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Cloning of human, murine, and marsupial keratin 7 and a survey of K7 expression in the mouse.

Keratins are cytoplasmic intermediate filament proteins expressed by epithelial cells. Keratin 7 (K7) is expressed in a wide range of epithelial structures in humans. We have cloned and fully sequenced the human and mouse K7 genes and mRNAs, and the K7 mRNA from the marsupial Potorous tridactylis, from which the widely used simple epithelial cell lines PtK1 and PtK2 are derived. Percentage identity plots comparing the mouse and human genomic sequences revealed a number of conserved CpG islands associated with the K7 gene. There was considerable conservation of introns between the two species, which may indicate the presence of intronic regulatory elements. Only the most proximal 500bp of the promoter was conserved, although an additional conserved sequence island was found 2-3kb upstream. Protein sequence comparisons between the three species allowed identification of conserved regions of the keratin variable domains that may be candidates for protein-protein interactions and/or regulatory modification. From the mouse sequence, we generated a polyclonal rabbit antibody specific for murine K7. This antibody was used to perform a survey of K7 expression in the mouse. The expression pattern was similar to the reported human distribution, with substantial expression observed in lung, bladder, mesothelium, hair follicle, and ductal structures. We also noted previously unreported expression of K7 in the gastrointestinal tract and filiform papillae of the tongue and specific K7 expression in a range of "hard" epithelial tissues. The distribution of K7 in mouse and availability of genomic sequence from the 129/Sv mouse strain will allow the generation and analysis of transgenic mice expressing mutant forms of K7 and to predict the phenotype of human genetic disorders caused by mutations in this keratin.

Amino Acid Sequence↗

Preventing postpartum depression part I: a review of biological interventions.

OBJECTIVE: This paper critically reviews the literature to determine the current state of scientific knowledge concerning the prevention of postpartum depression (PPD) from a biological perspective. METHODS: The criteria used to evaluate the interventions were derived from the standardized methodology developed by the Canadian Task Force on Preventive Health Care. Databases searched for this review include Medline, PubMed, Cinahl, PsycINFO, Embase, ProQuest, the Cochrane Library, and the World Health Organization Reproductive Health Library. Studies selected were peer-reviewed English-language articles published between January 1, 1966, and December 31, 2003. RESULTS: Seven studies that met criteria were examined. These studies focused on evaluating the preventive effect of antidepressant medication, estrogen and progesterone therapy, thyroid therapy, docosahexanoic acid, and calcium supplementation. Although some of these interventions have been examined rigorously for depression unrelated to childbirth, methodological study limitations render intervention efficacy equivocal for PPD; thus, there is limited strong evidence to guide practice or policy recommendations. CONCLUSIONS: Despite the recent upsurge of interest in this area, many questions remain unanswered, which has several implications for research.

Adult↗

Accessing the biomedical literature: the South Dakota Med-Fax Network.

Librarians developed a plan for a medical information network based upon the needs of South Dakota physicians as revealed in a 1984 survey. The South Dakota Med-Fax Network received private funding in 1987 and became operational in 1988. The Network incorporates interactive simultaneous remote on-line database searching (ISRS) and telefacsimile transmission to expedite the identification and delivery of medical literature to South Dakota physicians. At present, the Network is composed of sixteen institutional sites. Individual physicians may establish their own sites if they provide an IBM compatible personal computer with a modem and purchase a telecommunications software package. The Network will provide training and follow-up support. Use patterns to date demonstrate that telefacsimile transmission is instrumental in reducing significantly the time necessary to deliver journal literature and that ISRS searching is the method of choice in the Network's remote sites.

Humans↗

Analysis of surface properties of human cancer cells using derivatized beads.

