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PubMed2005

Preoperative education for orthopaedic patients: systematic review.

AIMS: This paper presents a systematic review whose aim was to describe the scope and methods of the current literature on preoperative patient education and to identify the effects of this education. BACKGROUND: Preoperative patient education is a common and important intervention in surgical nursing, yet there is very limited systematic evidence on its precise role. METHODS: The Medline, CINAHL, Eric, Psycinfo and Social Sciences Index databases and the Cochrane Library were searched, covering the period from the beginning of each database to April 2003. Studies were included if they concerned adult orthopaedic patients, preoperative nursing patient education and were based on randomized controlled or clinical trials. Meta-analysis was carried out where appropriate. RESULTS: We identified 11 articles involving 1044 participants. Most studies included one experimental and one control group; only two had more than one experimental and control group. The educational interventions varied widely, but the majority were based on written materials alone, or written materials in combination with other teaching methods. The most common outcome measures related to pain, knowledge, anxiety, exercises and length of stay, and the least common to self-efficacy and empowerment. The methodological quality of the studies varied. Almost all reported one or more statistically significant effects. Based on the findings of the meta-analysis, preoperative education appears to have some impacts on patients' anxiety and knowledge levels. CONCLUSIONS: The review clearly highlights the need for well-designed, methodologically sound research into the outcomes of patient education. It also points to the need to study patient education from the point of view of empowerment.

Anxiety↗
PubMed2005

Molecular analysis of human oral microbiota.

OBJECTIVES: The application of molecular, mainly 16S ribosomal RNA (rRNA)-based approaches enables researchers to bypass the cultivation step and has proven its usefulness in studying the microbial composition in a variety of ecosystems, including the human oral cavity. In this mini-review, we describe the impact of these culture-independent approaches on our knowledge of the ecology of the human oral cavity and provide directions for future studies that should emphasize the role of specific strains, species and groups of microbes in periodontal disease. MATERIALS AND METHODS: Recent findings are summarized to elucidate the relationship between periodontal disease and human oral microbiota, including as-yet-to-be-cultured organisms. RESULTS: The real-time polymerase chain reaction (PCR) method was developed to detect and quantify periodontopathic bacteria, such as Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis (formerly Bacteroides forsythus) and Treponema denticola. The checkerboard DNA-DNA hybridization technique allowed enumeration of large numbers of species in very large numbers of samples. 16S rRNA gene clone library analysis revealed the diversity of human oral microbiota and the existence of as-yet-to-be-cultured organisms that are presumed periodontal pathogens. In addition, terminal restriction fragment length polymorphism (T-RFLP) analysis was applied for assessment of diversity of human oral microbiota. CONCLUSION: Culture-independent approaches are useful for studying the microbial ecology in the human oral cavity and should be useful in the future to elucidate the etiology of periodontal disease.

DNA Probes↗
PubMed2003

Migration of volatile degradation products into ozonated water from plastic packaging materials.

Migration of volatile degradation products from poly(ethylene terephthalate) (PET) and high-density polyethylene (HDPE) bottles, polypropylene (PP) caps and ethyl vinyl acetate (EVA) liners into ozonated water was measured. Polymer strips were immersed in deionized and distilled water with ozone concentrations of 0.5, 2.5 and/or 5 mg kg(-1) inside 35-ml vials, which were clamp-sealed and stored at 40 degrees C for 10 days. A purge-and-trap unit was developed to extract volatile products from the ozonated water in vials. The extractables were trapped in an adsorbent tube and analysed using a GC-MS coupled with an automated thermal desorber (ATD). Mass spectra were interpreted by comparison with a NIST mass spectral library, and an internal standard method was used to quantify the extractables of interest. Several volatile compounds found in ozonated water that had been in contact with PP, EVA and HDPE polymers included butanal, pentanal, hexanal, heptanal, octanal, nonanal, 2,2-dimethyl propanal, 3-hexanone, 2-hexanone and heptanone. These compounds could cause off-taste and off-odour with a low organoleptic threshold. In general, the concentrations of these volatile compounds increased with an increased exposure to ozone. The highest concentration found was 14.1 +/- 0.6 microg kg(-1) for hexanal with a 5 mg kg(-1) ozone treatment of PP caps. Even at a treatment level of 5 mg kg(-1) ozone, which is greater than 10 times the current regulatory limits for bottled water, the extractables migrating from those polymers were within the levels permitted by the FDA. For the PET sample, no significant peaks were observed before or after ozonation. These results imply that PP caps containing EVA liners may be major sources of off-odour and taste in ozonated bottled water.

