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PubMed2004

Rapid analysis of protein-nucleic acid complexes using MALDI TOF mass spectrometry and ion pair reverse phase liquid chromatography.

The RuvABC resolvasome of Escherichia coli typifies nucleoprotein complexes involved in genetic transactions. This molecular assembly catalyses the resolution of Holliday junctions that arise during genetic recombination and DNA repair. This process involves two key steps: branch migration, catalysed by the RuvB protein that is targeted to the Holliday junction by the structure specific RuvA protein, and resolution, which is catalysed by the RuvC endonuclease. We have used matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS) to rapidly identify the binding of RuvA to an immobilised synthetic Holliday junction; unambiguous identification was verified using tryptic digest of the bound protein. In conjunction with a novel fluorescent-based technique incorporating ion pair reverse phase liquid chromatography, a "footprint" of the RuvA:Holliday complex was obtained. These two complementary techniques offer a generic approach to the analysis of nucleoprotein complexes.

Base Sequence↗
PubMed2001

Managed competition, governmentality and institutional response in the United Kingdom.

This article traces the use of managed competition policy to transform the NHS from an administered public service to a set of interlocking markets and contracts. It reviews the overlooked origins of managed competition in the new managerialism and explains the relationship between managed competition and the cost crisis of the NHS by extending Foucault's concept of governmentality to revise the concept of the state. The paper then describes how the government structured health care markets, using managed competition as an instrument of governmentality. It summarises institutional responses by health authorities, hospital trusts, and GP fundholders. The terms "master institution", "dictated competition" and "coercive partnering" are introduced as new concepts for economic sociology and as strategies of governmentality. Implementation, however, led to resistance, opposition and eventual abandonment of managed competition as too disruptive and costly. Yet, this analysis contends, managed competition has left an enduring legacy of accountability to purchasers in economic terms such as efficiency, transaction costs, and cost effectiveness. The policies of the new government are based on coercive partnering and doctor-based "commissioning". This and the Internet imply revolutionary changes for the health professions and the delivery of health care services through networks of moebius-strip organisations interacting in flexible sequences and subject to communitarian pressures.

Contract Services↗
PubMed2003

Human proliferating cell nuclear antigen, poly(ADP-ribose) polymerase-1, and p21waf1/cip1. A dynamic exchange of partners.

We addressed the analysis of the physical and functional association of proliferating cell nuclear antigen (PCNA), a protein involved in many DNA transactions, with poly(ADP-ribose) polymerase (PARP-1), an enzyme that plays a crucial role in DNA repair and interacts with many DNA replication/repair factors. We demonstrated that PARP-1 and PCNA co-immunoprecipitated both from the soluble and the DNA-bound fraction isolated from S-phase-synchronized HeLa cells. Immunoprecipitation experiments with purified proteins further confirmed a physical association between PARP-1 and PCNA. To investigate the effect of this association on PARP-1 activity, an assay based on the incorporation of radioactive NAD was performed. Conversely, the effect of PARP-1 on PCNA-dependent DNA synthesis was assessed by a DNA polymerase delta assay. A marked inhibition of both reactions was found. Unexpectedly, PARP-1 activity also decreased in the presence of p21waf1/cip1. By pull-down experiments, we provided the first evidence for an association between PARP-1 and p21, which involves the C-terminal part of p21 protein. This association was further demonstrated to occur also in vivo in MNNG (N-methyl-N'-nitro-N-nitrosoguanidine)-treated human fibroblasts. These observations suggest that PARP-1 and p21 could cooperate in regulating the functions of PCNA during DNA replication/repair.

Cells, Cultured↗
PubMed1996

Coping styles among adolescent competitive athletes.

The use of approach and avoidance coping styles and task-focused and emotion-focused coping strategies in competitive sport was explored. Four hundred twenty-one adolescent males from New South Wales, Australia, who were currently competing in team sports indicated their usual responses to each of 8 acute stressors commonly experienced in sport, using a 128-item inventory. The reliability coefficient (Cronbach's alpha) for each stressor ranged from .81 to .92. Twenty-six of the original 128 items on the inventory were retained, on the basis of factor analysis. Correlations between stressors indicated that coping styles were a function of type of stressor, providing support for the transactional model. Goodness of fit was high (.87). The present results partially support the construct of coping style among adolescent-aged sports competitors.

Adaptation, Psychological↗
PubMed1993

Mouse gelatinase B. cDNA cloning, regulation of expression and glycosylation in WEHI-3 macrophages and gene organisation.