Standard histochemical analysis of cells and tissues generally involves procedures that utilize a relatively small number of probes such as dyes, and generally requires hours or days to process. Our laboratory has developed a novel method for histochemical surveys of cell surface properties that utilizes a large number of probes (derivatized agarose beads) and takes seconds or minutes to accomplish. In this study, 4 human cell lines (CCL-255 (LS123) human colon cancer cells that are non-tumorigenic in nude mice; CRL-1459 (CCD-18CO) human colon endothelial cells that are non-malignant; CCL-220 (COLO 320DM) human colon cancer cells that are tumorigenic in nude mice; and HTB-171 (NCI H446) human lung carcinoma cells) were tested for their ability to bind to agarose beads derivatized with 51 different molecules. There were statistically significant differences in binding of the 4 cell types to all of the 51 types of beads, but 15 types of beads showed dramatic differences in binding to one or more of the 4 cell types. For example, only HTB-171 (NCI H446) bound to p-aminophenyl-beta-D-glucopyranoside-derivatized beads and only CCL-220 (COLO 320DM) bound to L-tyrosine-derivatized beads. The specificity of cell-bead binding was examined by performing assays in the presence or absence of exogenously added compounds in hapten-type of inhibition experiments. This assay, that utilizes large numbers of novel probes, may help in the development of new libraries of surface properties of specific cell types, with differing degrees of malignancy, that at this time could not be developed by using other available technologies.

Animals↗

Effect of an evidence-based answering service on GPs and their patients: a pilot study.

OBJECTIVES: For general practitioners (GPs), an important obstacle to practising evidence-based medicine is lack of time. An evidence-based answering service was developed that took over searching and appraisal of medical evidence from the GPs. GPs sent in questions, and the informationist formulated the answers. Our objectives were to find out if such an evidence-based answering service was feasible, including assessing the effect of the answers on GPs and their patients, as reported by the GPs. METHODS: After attending a workshop on building well-formulated questions from daily practice, the GPs sent in questions to the informationist. The literature was searched, the relevant information was appraised, and the answers to the questions were formulated. With a questionnaire, the effect of the answers on the GP and the patient was assessed, as well as the perceived barriers to implementing the answers. RESULTS: From 26 GPs, 61 questions were received. For 12% of questions, information was found with the highest evidence level, while for 36%, no information was found. However, for 89% of the questions for which no information could be found, the answer 'no information found' did have an effect on the GP concerned. In total, 81% of all the answers had an effect on the GP, and, according to the GP, 52% had an effect on the patient. Few barriers to implementing the evidence were perceived. Most of the answers were found in Pubmed/Medline, the Cochrane Library and Embase. CONCLUSIONS: This study indicates that an evidence-based answering service can have an impact on GPs and their patients. Librarians can provide an evidence-based answering service for GPs and their patients. The evidence-based answering service for GPs in this study had an impact on 81% of the GPs and on 52% of their patients. Although for one-third of the questions no evidence-based answer was found, this message in itself had an impact on 89% of the GPs. An informationist as mediator between medical information and doctors can save doctors' time.

Evidence-Based Medicine↗

Human gastric adenocarcinoma cathepsin B: isolation and sequencing of full-length cDNAs and polymorphisms of the gene.

Four full-length cDNA clones coding for preprocathepsin B were isolated from a human gastric adenocarcinoma cDNA library (AGS 1-6-30-1) and analyzed for possible sequence modifications that might be linked to altered intracellular trafficking and secretion of cathepsin B (CTSB) in malignant tumors. Comparison of AGS 1-6-30-1 cDNAs with human kidney/hepatoma cDNAs revealed: (1) three potential N-glycosylation sites instead of two, (2) a nucleotide (nt) substitution in the coding region for the propeptide from GTG to CTG which would result in a Val26-->Leu change, (3) three silent nt replacements in the coding region for the mature protein, (4) five single-nt differences in the 5'- and 3'-UTR (untranslated regions), (5) heterogeneity in the 5'-UTR, and (6) a 10-bp insertion in the 3'-UTR. The 10-bp insertion in the 3'-UTR may alter the stability of CTSB mRNA transcripts and thereby the expression of CTSB. These clones should be useful for expressing human tumor CTSB and analyzing the function of this enzyme in malignant progression. Two restriction-fragment length polymorphisms (RFLPs), EcoRI and TaqI, were detected by Southern blot analysis of genomic DNA from 36 unrelated Caucasians. Inheritance and distribution of the EcoRI alleles (13.0 and 11.0 kb) and the TaqI alleles (5.7 and 4.6 kb) indicated they were independent polymorphisms. In contrast to the EcoRI alleles of 13.0 and 11.0 kb observed in the population survey, genomic DNA from two AGS gastric adenocarcinoma subclones revealed two EcoRI alleles of 13.0 and 7.8 kb.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenocarcinoma↗

