Global changes in gene expression observed at the transition from growth to stationary phase in Listeria monocytogenes ScottA batch culture

Listeria monocytogenes is a food-borne Gram-positive bacterium that is responsible for a variety of infections (worldwide) annually. The organism is able to survive a variety of environmental conditions and stresses, however, the mechanisms by which L. monocytogenes adapts to environmental change are yet to be fully elucidated. An understanding of the mechanism(s) by which L. monocytogenes survives unfavourable environmental conditions will aid in developing new food processing methods to control the organism in foodstuffs. We have utilized a proteomic approach to investigate the response of L. monocytogenes batch cultures to the transition from exponential to stationary growth phase. Proteomic analysis showed that batch cultures of L. monocytogenes perceived stress and began preparations for stationary phase much earlier (approximately A(600) = 0.75, mid-exponential) than predicted by growth characteristics alone. Global analysis of the proteome revealed that the expression levels of more than 50% of all proteins observed changed significantly over a 7-9 h period during this transition phase. We have highlighted ten proteins in particular whose expression levels appear to be important in the early onset of the stationary phase. The significance of these findings in terms of functionality and the mechanistic picture are discussed.

The impact of organizational culture and reshaping capabilities on change implementation success: The mediating role of readiness for change

It was hypothesized that employees' perceptions of an organizational culture strong in human relations values and open systems values would be associated with heightened levels of readiness for change which, in turn, would be predictive of change implementation success. Similarly, it was predicted that reshaping capabilities would lead to change implementation success, via its effects on employees' perceptions of readiness for change. Using a temporal research design, these propositions were tested for 67 employees working in a state government department who were about to undergo the implementation of a new end-user computing system in their workplace. Change implementation success was operationalized as user satisfaction and system usage. There was evidence to suggest that employees who perceived strong human relations values in their division at Time 1 reported higher levels of readiness for change at pre-implementation which, in turn, predicted system usage at Time 2. In addition, readiness for change mediated the relationship between reshaping capabilities and system usage. Analyses also revealed that pre-implementation levels of readiness for change exerted a positive main effect on employees' satisfaction with the system's accuracy, user friendliness, and formatting functions at post-implementation. These findings are discussed in terms of their theoretical contribution to the readiness for change literature, and in relation to the practical importance of developing positive change attitudes among employees if change initiatives are to be successful.

Interaction of peptides, peptidomimetics and other drug candidates with the DI/tripeptide transport system in intestinal epithelial cells, using the in vitro caco-2 cell culture system

An uptake system was developed using Caco-2 cell monolayers and the dipeptide, glycyl-[3H]L-proline, as a probe compound. Glycyl-[3H]L-proline uptake was via the di-/tripeptide transport system (DTS) and, exhibited concentration-, pH- and temperature-dependency. Dipeptides inhibited uptake of the probe, and the design of the system allowed competitors to be ranked against one another with respect to affinity for the transporter. The structural features required to ensure or increase interaction with the DTS were defined by studying the effect of a series of glycyl-L-proline and angiotensin-converting enzyme (ACE)-inhibitor (SQ-29852) analogues on the uptake of the probe. The SQ-29852 structure was divided into six domains (A-F) and competitors were grouped into series depending on structural variations within specific regions. Domain A was found to prefer a hydrophobic function, such as a phenyl group, and was intolerant to positive charges and H+ -acceptors and donors. SQ-29852 analogues were more tolerant of substitutions in the C domain, compared to glycyl-L-proline analogues, suggesting that interactions along the length of the SQ-29852 molecule may override the effects of substitutions in the C domain. SQ-29852 analogues showed a preference for a positive function, such as an amine group in this region, but dipeptide structures favoured an uncharged substitution. Lipophilic substituents in domain D increased affinity of SQ-29852 analogues with the DTS. A similar effect was observed for ACE-NEP inhibitor analogues. Domain E, corresponding to the carboxyl group was found to be tolerant of esterification for SQ-29852 analogues but not for dipeptides. Structural features which may increase interaction for one series of compounds, may not have the same effect for another series, indicating that the presence of multiple recognition sites on a molecule may override the deleterious effect of anyone change. Modifying current, poorly absorbed peptidomimetic structures to fit the proposed hypothetical model may improve oral bioavailability by increasing affinity for the DTS. The stereochemical preference of the transporter was explored using four series of compounds (SQ-29852, lysylproline, alanylproline and alanylalanine enantiomers). The L, L stereochemistry was the preferred conformation for all four series, agreeing with previous studies. However, D, D enantiomers were shown in some cases to be substrates for the DTS, although exhibiting a lower affinity than their L, L counterparts. All the ACE-inhibitors and β-lactam antibiotics investigated, produced a degree of inhibition of the probe, and thus show some affinity for the DTS. This contrasts with previous reports that found several ACE inhibitors to be absorbed via a passive process, thus suggesting that compounds are capable of binding to the transporter site and inhibiting the probe without being translocated into the cell. This was also shown to be the case for oligodeoxynucleotide conjugated to a lipophilic group (vitamin E), and highlights the possibility that other orally administered drug candidates may exert non-specific effects on the DTS and possibly have a nutritional impact. Molecular modelling of selected ACE-NEP inhibitors revealed that the three carbonyl functions can be oriented in a similar direction, and this conformation was found to exist in a local energy-minimised state, indicating that the carbonyls may possibly be involved in hydrogen-bond formation with the binding site of the DTS.