Gelatinase B is a regulated matrix metalloproteinase with an important role in the remodelling of extracellular matrices and of basement membranes. To study the structure and function of gelatinase B in the mouse, the cDNA was cloned from a macrophage cell line (WEHI-3). Using this cDNA, a cosmid clone with the mouse gene was isolated. The complete gene (8 kbp) was sequenced and compared with the human gene structure. There was 78% similarity at the cDNA level and the exon/intron structure of the murine gene was similar to the human counterpart. At the 5' untranslated side, 1200 bp of the promoter/enhancer region were sequenced and found to contain several transacting-factor-binding sites. The mRNA transcription-initiation site was determined by non-isotopic primer-extension analysis. Polymerase-chain-reaction amplification of cDNAs yielded indirect evidence for a reverse-transcription stop in WEHI-3 cell mRNA. The DNA-derived mouse-protein structure exhibited 82% similarity with the human one. This similarity was functionally reflected by cross-reactivity of the mouse protein with an antiserum against human gelatinase B. The production of murine gelatinase B was studied at the protein level by zymography and at the mRNA level by Northern blot analysis. In WEHI-3 cells the gelatinase B protein is induced by bacterial lipopolysaccharide, phorbol ester, double-stranded RNA and the cytokine interleukin-1. Regulation of activity and structural heterogeneity of gelatinase B in WEHI-3 cells were shown to occur at the gene regulatory level, by expression of the matrix metalloproteinase inhibitor TIMP-1, and by glycosylation of the secreted protein.

Amino Acid Sequence↗
PubMed2004

Eukaryotic MCM proteins: beyond replication initiation.

The minichromosome maintenance (or MCM) protein family is composed of six related proteins that are conserved in all eukaryotes. They were first identified by genetic screens in yeast and subsequently analyzed in other experimental systems using molecular and biochemical methods. Early data led to the identification of MCMs as central players in the initiation of DNA replication. More recent studies have shown that MCM proteins also function in replication elongation, probably as a DNA helicase. This is consistent with structural analysis showing that the proteins interact together in a heterohexameric ring. However, MCMs are strikingly abundant and far exceed the stoichiometry of replication origins; they are widely distributed on unreplicated chromatin. Analysis of mcm mutant phenotypes and interactions with other factors have now implicated the MCM proteins in other chromosome transactions including damage response, transcription, and chromatin structure. These experiments indicate that the MCMs are central players in many aspects of genome stability.

Animals↗
PubMed2005

Differential control of glucocorticoid receptor hormone-binding function by tetratricopeptide repeat (TPR) proteins and the immunosuppressive ligand FK506.

Many laboratories have documented the existence of tetratricopeptide repeat (TPR) proteins (also known as immunophilins) in hormone-free steroid receptor complexes. Yet, the distinct roles of these proteins in steroid receptor action are poorly understood. In this work, we have investigated the effects of four TPR proteins (FKBP52, FKBP51, Cyp40, and PP5) on hormone-binding function of glucocorticoid receptor (GR) endogenously expressed in mammalian L929 cells. As a first step, we treated L929 cells with select immunophilin ligands [FK506, rapamycin, cyclosporin A (CsA), and cyclosporin H (CsH)], which are commonly thought to increase the GR response to hormone by inhibiting membrane-based steroid exporters. As expected, all four immunophilin ligands increased both the intracellular concentration of dexamethasone and GR activity at the MMTV-CAT reporter. To determine whether these ligands could target GR function independent of steroid export mechanisms, we performed GR reporter gene assays under conditions of immunophilin ligand and dexamethasone treatment that yielded equal intracellular hormone concentrations. FK506 was found to stimulate GR transactivity beyond the effect of this ligand on hormone retention. In contrast, CsA only affected the GR through upregulation of hormone retention. By Scatchard analysis, FK506 was found to increase GR hormone-binding affinity while decreasing total binding sites for hormone. This result correlated with loss of GR-associated FKBP51 and replacement with PP5. Interestingly, no GR-associated Cyp40 was found in these cells, consistent with the ability of CsA ligand to only affect GR through the hormone export mechanism. To test the role of FKBP52 independent of FK506, FKBP52 was placed under the control of a tetracycline-inducible promoter. Upregulation of FKBP52 caused an increase in both GR hormone-binding affinity and transactivity, even in the absence of FK506. These results show that immunosuppressive ligands can alter GR hormone-binding function by changing the TPR protein composition of receptor complexes and that TPR proteins exert a hierarchical effect on this GR function in the following order: FKBP52 > PP5 > FKBP51.