OQAFMA Querying agent for the Foundational Model of Anatomy: a prototype for providing flexible and efficient access to large semantic networks.

The development of large semantic networks, such as the UMLS, which are intended to support a variety of applications, requires a flexible and efficient query interface for the extraction of information. Using one of the source vocabularies of UMLS as a test bed, we have developed such a prototype query interface. We first identify common classes of queries needed by applications that access these semantic networks. Next, we survey StruQL, an existing query language that we adopted, which supports all of these classes of queries. We then describe the OQAFMA Querying Agent for the Foundational Model of Anatomy (OQAFMA), which provides an efficient implementation of a subset of StruQL by pre-computing a variety of indices. We describe how OQAFMA leverages database optimization by converting StruQL queries to SQL. We evaluate the flexibility and efficiency of our implementation using English queries written by anatomists. This evaluation verifies that OQAFMA provides flexible, efficient access to one such large semantic network, the Foundational Model of Anatomy, and suggests that OQAFMA could be an efficient query interface to other large biomedical knowledge bases, such as the Unified Medical Language System.

Abstracting and Indexing↗

Towards a consensus on datasets and evaluation metrics for developing B-cell epitope prediction tools.

A B-cell epitope is the three-dimensional structure within an antigen that can be bound to the variable region of an antibody. The prediction of B-cell epitopes is highly desirable for various immunological applications, but has presented a set of unique challenges to the bioinformatics and immunology communities. Improving the accuracy of B-cell epitope prediction methods depends on a community consensus on the data and metrics utilized to develop and evaluate such tools. A workshop, sponsored by the National Institute of Allergy and Infectious Disease (NIAID), was recently held in Washington, DC to discuss the current state of the B-cell epitope prediction field. Many of the currently available tools were surveyed and a set of recommendations was devised to facilitate improvements in the currently existing tools and to expedite future tool development. An underlying theme of the recommendations put forth by the panel is increased collaboration among research groups. By developing common datasets, standardized data formats, and the means with which to consolidate information, we hope to greatly enhance the development of B-cell epitope prediction tools.

Animals↗

Building a knowledge base to support a digital library.

As part of an effort to develop a knowledge base to support searching online medical literature according to individual needs, we have studied the possibility of using the co-occurrence of MeSH terms in MEDLINE citations associated with the search strategies optimal for evidence based medicine to automated construction of a knowledge base. This study evaluates the relevance of the relationships between the semantic relationship pairs generated by the process, and the clinical validity of the semantic types involved in the process. From the semantic pairs proposed by our method, a group of clinicians judge sixty percent to be relevant. The remaining forty percent included semantic types considered unimportant by clinicians. The knowledge extraction method showed reasonable results. We believe it can be appropriate for the task of retrieving information from the medical record in order to guide users during a searching and retrieval process. Future directions include the validation of the knowledge, based on an evaluation of system performance.

Algorithms↗

The behavior of preschoolers during and after brief surgical hospitalizations.