Disinfection↗
PubMed1993

Using a network menu and the UMLS Information Sources Map to facilitate access to online reference materials.

As computer technology advances, clinicians and biomedical researchers are becoming more dependent upon information from online databases and information systems. By using specially configured computer workstations and high-speed computer networks, it is now possible to access this information in a rapid and straightforward manner. To empower users by providing these capabilities, the authors are assembling a variety of network workstations to be located throughout Yale-New Haven Medical Center. At the heart of the workstation is NetMenu, a program designed to help users connect to a number of important online information systems, including a hospital order entry and results reporting system, a drug reference, bibliographic retrieval systems, and educational programs. In addition, as part of the National Library of Medicine's Unified Medical Language System (UMLS) project, the authors have developed a local prototype of the UMLS Information Sources Map (ISM) and a companion query assistant program to complement the NetMenu in helping users select and connect automatically to information services relevant to a particular question. The ISM query assistant draws from a listing of many online information sources accessible via local and international networks.

Computer Communication Networks↗
PubMed2002

Use of CD9 expression to enrich for porcine hematopoietic progenitors.

OBJECTIVE: The aim of this study was to develop novel markers for enrichment of hematopoietic progenitors from bone marrow of swine. MATERIALS AND METHODS: We previously showed that pig bone marrow contains a "side population" (SP) of Hoechst dye-effluxing cells that resembles the hematopoietic stem cell (HSC)-containing murine SP and therefore represents a putative pig stem cell population. We screened a panel of monoclonal antibodies for those that allowed positive or negative enrichment of porcine SP cells and tested one of these for enrichment of hematopoietic progenitors in short-term and long-term in vitro assays. We then screened an expression library to clone the gene whose product is recognized by this antibody. RESULTS: Among a panel of 35 monoclonal lines screened, we found three that were useful for positive enrichment of SP cells and seven for negative enrichment. The 4-6 monoclonal line, allowing around 10-fold negative enrichment of SP cells, recognized the product of the porcine CD9 gene. Hematopoietic progenitors measured by short-term colony-forming unit and long-term cobblestone area-forming cell assays were around 10-fold enriched in the CD9(negative/low) fraction and were significantly depleted in the CD9(high) fraction. CONCLUSIONS: The antibody against the porcine CD9 gene product may be of use for enrichment of porcine hematopoietic stem cells. This approach to identify novel markers for enrichment of hematopoietic progenitors may be applicable to other mammalian species.

Amino Acid Sequence↗
PubMed2003

A catalogue of Anopheles gambiae transcripts significantly more or less expressed following a blood meal.

The recent assembly of the Anopheles gambiae genome included the sequencing of ~80,000 EST from blood fed (BF) and non-blood fed (NBF) adult female mosquitoes to help find the exons and introns in the mosquito genome. These two EST libraries provided an insight into the differential gene expression resulting from the metabolically intense task of converting the massive blood meal into eggs. Previously, 168 genes have been identified to be either significantly more or significantly less transcribed after the blood meal [Science 298 (2002) 129]. Presently, 435 transcripts are described; these are grouped in supplemental tables by probable function to facilitate public access to these data and to help in designing postgenome experiments in the biology of Anopheles gambiae. The electronic tables and supplemental material are available at http://www.ncbi.nlm.nih.gov/projects/Mosquito/Ag-ESTs.

Animals↗
PubMed2004

AFLP targeting of the 1-cM region conferring the vrs1 gene for six-rowed spike in barley, Hordeum vulgare L.

