A novel assay for drug-DNA binding mode, affinity, and exclusion number: scanning force microscopy.

Determining the mode-of-binding of a DNA ligand is not always straightforward. Here, we establish a scanning force microscopic assay for mode-of-binding that is (i) direct: lengths of individual DNA-ligand complexes are directly measured; (ii) rapid: there are no requirements for staining or elaborate sample preparation; and (iii) unambiguous: an observed increase in DNA length upon addition of a ligand is definitive evidence for an intercalative mode-of-binding. Mode-of-binding, binding affinity, and site-exclusion number are readily determined from scanning force microscopy measurements of the changes in length of individual drug-DNA complexes as a function of drug concentration. With this assay, we resolve the ambiguity surrounding the mode of binding of 2,5-bis(4-amidinophenyl) furan (APF) to DNA and show that it binds to DNA by nonintercalative modes. APF is a member of an important class of aromatic dicationic drugs that show significant activity in the treatment of Pneumocystis carinii pneumonia, an opportunistic infection that is the leading cause of death in AIDS patients.

A chromosomal locus required for copper resistance, competitive fitness, and cytochrome c biogenesis in Pseudomonas fluorescens.

A chromosomal locus required for copper resistance and competitive fitness was cloned from a strain of Pseudomonas fluorescens isolated from copper-contaminated agricultural soil. Sequence analysis of this locus revealed six open reading frames with homology to genes involved in cytochrome c biogenesis in other bacteria, helC, cycJ, cycK, tipB, cycL, and cycH, with the closest similarity being to the aeg-46.5(yej) region of the Escherichia coli chromosome. The proposed functions of these genes in other bacteria include the binding, transport, and coupling of heme to apocytochrome c in the periplasm of these Gram-negative bacteria. Putative heme-binding motifs were present in the predicted products of cycK and cycL, and TipB contained a putative disulfide oxidoreductase active site proposed to maintain the heme-binding site of the apocytochrome in a reduced state for ligation of heme. Tn3-gus mutagenesis showed that expression of the genes was constitutive but enhanced by copper, and confirmed that the genes function both in copper resistance and production of active cytochrome c. However, two mutants in cycH were copper-sensitive and oxidase-positive, suggesting that the functions of these genes, rather than cytochrome c oxidase itself, were required for resistance to copper.

Function of the pre-T-cell receptor alpha chain in T-cell development and allelic exclusion at the T-cell receptor beta locus.

The pre-T-cell receptor, composed of the T-cell receptor (TCR) beta chain (TCRbeta), pre-Talpha (pTalpha) chain, and CD3 molecules, has been postulated to be a transducer of signals during the early stages of T-cell development. To examine the function of the transmembrane pTalpha chain during tbymocyte development, we generated pTalpha-/- embryonic stem cells and assayed their ability to differentiate into lymphoid cells in vivo after injection into recombination-activating gene (RAG)-2-deficient blastocysts. Thymocytes representing all stages of T-cell differentiation were detected in the thymus of pTalpha-/- chimeric mice, indicating that thymocyte development can occur without pTalpha. However, greatly reduced thymocyte numbers and substantially increased percentages of both CD4-CD8- thymocytes and TCRgammadelta+ thymocytes suggest that pTalpha plays a critical role in thymocyte expansion. To investigate the role of the pTalpha chain in allelic exclusion at the TCRbeta locus, a functionally rearranged TCRbeta minigene was introduced into pTalpha-/- and pTalpha+/- embryonic stem cells, which were subsequently assayed by RAG-2-deficient blastocyst complementation. In the absence of pTalpha, expression of the transgenic TCRbeta inhibited rearrangement of the endogenous TCRbeta locus to an extent similar to that seen in normal TCRbeta transgenic mice, suggesting that pTalpha may not be required for signaling allelic exclusion at the TCRbeta locus.

Significant competitive advantage conferred by meiosis and syngamy in the yeast Saccharomyces cerevisiae.

The presumed advantages of genetic recombinations are difficult to demonstrate directly. To investigate the effects of recombination and background heterozygosity on competitive ability, we have performed serial-transfer competition experiments between isogenic sexual and asexual strains of the yeast Saccharomyces cerevisiae. The members of these diploid pairs of strains differed only in being heterozygous (sexual) or homozygous (asexual) at the mating type or MAT locus. Competing pairs had either a completely homozygous or a heterozygous genetic background, the latter being heterozygous at many different loci throughout the genome. A round of meiotic recombination (automixis) conferred a large and statistically significant enhancement of competitive ability on sexual strains with a heterozygous genetic background. By contrast, in homozygous background competitions, meiosis decreased the sexual strains' initial relative competitive ability. In all cases, however, the sexual strains outcompeted their isogenic asexual counterparts, whether meiotic recombination had occurred or not. In some genetic backgrounds, this was due in part to an overdominance effect on competitive advantage of heterozygosity at the MAT locus. The advantage of the sexual strains also increased significantly during the course of the homozygous background competitions, particularly when meiosis had occurred. This latter effect either did not occur or was very weak in heterozygous background competitions. Overall, sexual strains with heterozygous genetic backgrounds had a significantly higher initial relative competitive ability than those with homozygous backgrounds. The advantage of mating type heterozygosity in this organism extends far beyond the ability to recombine meiotically.