This qualitative study describes the behavior of preschool children during and after very brief hospitalizations for tonsillectomies and/or adenoidectomies. Structured, open-ended home interviews were conducted with 23 parents 5 to 7 days after their children's hospital discharge. Parents were asked to describe their children's behaviors during hospitalization and the first week at home. Approximately half of the parents and children had attended an optional surgical preadmission teaching program. The remaining parents had prepared their children for hospitalization using toy doctor kits and library books. Preoperatively, children who had attended the program behaved similarly to those who had not: They either cooperated fully, resisted initially, or completely refused to cooperate with admission procedures. Most children separated easily from their parents to enter the operating room, whereas some (n = 5) demonstrated great anxiety. However, immediately after surgery, most children who had not attended the program appeared more distressed than those who had attended. Finally, despite their relatively minor surgeries and brief hospitalizations, after discharge the majority of children (83%), regardless of whether they had attended the program, exhibited signs of distress. In comparison with their prehospitalization behavior, parents reported that their children were more irritable, had more nightmares, had appetite changes, and experienced more separation anxiety. Implications for nursing practice and research are discussed.

Adenoidectomy↗

Sequence diversity and large-scale typing of SNPs in the human apolipoprotein E gene.

A common strategy for genotyping large samples begins with the characterization of human single nucleotide polymorphisms (SNPs) by sequencing candidate regions in a small sample for SNP discovery. This is usually followed by typing in a large sample those sites observed to vary in a smaller sample. We present results from a systematic investigation of variation at the human apolipoprotein E locus (APOE), as well as the evaluation of the two-tiered sampling strategy based on these data. We sequenced 5.5 kb spanning the entire APOE genomic region in a core sample of 72 individuals, including 24 each of African-Americans from Jackson, Mississippi; European-Americans from Rochester, Minnesota; and Europeans from North Karelia, Finland. This sequence survey detected 21 SNPs and 1 multiallelic indel, 14 of which had not been previously reported. Alleles varied in relative frequency among the populations, and 10 sites were polymorphic in only a single population sample. Oligonucleotide ligation assays (OLA) were developed for 20 of these sites (omitting the indel and a closely-linked SNP). These were then scored in 2179 individuals sampled from the same three populations (n = 843, 884, and 452, respectively). Relative allele frequencies were generally consistent with estimates from the core sample, although variation was found in some populations in the larger sample at SNPs that were monomorphic in the corresponding smaller core sample. Site variation in the larger samples showed no systematic deviation from Hardy-Weinberg expectation. The large OLA sample clearly showed that variation in many, but not all, of OLA-typed SNPs is significantly correlated with the classical protein-coding variants, implying that there may be important substructure within the classical epsilon 2, epsilon 3, and epsilon 4 alleles. Comparison of the levels and patterns of polymorphism in the core samples with those estimated for the OLA-typed samples shows how nucleotide diversity is underestimated when only a subset of sites are typed and underscores the importance of adequate population sampling at the polymorphism discovery stage. [The sequence data described in this paper have been submitted to the GenBank data library under accession no. AF261279.]

Alleles↗

Factors associated with successful answering of clinical questions using an information retrieval system.

OBJECTIVES: Despite the growing use of online databases by clinicians, there has been very little research documenting how effectively they are used. This study assessed the ability of medical and nurse-practitioner students to answer clinical questions using an information retrieval system. It also attempted to identify the demographic, experience, cognitive, personality, search mechanics, and user-satisfaction factors associated with successful use of a retrieval system. METHODS: Twenty-nine students completed questionnaires of clinical and computer experience as well as tests of cognitive abilities and personality type. They were then administered three clinical questions to answer in a medical library setting using the MEDLINE database and electronic and print full-text resources. RESULTS: Medical students were able to answer more questions correctly than nurse-practitioner students before and after searching, but both had comparable improvements in the number of correct questions before and after searching. Successful ability to answer questions was also associated with having experience in literature searching and higher standardized test-score percentiles. CONCLUSIONS: Medical and nurse-practitioner students obtained comparable benefits in the ability to answer clinical questions from use of the information retrieval system. Future research must examine strategies that improve successful search and retrieval of clinical questions posed by clinicians in practice.