Animals↗
PubMed2001

Statistical analysis of marginal count failure data.

Manufacturers want to assess the quality and reliability of their products. Specifically, they want to know the exact number of failures from the sales transacted during a particular month. Information available today is sometimes incomplete as many companies analyze their failure data simply comparing sales for a total month from a particular department with the total number of claims registered for that given month. This information--called marginal count data--is, thus, incomplete as it does not give the exact number of failures of the specific products that were sold in a particular month. In this paper we discuss nonparametric estimation of the mean numbers of failures for repairable products and the failure probabilities for nonrepairable products. We present a nonhomogeneous Poisson process model for repairable products and a multinomial model and its Poisson approximation for nonrepairable products. A numerical example is given and a simulation is carried out to evaluate the proposed methods of estimating failure probabilities under a number of possible situations.

Algorithms↗
PubMed2004

Staf50 is a novel p53 target gene conferring reduced clonogenic growth of leukemic U-937 cells.

The tumor suppressor gene p53 is a transcription factor that mediates both cell cycle arrest and apoptosis. Interestingly, p53 also induces differentiation of a number of tissues, including leukemic cells. However, although p53-mediated differentiation of leukemic U-937 cells depends on the transcriptional activity of p53, a p53 target gene mediating differentiation has hitherto not been identified. To screen for novel p53 target genes in leukemic cells, a cDNA microarray analysis was performed with U-937-4/ptsp53 cells, expressing a temperature-sensitive p53 mutant. We report that transcription of the Staf50 (stimulated transacting factor of 50 kDa) gene is upregulated in response to wild-type p53 in U-937-4, K562 and MCF-7 cells. Staf50 was directly activated by p53, as determined by the independence of de novo protein synthesis. Moreover, while the proximal promoter of Staf50 was found not to be p53 responsive, a functional enhancer-like p53-response element in intron 1 of the Staf50 gene was identified that was also transactivated by the p53-family member p73. Direct binding of p53 to the response element was shown by electrophoretic mobility shift analysis. Ectopic expression of Staf50 in U-937 cells resulted in reduced clonogenic growth. Moreover, levels of endogenous Staf50 mRNA correlated to all-trans retinoic acid-induced differentiation of promyelocytic NB-4 and HL60 cells, suggesting that Staf50 could be involved in proliferation and/or differentiation of leukemic cells.

Amino Acid Motifs↗
PubMed2002

Transformational leadership and moral reasoning.

Terms such as moral and ethical leadership are used widely in theory, yet little systematic research has related a sociomoral dimension to leadership in organizations. This study investigated whether managers' moral reasoning (n = 132) was associated with the transformational and transactional leadership behaviors they exhibited as perceived by their subordinates (n = 407). Managers completed the Defining Issues Test (J. R. Rest, 1990), whereas their subordinates completed the Multifactor Leadership Questionnaire (B. M. Bass & B. J. Avolio, 1995). Analysis of covariance indicated that managers scoring in the highest group of the moral-reasoning distribution exhibited more transformational leadership behaviors than leaders scoring in the lowest group. As expected, there was no relationship between moral-reasoning group and transactional leadership behaviors. Implications for leadership development are discussed.

Adult↗
PubMed1991

Using decision support systems to market prepaid medical plans.

A decision support system (DSS) is described that enables executives quickly and easily to (1) manage relevant marketing information and (2) convert the information into market-effective health plans. Because the system primarily uses available records and routine transactions, it can be implemented cost efficiently.

Cost-Benefit Analysis↗
PubMed1977

The international transfer of medical technology--an analysis and a proposal for effective monitoring.

The international transfer of medical technology to the developing countries occurs at four levels--medical education, research, and missions; multinational corporate transactions; technical assistance projects sponsored by the World Health Organization; and bilateral foreign aid programs. In this article, a proposal is made for effective monitoring of international medical technology transfer through political and legal means, including a specific code of conduct for corporations engaged in medical technology transfer. The development of "intermediate health technologies" along the lines suggested by E. F. Schumacher, and the advantages of such an innovation in terms of population issues and economic development are also discussed.

Communicable Disease Control↗
PubMed1994

Sexual behaviour in Kenya: implications for sexually transmitted disease transmission and control.