A single amino acid in gamma-aminobutyric acid rho 1 receptors affects competitive and noncompetitive components of picrotoxin inhibition.

A class of bicuculline-insensitive gamma-aminobutyric acid (GABA) receptors, GABAC, has been identified in retina. Several lines of evidence indicate that GABAC receptors are formed partially or wholly of GABA rho subunits. These receptors generate a Cl- current in response to GABA but differ from GABAA receptors in a number of ways. Picrotoxin, widely accepted as a noncompetitive antagonist of GABAA receptors, displays competitive and noncompetitive antagonism of GABAC receptors in perch and bovine retina and GABA rho 1 receptors expressed in Xenopus oocytes. The aim of this study was to identify the molecular basis of the two components of picrotoxin inhibition of GABA rho 1 receptors. By using a domain-swapping and mutagenesis strategy, a difference in picrotoxin sensitivity between rho 1 and rho 2 receptors was localized to a single amino acid in the putative second transmembrane domain. Substitution of this amino acid with residues found in the analogous position in highly picrotoxin-sensitive glycine alpha and GABAA subunits increased the sensitivity of rho 1 mutants 10- to 500-fold. Importantly, the competitive component of picrotoxin inhibition of the rho 1 mutant receptors was almost eliminated. These findings demonstrate that an amino acid in the putative channel domain of GABA rho 1 receptors influences picrotoxin sensitivity and mediates agonist binding by an allosteric mechanism.

Potent, structurally constrained agonists and competitive antagonists of corticotropin-releasing factor.

Predictive methods, physicochemical measurements, and structure activity relationship studies suggest that corticotropin-releasing factor (CRF; corticoliberin), its family members, and competitive antagonists (resulting from N-terminal deletions) usually assume an alpha-helical conformation when interacting with the CRF receptor(s). To test this hypothesis further, we have scanned the whole sequence of the CRF antagonist [D-Phe12,Nle21,38]r/hCRF-(12-41) (r/hCRF, rat/human CRF; Nle, norleucine) with an i-(i + 3) bridge consisting of the Glu-Xaa-Xaa-Lys scaffold. We have found astressin [cyclo(30-33)[D-Phe12,Nle21,38,Glu30,Lys33]r/ hCRF(12-41)] to be approximately 30 times more potent than [D-Phe12,Nle21,38]r/hCRF-(12-41), our present standard, and 300 times more potent than the corresponding linear analog in an in vitro pituitary cell culture assay. Astressin has low affinity for the CRF binding protein and high affinity (Ki = 2 nM) for the cloned pituitary receptor. Radioiodinated [D-125I-Tyr12]astressin was found to be a reliable ligand for binding assays. In vivo, astressin is significantly more potent than any previously tested antagonist in reducing hypophyseal corticotropin (ACTH) secretion in stressed or adrenalectomized rats. The cyclo(30-33)[Ac-Pro4,D-Phe12,Nle21,38,Glu30,Lys33++ +]r/hCRF-(4-41) agonist and its linear analog are nearly equipotent, while the antagonist astressin and its linear form vary greatly in their potencies. This suggests that the lactam cyclization reinstates a structural constraint in the antagonists that is normally induced by the N terminus of the agonist.

Human Resource's Role in Leveraging Knowledge Management as a Competitive Advantage

The ultra-competitiveness in the twenty-first century's business environment suggests that, to survive, human resource (HR) must prove its value. The knowledge economy presents HR the opportunity to demonstrate its added value through leveraging knowledge management (KM) as a competitive advantage. This capstone reviews the literature on business strategy, knowledge, KM, KM success, and HR to reveal links between these subjects and illustrate three roles HR can undertake to leverage KM as a competitive advantage. The capstone also assesses HR's current contributions to KM through a survey of 42 HR professionals. Survey results suggest that, although HR contributes to the three KM roles, HR is not currently fully capitalizing on the opportunity to become a strategic business partner through KM.

Who performs a stronger response to whom? Detecting individual competitive actions and reactions

This study analyzes the degree of competition through individual actions and reactions. Empirical support for this analysis has derived mainly from structural econometric models describing the nature of competition. This analysis extends the existing literature by empirically considering a direct measurement of competition through the analysis of the competitive actions and responses, and describing how firms compete within and between strategic groups. We estimate the firms’ conduct in the Spanish deposits market with 146 firms and 18,888 observations. This is a specially compelling context for the banking industry, in which a deregulation process gives rise to the adoption of aggressive strategies seeking to increase the market shares of deposit accounts; thus, producing a turbulent situation of increasing rivalry. Our results offer a deeper understanding of the firms’ competitive behavior, since we identify different patterns of actions and reactions depending upon the strategic group the firm belongs to.

The effects of quality and environmental management on competitive advantage: A mixed methods study in the hotel industry

This paper reports results derived from a mixed methods study where 13 hotel managers were initially interviewed, followed by a quantitative study of 355 additional managers. Data were analysed using partial least squares path modelling. The research question related to the relationship between quality and environmental management and the competitive advantage sought by hotels. The results indicate that quality management and environmental management permit the improvement of competitive advantage in terms of both costs and differentiation. Moreover, hotels implementing quality programmes find fewer obstacles in implementing environmental management.