Spike morphology is a key characteristic in the study of barley domestication, yield, and use. Multiple alleles at the vrs1 locus control the development and fertility of the lateral spikelets of barley. We developed five amplified fragment length polymorphism (AFLP) markers tightly linked to the vrs1 locus using well-characterized near-isogenic lines as plant materials. The AFLP markers were integrated into three different maps, in which 'Azumamugi' was used as the maternal parent. Of the three maps, Hordeum vulgare L. 'Azumamugi' x H. vulgare 'Golden Promise' showed recombination of the AFLP markers and the vrs1 locus (closest, 0.05 cM), providing the best mapping population for positional cloning of alleles at the vrs1 locus. Conversion of AFLP bands into polymorphic sequence-tagged sites (STSs) is necessary for further high-throughput genotype scoring and for bacterial artificial chromosome (BAC) library screening. We cloned and sequenced the five AFLP bands and synthesized primer pairs. PCR amplification generated DNAs of the same size from all four parental lines for each marker. Restriction endonuclease treatment of e40m36-1110/AccIII, e34m13-260/Psp1406I, e52m32-270/FokI, and e31m26-520/MnlI revealed fragment length polymorphisms between 'Azumamugi' and all the two-rowed parents. Allelism between the AFLPs and corresponding STS markers was confirmed genetically, indicating the usefulness of the STSs as genetic markers.

Alleles↗
PubMedDate not supplied

General inverse solid-phase synthesis method for C-terminally modified peptide mimetics.

Peptide mimetics are of considerable interest as bioactive agents and drugs. C-terminally modified peptide mimetics are of particular interest given the synthetic versatility of the carboxyl group and its derivatives. A general approach to C-terminally modified peptide mimetics, based on a urethane attachment strategy and amino acid t-butyl ester-based N-to-C peptide synthesis, is described. This approach is compatible with the reaction conditions generally employed for solution-phase peptide mimetic synthesis. To develop and demonstrate this approach, it was employed for the solid-phase synthesis of peptide trifluoromethyl ketones, peptide boronic acids, and peptide hydroxamic acids. The development of a versatile general approach to C-terminally modified peptides using readily available starting materials provides a basis for the combinatorial and parallel solid-phase synthesis of these peptide mimetic classes for bioactive agent screening and also provides a basis for the further development of solid-phase C-terminal functional group elaboration strategies.

Amino Acid Sequence↗
PubMed1990

Genome elimination in diploid and triploid Rana esculenta males: cytological evidence from DNA flow cytometry.

Cytological aspects of hemiclonal (meroclonal) inheritance in diploid and triploid males of the hybridogenetic frog Rana esculenta (Rana ridibunda x Rana lessonae) have been studied by DNA flow cytometry. The fact that the R. ridibunda genome contains 16% more DNA than the R. lessonae genome provides the ability to discern cells containing genomes of any species from the water-frog complex under study. Data are presented showing that elimination of the R. ridibunda genome occurs in hybridogenetic males from certain populations. In triploid males, the cytogenetic mechanism of hemiclonal inheritance is simpler than in diploids: after the elimination of a genome (always the genome in the minority in the triploid set; "homogenizing elimination"), no compensatory duplication of the remaining genetic material is necessary, as it is in diploids. The process of elimination can be visualized in triploid males by using DNA flow cytometry to identify cells in the special phase of the spermatogonial cell cycle that we termed the E phase.

Animals↗
PubMed1975

Cataloging audiovisual materials: a new dimension.

A new more comprehensive system for cataloging audiovisual materials is described. Existing audiovisual cataloging systems contain mostly descriptive information, publishers' or producers' summaries, and order information. This paper discusses the addition of measurable learning objectives to this standard information, thereby enabling the potential user to determine what can be learned from a particular audiovisual unit. The project included media in nursing only. A committee of faculty and students from the University of Alabama in Birmingham School of Nursing reviewed the materials. The system was field-tested at nursing schools throughout Alabama; the schools offered four different types of programs. The system and its sample product, the AVLOC catalog, were also evaluated by medical librarians, media specialists, and other nursing instructors throughout the United States.

