer methylation markers previously identified in a microarray study. The methylation of the genes Ade-nomatous polyposis coli (APC), glycogen synthase kinase-beta-3 (GSK3beta) and eyes absent 4 (EYA4) was analyzed in 12 colon cancer and 12 normal tissues. APC and EYA4 were confirmed as being differentially methylated in colon cancer patients whereas GSK3beta did not show differential methylation.

Colorectal Neoplasms↗

Chromosome in situ suppression hybridisation in clinical cytogenetics.

The use of chromosome in situ suppression hybridisation with whole chromosome libraries has previously been reported by various research laboratories to be an effective method of identifying specific human chromosomal material. As a clinical cytogenetic service laboratory we have used the technique as a complement to diagnosis by classical chromosome banding. In three examples of structural rearrangements the potential use of the 'chromosome painting' method is assessed for its ability to enhance the routine cytogenetic service currently available.

Adult↗

Sorting a Staphylococcus aureus phage display library against ex vivo biomaterial.

A phage display library made from Staphylococcus aureus DNA was sorted against a central venous catheter (CVC) that had been removed from a patient 2 days after insertion. After the first panning, approximately 50% of the clones encoded proteins known to interact with mammalian proteins. After the second and third pannings, fibrinogen-binding and beta2-glycoprotein I (beta2-GPI)-binding phage particles were clearly dominating. Proteins adsorbed to different CVCs were investigated using specific antibodies. Among the proteins probed for, fibrinogen was most abundant, but, interestingly, beta2-GPI was also detected on all tested CVCs.

Bacterial Adhesion↗

Seniors connect: a partnership for training between health care and public libraries.

Kaleida Health Libraries, in partnership with the Buffalo and Erie County Public Library, presented "Seniors Connect: A Health Information Project." The program was funded by the Community Foundation for Greater Buffalo and the Kaleida Health Foundation. The purpose of the project was to teach African-American and Hispanic senior citizens or their caregivers residing in the city of Buffalo how to search the Internet to find quality, accurate, understandable health information. Attendees were taught to distinguish quality information from inaccurate, misleading, and fraudulent material. A total of fourteen training sessions were offered. The Seniors Connect program was initiated with health fair at each branch library, conducted by Kaleida Health staff, to provide an opportunity for participants to learn about health care services available in their community; observe demonstrations of health information searches on the Internet; learn about new treatment modalities for diseases such as diabetes, high cholesterol, thyroid abnormalities, and hypertension; and interact with area health care providers and library staff. The sessions were highly rated by attendees and many participants expressed their gratitude for a program geared specifically toward seniors.

Black or African American↗

On-line FT-Raman and dispersive Raman spectra database of artists' materials (e-VISART database).

Raman spectroscopy has been widely applied in the analysis of different types of artwork. This technique is sensitive, reliable, non-destructive and can be used in situ. However, there are few references in the literature regarding specific Raman spectra libraries for the field of artwork analysis. In this paper, the development of two on-line databases with Fourier transform Raman (FT-Raman; 1064 nm) and dispersive Raman (785 nm) spectra of materials used in fine art is presented; both are implemented in the e-vibrational spectroscopic databases of artists' materials database (e-VISART). The database provides not only spectra, but also information about each pigment. It must be highlighted that for each pigment or material several spectra are available from different dealers. Some of the FT-Raman spectra available in the e-VISART database have not been published until now. Some examples in which the e-VISART database has been successfully used are presented.

Journal Article↗

Cryopreservation of adherent human embryonic stem cells.

Standard human embryonic stem (HES) cell cryopreservation methodologies, including slow freezing and vitrification of colonies in suspension, are plagued by poor viability and high differentiation rates upon recovery. To facilitate research studies and clinical applications of HES cells, we have developed a cryopreservation technique based on stabilizing HES colonies adherent to or embedded in a Matrigel matrix. This method increases cell viability by over an order of magnitude compared with cryopreservation in suspension and reduces differentiation. Loading adherent HES cells with the disaccharide trehalose prior to cryopreserving in a dimethylsulfoxide-containing cryoprotectant solution further improves cell viability under certain conditions. Our proposed approach has the potential to reduce the time required to amplify frozen stocks of HES cells, minimize risk of clonal selection during freeze-thaw cycles, and facilitate storage of HES cell clone libraries.