Adult↗

Living conditions of adults with intellectual disabilities from a gender perspective.

BACKGROUND: The role of gender has been a neglected issue in research on intellectual disability (ID). People with ID are generally treated as a homogenous group that are largely categorized by their level of ID. This study compared living conditions of women and men with ID and related the results to similarities and differences among the general population in corresponding age groups. METHODS: Persons with ID born in Uppsala County between 1959 and 1974 constituted the study sample. Information on the living conditions of 110 persons with ID was collected using questionnaires completed by relatives and staff. Information on living conditions of the general population was obtained through national welfare statistics conducted by Statistics Sweden (SCB). RESULTS: In both samples corresponding diversities were revealed for type of employment/daily activities, where women worked in traditional female job sectors and men were occupied with traditional male jobs. Women and men with ID participated to about the same extent in recreational and cultural activities and on only four of the 19 activities listed in the questionnaire (visits to the cinema and library, reading books and practising hobbies alone) significant differences were observed. Among women and men in the general population, we found gender-related differences in 13 of the activities listed. However, with the exception of women more frequently visiting the library and reading books, the two samples demonstrated no corresponding gender-related differences. For the remaining six domains (finances, family and social relations, housing, transport, community participation and personal safety), no differences were noted between women and men with ID. This finding contrasted sharply with the differences found between women and men in the general population. CONCLUSIONS: Surprisingly, the comparison yielded few differences in living conditions between women and men with ID compared with those found in women and men of the general population. This finding suggests that people with ID were treated as gender-neutral persons rather than as women and men with individual preferences and needs. Thus, it appears that having ID is a more important determinant than gender regarding living conditions for women and men with ID.

Activities of Daily Living↗

Multiple copies of nodD in Rhizobium tropici CIAT899 and BR816.

Rhizobium tropici strains are able to nodulate a wide range of host plants: Phaseolus vulgaris, Leucaena spp., and Macroptilium atropurpureum. We studied the nodD regulatory gene for nodulation of two R. tropici strains: CIAT899, the reference R. tropici type IIb strain, and BR816, a heat-tolerant strain isolated from Leucaena leucocephala. A survey revealed several nodD-hybridizing DNA regions in both strains: five distinct regions in CIAT899 and four distinct regions in BR816. Induction experiments of a nodABC-uidA fusion in combination with different nodD-hybridizing fragments in the presence of root exudates of the different hosts indicate that one particular nodD copy contributes to nodulation gene induction far more than any other nodD copy present. The nucleotide sequences of both nodD genes are reported here and show significant homology to those of the nodD genes of other rhizobia and a Bradyrhizobium strain. A dendrogram based on the protein sequences of 15 different NodD proteins shows that the R. tropici NodD proteins are linked most closely to each other and then to the NodD of Rhizobium phaseoli 8002.

Amino Acid Sequence↗

Physician use of the curbside consultation to address information needs: report on a collective case study.

PURPOSE: The author reports key findings from a doctoral dissertation investigating what the curbside consultation is, how and why physicians use it, and what the implications for health sciences library services might be. SETTINGS/INFORMANTS: Primary informants included sixteen primary care physicians at six sites in one Midwestern state. Additional informants included twenty-eight specialists and subspecialists identified by the primary informants as colleagues who provided curbside consultations. METHODS: Qualitative research methods were used, including field observations, formal and informal interviews, and conversations with peer review physicians. RESULTS: Despite a lack of consensus about what constitutes a "good" curbside consultation, physician informants reported that curbside consultations were part of their medical education and that they continued to take part in them for a number of reasons. Tacit rules govern curbside consultation interactions, and negative consequences result when the rules are misunderstood or not observed. DISCUSSION/CONCLUSION: Acknowledging and understanding physicians' use of the curbside consultation to obtain and construct knowledge may suggest new ways for health sciences librarians to work with physicians in locating, diffusing, and disseminating clinical information.