Audiovisual Aids↗
PubMed1992

Nonradioactive in situ hybridisation of the translocation t(1;7) in myeloid malignancies.

Bone marrow cells of four patients with t(1;7) and myelodysplasia or acute myeloid leukemia were analyzed using nonradioactive in situ hydridisation. As probes, centromeric alphoid DNA sequences of chromosomes 1 and 7, a satellite DNA probe for 1q12, and chromosome-specific libraries of chromosomes 1 and 7 were used. The breakpoints of the t(1;7)(p11;p11) as determined by banding analysis could be studied more accurately, and the recently proposed designation t(1;7)(cen;cen) was confirmed in all four cases. Colocalization of alphoid DNA sequences of chromosomes 1 and 7 by double target in situ hybridisation was demonstrated in metaphase cells and also in interphase nuclei. The in situ hybridisation method described is applicable for the screening of peripheral blood cells or archival material.

Acute Disease↗
PubMed2003

Development and validation of a diagnostic microbial microarray for methanotrophs.

The potential of DNA microarray technology in high-throughput detection of bacteria and quantitative assessment of their community structures is widely acknowledged but has not been fully realised yet. A generally applicable set of techniques, based on readily available technologies and materials, was developed for the design, production and application of diagnostic microbial microarrays. A microarray targeting the particulate methane monooxygenase (pmoA) gene was developed for the detection and quantification of methanotrophs and functionally related bacteria. A microarray consisting of a set of 59 probes that covers the whole known diversity of these bacteria was validated with a representative set of extant strains and environmental clones. The potential of the pmoA microarray was tested with environmental samples. The results were in good agreement with those of clone library sequence analyses. The approach can currently detect less dominant bacteria down to 5% of the total community targeted. Initial tests assessing the quantification potential of this system with artificial PCR mixtures showed very good correlation with the expected results with standard deviations in the range of 0.4-17.2%. Quantification of environmental samples with this method requires the design of a reference mixture consisting of very close relatives of the strains within the sample and is currently limited by biases inherent in environmental DNA extraction and universal PCR amplification.

Bacteria↗
PubMed2007

Nanomechanical analysis of bone tissue engineering scaffolds.

Copolymers of (2-hydroxyethyl methacrylate) (HEMA) and methacrylamide monomers conjugated with amino acids were synthesized and crosslinked with ethylene glycol dimethacrylate. The resulting library of copolymers was mineralized in vitro using two distinct methods. In the first mineralization method, the copolymers were polymerized in the presence of a sub-micron hydroxyapatite (HA) suspension. In the second method, copolymers were mineralized with HA using a urea-mediated process. The mechanical properties of all of the copolymers, both mineralized and not, were determined using nanoindentation under both load and displacement control. A power law fit to the initial unloading curve was used to determine a reduced elastic modulus for each material. Between 30 and 300 indentations were performed on each material, and ANOVA analysis was run to determine the statistical significance of differences in modulus between samples. Using nanoindentation, the 22 different samples had reduced modulus values ranging from 840 MPa to 4.14 GPa. Aspartic acid-methacrylate (Asp-MA) copolymers were not distinguishable from the pHEMA control material. Polymerization in the presence of HA created a more uniform material than the urea method of mineralization. Several challenges and solutions encountered in the nanomechanical testing of soft, heterogeneous materials are discussed. These results demonstrate that with proper experimental design, the mechanical properties of tissue engineering scaffold materials based on polymer-ceramic composite materials can be determined using small samples and nanoindentation techniques.

Bone Substitutes↗
PubMed1994

Identification of a homeobox-containing gene with enhanced expression during soybean (Glycine max L.) somatic embryo development.