Sexual behaviour in Kenya in relation to STD transmission was investigated with a view to forming a basis for the more rational design of STD/HIV control interventions. Questionnaires were administered to a sample of 762 men and women attending eight health facilities in two urban centres. Equal numbers of STD patients (cases) and non-STD related clinic attenders (clinic controls) were selected, matched by gender and clinic. Another sample of 427 men and women was obtained from a random sampling of households in a slum area in Nairobi (community controls). Male STD patients who were unmarried, or married but living apart from their wives, reported a higher mean number of sex partners in the previous three months than did male clinic or community controls. Unmarried female STD patients reported a higher mean number of sex partners in the previous three months than did unmarried female clinic or community controls. Both male and female STD patients were more likely to report having been involved in commercial sex transactions in the previous three months than clinic or community controls. Considerable heterogeneity in sexual behaviour was apparent. In multivariate analysis, the most important predictor of STD acquisition for both men and women was the number of reported sex partners in the previous three months. In addition, for men only, marital status (unmarried, or married but living apart from their wives) and purchasing sex were significant predictors of being an STD patient. These data confirm the importance of commercial sex in STD transmission, and suggest that men play a bridging role between female sex workers and the general population of women.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1978

A strategy for research on psychological and social factors in hypertension.

Stress-related research has thus far failed to provide an adequate understanding of hypertension and other psychosomatic ailments for three main reasons: First, there has been a continual failure to view stress as a relational phenomenon, that is, as a particular kind of transaction between person and environment. Second, there has been much confusion about the social, psychological and physiological levels of stress analysis; each is to some extent independent of the other, so that what happens at one level cannot stand for what happens at another. Third, the predominant research model has been structural and static. That is, the researcher looks at some environmental or personality characteristic, treating it as a stable property, and attempts to relate it to the risk of hypertension across persons or groups. Such an approach overlooks the key social, psychological and physiological mediating processes (e.g., social supports, cognitive appraisals, and coping) that are concurrent with and have causal significance in blood pressure elevation or change. Structural research models need to be supplemented with process-oriented ones in which the same persons are observed across various adaptational encounters and over time.

Female↗
PubMed1995

Mergers and acquisitions: director and consultant liability exposure.

Corporate directors and their consultants must make decisions in an uncertain and changing health care environment. The losses each may face as a result of an incomplete analysis of the true value of the entities involved in mergers or acquisitions may extend beyond the failure of the transaction to the creation of personal liability as well. Accordingly, objective, careful, detailed, and fair decision-making based upon adequate information is more critical than ever for directors if they are to be able to take advantage of the business judgment rule, and also for consultants to avoid their own liability when transactions fail to deliver the values they have estimated.

Consultants↗
PubMedDate not supplied

Impact of manual and computer-assisted PACS for automated PACS.

Considerations of totally electronic picture archiving system (PACS) often neglect the fact that every radiology practice currently has some system for storing and retrieving images and related alphanumeric data. Although these systems are usually manual, many departments now use on-line computers to help manage film flow. In either event, the creation of electronic PACS can be viewed as a classic data processing problem of automating an existing system, and the conversion should proceed through the usual steps of documenting the existing system in detail, and conducting feasibility studies and cost-benefit analyses. Documenting current systems should be facilitated by computer-assisted PACS--particularly documenting transaction volumes which can be provided as a by-product of radiology information management systems. Similarly cost-benefit analysis should be facilitated, although the cost/benefit ratio may be less favorable when comparing automated to computer-assisted PACS. Finally, information management features such as those provided by current on-line radiology systems provide a framework necessary to realize the full benefits of automated PACS.

Computers↗
PubMedDate not supplied

Financial strategy considerations in the acquisition of biomedical instrumentation.

Equipment financing by conditional sale, lease-purchase, and true lease transactions is discussed with emphasis on equity and third-party reimbursement. Use of available funds, bank loans, and leasing are compared on a quantitative basis with an incremental cash flow (ICF) analysis. The ICF analysis involves cash accrual and present value techniques. It is demonstrated in a parametric study of lease rates, equipment useful-lives, and reimbursement levels that leasing is the preferred financing route when there is an expectation of obsolescence and when lease term maturities are significantly less than equipment useful-life. Once the decision has been made to lease equipment, it is important to structure the transaction to achieve the optimum mix of rate, term, buy-out options, and available tax shelter. Specific lease situations are given to illustrate the problems that should be considered by the lessee. Lease terms and conditions tailored to the requirements of the users of biomedical instrumentation are discussed in detail. Lessor selection and the importance of a professional working relationship between lessee and lessor also are explored. The capabilities of the lessor (financial, technical, and administrative) can be tapped by the hospital/clinic staff to develop significant advantages in addition to the financing.

Costs and Cost Analysis↗