Biocompatible Materials↗

The construction of a cDNA expression library for the sheep scab mite Psoroptes ovis.

The need for alternative control strategies for sheep scab is critical. One approach is to develop vaccines based on 'concealed' antigens derived from Psoroptes ovis. This strategy requires the identification and characterisation of potential target antigens, which has been hampered by the problem of limited biological material for isolation of protein antigens. To aid the discovery of P. ovis antigens and to provide a resource for generating recombinant protein, we constructed a P. ovis cDNA expression library, using total RNA isolated from 250 mg of mixed-stage P. ovis and the Clontech SMART cDNA synthesis kit. The presence of P. ovis-specific sequences was confirmed using PCR amplification and sequencing of actin. The sequences of cDNA inserts from six random clones included one with high homology to the Dermatophagoides pteronyssinus (house dust mite) antigen p Dp15. This is a glutathione S-transferase known to be an important house dust mite antigen. We conclude that this library will be a useful tool for the identification of potential target antigens for the immunological control of P. ovis and to further our understanding of the pathology of sheep scab.

Amino Acid Sequence↗

TTY2: a multicopy Y-linked gene family.

Genes involved in human male sex determination and spermatogenesis are likely to be located on the Y chromosome. In an effort to identify Y-linked, testis-expressed genes, a cDNA selection library was generated by selecting testis cDNA with Y-cosmid clones. Resultant clones containing repetitive or vector material were eliminated, and 79 of the remaining clones were sequenced. Nineteen cDNAs showed homology with the TTY2 gene, and indicated that TTY2 is part of a large gene family. Screening of a panel of Y-linked cosmids revealed that the TTY2 gene family includes at least 26 members organized in 14 subfamilies. Further investigation revealed that TTY2 genes are arranged in tandemly arrayed clusters on both arms of the Y chromosome, and each gene comprises a series of tandemly arranged repeats. RT-PCR studies for two of these genes revealed that they are expressed in adult and fetal testis, as well as in the adult kidney. None of the genes investigated in detail contain an open reading frame. We conclude that the TTY2 gene family is composed of multiple copies, some of which may function as noncoding RNA transcripts and some may be pseudogenes.

Adult↗

SSH adequacy to preimplantation mammalian development: scarce specific transcripts cloning despite irregular normalisation.

BACKGROUND: SSH has emerged as a widely used technology to identify genes that are differentially regulated between two biological situations. Because it includes a normalisation step, it is used for preference to clone low abundance differentially expressed transcripts. It does not require previous sequence knowledge and may start from PCR amplified cDNAs. It is thus particularly well suited to biological situations where specific genes are expressed and tiny amounts of RNA are available. This is the case during early mammalian embryo development. In this field, few differentially expressed genes have been characterized from SSH libraries, but an overall assessment of the quality of SSH libraries is still required. Because we are interested in the more systematic establishment of SSH libraries from early embryos, we have developed a simple and reliable strategy based on reporter transcript follow-up to check SSH library quality and repeatability when starting with small amounts of RNA. RESULTS: Four independent subtracted libraries were constructed. They aimed to analyze key events in the preimplantation development of rabbit and bovine embryos. The performance of the SSH procedure was assessed through the large-scale screening of thousands of clones from each library for exogenous reporter transcripts mimicking either tester specific or tester/driver common transcripts. Our results show that abundant transcripts escape normalisation which is only efficient for rare and moderately abundant transcripts. Sequencing 1600 clones from one of the libraries confirmed and extended our results to endogenous transcripts and demonstrated that some very abundant transcripts common to tester and driver escaped subtraction. Nonetheless, the four libraries were greatly enriched in clones encoding for very rare (0.0005% of mRNAs) tester-specific transcripts. CONCLUSION: The close agreement between our hybridization and sequencing results shows that the addition and follow-up of exogenous reporter transcripts provides an easy and reliable means to check SSH performance. Despite some cases of irregular normalisation and subtraction failure, we have shown that SSH repeatedly enriches the libraries in very rare, tester-specific transcripts, and can thus be considered as a powerful tool to investigate situations where small amounts of biological material are available, such as during early mammalian development.

Animals↗