Organizational design, quality management and competitive advantage in hotels

Purpose – The aim of this study is to examine the relationship between practices of quality management (QM) and the characteristics of organizational design, and QM and competitive advantage. Design/methodology/approach – The study uses a partial least squares approach to test these relationships in 350 hotels in Spain. Findings – The findings show that QM influences specialization, formalization and interdepartmental interactions, and that QM practices influence both cost and differentiation competitive advantage. The results also indicate the importance of QM strategic and operational systems as practices that have a key impact on the characteristics of organizational design. Similarly, the QM operational system is key in the relationship between QM and cost competitive advantage. Finally, the QM operational, information and strategic systems positively influence differentiation competitive advantage. Practical implications – When hotels adopt QM practices, there will be significant changes in a number of organizational variables, including specialization, formalization and interdepartmental interactions. This paper provides empirical evidence that QM practices improve both cost and differentiation competitive advantage in the hotel industry. Originality/value – There has been little research on the effects of QM on organizational design in the hotel industry. The contribution of this paper is that analyze the effects of QM on organizational design and competitive advantage, extending knowledge about these issues in a specific sector.

An Evaluation of Personal and Contextual Factors in Competitive Youth Sport

Because millions of youth are involved in sport, the sport context is important to consider in advancing the growth experiences of young people (Cˆot´e et al., 2007; Fraser-Thomas et al., 2005). Furthermore, research in developmental psychology has highlighted the value of structured programs, including sport, in helping to promote positive youth development (Fredricks & Eccles, 2006). Youth sport involvement has been linked to high levels of enjoyment (Scanlan et al., 1989), however, negative outcomes, such as burnout, have also been reported (Gould et al., 1996). In the present study, the Developmental Assets Profile (Search Institute, 2004) was used to explore personal (internal assets) and contextual (external assets) outcomes associated with youth sport. Results suggest that three particular assets (positive identity, empowerment, and support) are important to focus on in youth sport programs to decrease burnout symptoms and enhance enjoyment. Path analyses were also conducted to test a proposed model and exploratory results confirmed links of particular assets to sport outcomes. The results are discussed in terms of integration with Bronfenbrenner’s ecological theory (1999) and recommendations are suggested for sport programmers to consider to develop these assets within youth sport.

Understanding Dropout and Prolonged Engagement in Adolescent Competitive Sport

Objectives: The purpose of this study is to gain understanding of training patterns and roles of significant others (i.e. coaches, parents, peers, and siblings) in adolescent swimmers’ sport participation patterns. Design: The developmental model of sport participation [Côté, J., Baker, J., & Abernethy, B. (2003). From play to practice: A developmental framework for the acquisition of expertise in team sport. In J. Starkes, & K. A. Ericsson (Eds.), Recent advances in research on sport expertise (pp. 89–114). Champaign, IL: Human Kinetics; Côté, J., & Fraser-Thomas, J. (2007). Youth involvement in sport. In P. R. E. Crocker (Ed.), Introduction to sport psychology: A Canadian perspective (pp. 266–294). Toronto: Pearson Prentice Hall] was used as a framework. Method: Ten dropout and 10 engaged swimmers, matched on key demographic variables participated in a semi-structured qualitative interview. Results: Groups had many similar experiences (e.g. early training, supportive and unsupportive coaches, involved parents). However, only dropouts spoke of early peak performances, limited one-on-one coaching, pressuring parents during adolescence, lack of swimming peers during adolescence, and sibling rivalries. In contrast, only engaged athletes spoke of clubs’ developmental philosophies, coaches’ and parents’ open communication, school friends’ support, and siblings’ general positive influences. Conclusions: Findings highlight the importance of appropriately structured programs and the fragility of athletes’ relationships with significant others during the adolescent years. Implications for sport programmers, coaches, and parents are discussed.

Experience in Competitive Youth Sport and Needs Satisfaction: The Singapore Story

The purpose of this study was to examine the relationship between sport experiences and psychological needs satisfaction of Singapore high school athletes who were involved in inter-school competition. A total of 1250 school athletes from 22 sports participated in the study. The athletes were between 13 and 18 years old and had an average of 3 years of experience in school sport (SD=.18). Cluster analysis was employed to identify homogenous groups based on the seven developmental experiences domains of the Youth Experience Survey (YES 2.0; Hansen & Larson, 2005). A one-way analysis of variance (ANOVA) was conducted to determine whether differences existed among the clusters in terms of psychological needs satisfaction (i.e., sense of autonomy, perceived competence and relatedness). The results of the cluster analysis showed that there were different subgroups of athletes with distinct developmental experiences, and they varied in the degree to which their psychological needs were satisfied. Generally, subgroups that had high levels of positive experiences and low levels of negative experiences in sport had better fulfillment of psychological needs. It is important to ensure that policies and programmes are formulated, delivered and monitored effectively to promote positive experiences for youth who are involved in competitive sports.