Homeotic gene are key 'switches' that control developmental processes. Homeotic genes containing the consensus 'homeobox' domain have been identified from a number of organisms including Drosophila melanogaster, Caenorhabditis elegans, Homo sapiens, and Zea mays. Although homeotic genes have been demonstrated to be important in embryo development of some insects, amphibians, and mammals, there are no reports of their involvement in plant embryogenesis. Here, we report the isolation and characterization of a cDNA clone for a homeobox-containing gene expressed in somatic embryos of soybean. The cDNA (Sbh1 for soybean homeobox-containing gene) was isolated using maize Knotted-1 (Kn1) cDNA as a heterologous probe. The Sbh1 cDNA clone is 1515 bp long which is the approximate size of its transcript. Within the homeodomain, the amino acid sequence of a helix-turn-helix structure, and invariant and conserved residues were identified. The deduced SBH1 protein shares a high amino acid identity with KN1 protein (47.0% overall and 87.5% for the homeodomain). Southern hybridization analysis indicated that Sbh1 is a member of a small gene family. The expression of Sbh1 is development- and tissue-specific. The transcript of Sbh1 was present in early-stage somatic embryos, increased prior to cotyledon formation and decreased thereafter. Sbh1 was weakly expressed in soybean stems and hypocotyl but was not detected in other plant tissues and nonembryogenic materials. The enhanced expression during embryogenesis, the homology with maize Kn1 gene, and the regulatory nature of homeodomain proteins suggest that the SBH1 protein plays an important role in plant embryo development.

Amino Acid Sequence↗
PubMed2005

Limitations of mRNA amplification from small-size cell samples.

BACKGROUND: Global mRNA amplification has become a widely used approach to obtain gene expression profiles from limited material. An important concern is the reliable reflection of the starting material in the results obtained. This is especially important with extremely low quantities of input RNA where stochastic effects due to template dilution may be present. This aspect remains under-documented in the literature, as quantitative measures of data reliability are most often lacking. To address this issue, we examined the sensitivity levels of each transcript in 3 different cell sample sizes. ANOVA analysis was used to estimate the overall effects of reduced input RNA in our experimental design. In order to estimate the validity of decreasing sample sizes, we examined the sensitivity levels of each transcript by applying a novel model-based method, TransCount. RESULTS: From expression data, TransCount provided estimates of absolute transcript concentrations in each examined sample. The results from TransCount were used to calculate the Pearson correlation coefficient between transcript concentrations for different sample sizes. The correlations were clearly transcript copy number dependent. A critical level was observed where stochastic fluctuations became significant. The analysis allowed us to pinpoint the gene specific number of transcript templates that defined the limit of reliability with respect to number of cells from that particular source. In the sample amplifying from 1000 cells, transcripts expressed with at least 121 transcripts/cell were statistically reliable and for 250 cells, the limit was 1806 transcripts/cell. Above these thresholds, correlation between our data sets was at acceptable values for reliable interpretation. CONCLUSION: These results imply that the reliability of any amplification experiment must be validated empirically to justify that any gene exists in sufficient quantity in the input material. This finding has important implications for any experiment where only extremely small samples such as single cell analyses or laser captured microdissected cells are available.

Analysis of Variance↗
PubMed2005

Selection of gonadotrophin surge attenuating factor phage antibodies by bioassay.