Education, Medical, Continuing↗

Assessment of MLA assertiveness training for librarians: students' behavior changes after taking C.E. 669, Assertiveness and Human Relations Skills.

C.E. 669, offered for three years, was MLA's first personal development continuing education course. Participants were asked to respond to a follow-up survey about their current assertive philosophy, awareness, and behavioral repertoire. They were also assessed by the Librarian's Assertiveness Inventory, a tool that indicates patterns of behavioral responses, for which some normative data were available from librarians who had not taken the class. Class participants were found to be more likely to behave assertively and less likely to behave aggressively than librarians who had not taken the class. In nine out of ten situational categories, librarians who had taken the class demonstrated assertive response patterns. They also reported remembering and using more than one third of the behavioral techniques covered in class, with particular use of the nonverbal techniques. A majority saw themselves as more assertive since taking the class, although most believed that others viewed them much the same as they had prior to the class. Positive progress toward reaching specific behavioral goals was also reported.

Assertiveness↗

Community hospitals--the place of local service provision in a modernising NHS: an integrative thematic literature review.

BACKGROUND: Recent developments within the United Kingdom's (UK) health care system have re-awakened interest in community hospitals (CHs) and their role in the provision of health care. This integrative literature review sought to identify and assess the current evidence base for CHs. METHODS: A range of electronic reference databases were searched from January 1984 to either December 2004 or February 2005: Medline, Embase, Web of Knowledge, BNI, CINAHL, HMIC, ASSIA, PsychInfo, SIGLE, Dissertation Abstracts, Cochrane Library, Kings Fund website, using both keywords and text words. Thematic analysis identified recurrent themes across the literature; narrative analyses were written for each theme, identifying unifying concepts and discrepant issues. RESULTS: The search strategy identified over 16,000 international references. We included papers of any study design focussing on hospitals in which care was led principally by general practitioners or nurses. Papers from developing countries were excluded. A review of titles revealed 641 potentially relevant references; abstract appraisal identified 161 references for review. During data extraction, a further 48 papers were excluded, leaving 113 papers in the final review. The most common methodological approaches were cross-sectional/descriptive studies, commentaries and expert opinion. There were few experimental studies, systematic reviews, economic studies or studies that reported on longer-term outcomes. The key themes identified were origin and location of CHs; their place in the continuum of care; services provided; effectiveness, efficiency and equity of CHs; and views of patients and staff. In general, there was a lack of robust evidence for the role of CHs, which is partly due to the ad hoc nature of their development and lack of clear strategic vision for their future. Evidence for the effectiveness and efficiency of the services provided was limited. Most people admitted to CHs appeared to be older, suggesting that admittance to CHs was age-related rather than condition-related. CONCLUSION: Overall the literature surveyed was long on opinion and short of robust studies on CHs. While lack of evidence on CHs does not imply lack of effect, there is an urgent need to develop a research agenda that addresses the key issues of health care delivery in the CH setting.

Health Services Research↗

Gene profiling techniques and their application in angiogenesis and vascular development.

The analysis of gene expression in specific tissues and physiological processes has evolved over the last 20 years from the painstaking identification of selected genes to the relatively efficient and open-ended surveying of potentially all genes expressed in a tissue. Current art for gene discovery includes the use of large-scale arrays of cDNA sequences or oligonucleotides, and molecular 'tagging' techniques such as GeneCalling and SAGE. Common to each of these techniques is a reliance on the increasingly comprehensive databases of human and mouse EST and full-length gene sequences. Early efforts to characterize candidate genes were limited by their narrow scope, while current efforts are confounded by the enormous volume of data returned. Sophisticated software tools are an integral part of the analysis, helping to organize information into coherent groups with temporal or functional similarity. These techniques, in conjunction with the continued analysis of human genetic syndromes, transgenic, and knockout mice, have driven genetic analysis of angiogenesis and vascular development from describing which individual genes are involved to defining the outlines of regulatory networks.

Animals↗