BACKGROUND: We aimed to combine the generation of "artificial" antibodies with a rat pituitary bioassay as a new strategy to overcome 20 years of difficulties in the purification of gonadotrophin surge-attenuating factor (GnSAF). METHODS: A synthetic single-chain antibody (Tomlinson J) phage display library was bio-panned with partially purified GnSAF produced by cultured human granulosa/luteal cells. The initial screening with a simple binding immunoassay resulted in 8 clones that were further screened using our in-vitro rat monolayer bioassay for GnSAF. Initially the antibodies were screened as pooled phage forms and subsequently as individual, soluble, single-chain antibody (scAbs) forms. Then, in order to improve the stability of the scAbs for immunopurification purposes, and to widen the range of labelled secondary antibodies available, these were engineered into full-length human immunoglobulins. The immunoglobulin with the highest affinity for GnSAF and a previously described rat anti-GnSAF polyclonal antiserum was then used to immunopurify bioactive GnSAF protein. The two purified preparations were electrophoresed on 1-D gels and on 7 cm 2-D gels (pH 4-7). The candidate GnSAF protein bands and spots were then excised for peptide mass mapping. RESULTS: Three of the scAbs recognised GnSAF bioactivity and subsequently one clone of the purified scAb-derived immunoglobulin demonstrated high affinity for GnSAF bioactivity, also binding the molecule in such as way as to block its bioactivity. When used for repeated immunopurification cycles and then Western blot, this antibody enabled the isolation of a GnSAF-bioactive protein band at around 66 kDa. Similar results were achieved using the rat anti-GnSAF polyclonal antiserum. The main candidate molecules identified from the immunopurified material by excision of 2-D gel protein spots was human serum albumin precursor and variants. CONCLUSION: This study demonstrates that the combination of bioassay and phage display technologies is a powerful tool in the study of uncharacterised proteins that defy conventional approaches. In addition, we conclude that these data support suggestions that GnSAF may be structurally related to serum albumin or very tightly bound to serum albumin.

Animals↗
PubMed1985

Expression of the L11 neuropeptide gene in the Aplysia central nervous system.

Neuron L11 in the abdominal ganglion of Aplysia californica is thought to be both cholinergic and peptidergic. In previous studies, we isolated a cDNA clone encoding the precursor for an L11 secreted protein(s) by differentially screening an abdominal ganglion cDNA library. We now report the isolation of genomic clones encoding the L11 cDNA sequences. Analysis of these clones reveals that the gene is present in a single copy per haploid genome. RNA blotting and cDNA cloning demonstrate that the L11 gene is expressed not only in the abdominal ganglion but in the head ganglia as well. To define the positions of cells expressing this gene and to follow their processes, we raised antibodies to synthetic peptides defined by the cDNA sequence. Histochemistry revealed about 100 neurons containing immunoreactive material. These cells arborize in the neuropil and are distributed throughout the central nervous system, representing about 0.5% of the Aplysia central neurons. In addition, cells in the abdominal ganglion send processes to the mantle floor at the base of the gill via the genital and branchial nerves. Our data suggest that this network of cells expresses the single L11 peptide gene.

Animals↗
PubMed1977

Characterization of the major diazo-positive pigments in bile of homozygous Gunn rats.

Bilinoid pigments in bile of homozygous Gunn rats (jj) were analysed either after formation of dipyrrolic ethyl anthranilate azo derivatives or as the unmodified parent tetrapyrroles. 1. T.l.c. of the azo derivatives revealed seven major unconjugated components which were structurally characterized by chemical tests, spectrophotometry and mass spectrometry. In addition, two minor components were identified as azodipyrrole (A+B)-glucoside and azodipyrrole (A+B)-beta-d-glucuronide. 2. Extraction and t.l.c. of the tetrapyrrolic pigments showed 13 major yellow diazo-positive bands. Four of them, accounting for 59% of total diazo-positive material, were identified as unconjugated bilirubin-IXalpha, -IXbeta, -IXgamma and -IXdelta. A fifth band (16%) was characterized as a mixture of two isomeric monohydroxyl derivatives and another band (8%) as a dihydroxyl derivative of bilirubin-IXalpha. 3. Although unconjugated bilirubin-IXalpha constitutes one-third of total diazo-positive material in bile of our strain of Gunn rats, the daily amount excreted represented only about 3-4% of daily bilirubin production. 4. Phototherapy caused a 2.2-fold increase in the biliary output of diazo-positive bilinoids, but did not affect markedly their composition. However, an additional diazo-negative pigment, accounting for one-third of total yellow colour, was observed but was not identified. Mass-spectral data on two dipyrrolic azopigments have been deposited as Supplementary Publication SUP 50076 (3 pages) with the British Library Lending Division, Boston Spa, Wetherby, W. Yorkshire LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1977) 161, 1.

Animals↗