Mediators of gram-positive septic shock

Septic shock can occur as a result of Gram-negative or Gram-positive infection and involves a complex interaction between bacterial factors and the host immune system producing a systemic inflammatory state that may progress to multiple organ failure and death. Gram-positive bacteria are increasingly becoming more prevalent especially Staphylococcus epidermidis in association with indwelling devices. Lipopolysaccaride (LPS) is the key Gram-negative component involved in this process, but it is not clear which components of Gram-positive bacteria are responsible for progression of this often fatal disease. The aim of this thesis was to investigate the effect of bacterial components on the immune systems. Lipid S, a short chain form of lipoteichoic acid (LTA) found to be excreted from bacteria during growth in culture medium was examined along with other Gram-positive cell wall components: LTA, peptidoglycan (PG) and wall teichoic acids (WTA) and LPS from Gram-negative bacteria. Lipid S, LTA, PG and LPS but not WTA all stimulated murine macrophages and cell lines to produce significant amounts of NO, TNF-a, IL-6 and IL-1 and would induce fever and tissue damage seen in inflammatory diseases. Lipid S proved to be the most potent out of the Gram-positive samples tested. IgG antibodies in patients serum were found to bind to and cross react with lipid S and LTA. Anti-inflammatory antibiotics, platelet activating factor (PAF), PAF receptor antagonists and monoclonal antibodies (mAbs) directed to LTA, CD14 and toll-like receptors were utilised to modulate cytokine and NO production. In cell culture the anti-LTA and the anti-CD14 mAbs failed to markedly attenuate the production of NO, TNF-a, IL-6 or IL-1, the anti-TLR4 antibody did greatly inhibit the ability of LPS to stimulate cytokine production but not lipid S. The tetracyclines proved to be the most effective compounds, many were active at low concentrations and showed efficacy to inhibit both lipid S and LPS stimulated macrophages to produce NO.

The 'Healthy Passport' intervention with older people in an English urban environment:effects of incentives and peer-group organisers in promoting healthy living

This paper reports the evaluation of the effectiveness of incentives (viz. points and prizes) and of peer-group organisers ('older people's champions') in the outcomes of a health-improvement programme for people aged 50 + years in a multi-ethnic district of the West Midlands, England. Health promotion activities Were provided, and adherence, outcome variables and barriers to adherence were assessed over six months, using a `passport' format. Those aged in the fifties and of Asian origin Were under represented, but people of Afro-Caribbean origin were well represented and proportionately most likely to stay in the project. Those of greater age and With more illness were most likely to drop out. There were significant improvements in exercise, diet and the uptake of influenza vaccines and eyesight tests, but slighter improvements in wellbeing. Positive outcomes related to the incentives and to liking the format. The number of reported barriers was associated with lower involvement and lack of change, as was finding activities too difficult, the level of understanding, and transport and mobility problems, but when these were controlled, age did not predict involvement. Enjoying the scheme was related to positive changes, and this was associated with support from the older people's champions.

Paracellular permeability is increased by basal lipopolysaccharide in a primary culture of colonic epithelial cells; an effect prevented by an activator of Toll-like receptor-2

Lipopolysaccharide (LPS), which generally activates Toll-like receptor 4 (TLR4), is expressed on commensal colonic bacteria. In a number of tissues, LPS can act directly on epithelial cells to increase paracellular permeability. Such an effect in the colon would have an important impact on the understanding of normal homeostasis and of pathology. Our aim was to use a novel primary culture of colonic epithelial cells grown on Transwells to investigate whether LPS, or Pam(3)CSK( 4), an activator of TLR2, affected paracellular permeability. Consequently, [(14)C]-mannitol transfer and transepithelial electrical resistance (TEER) were measured. The preparation consisted primarily of cytokeratin-18 positive epithelial cells that produced superoxide, stained for mucus with periodic acid-Schiff reagent, exhibited alkaline phosphatase activity and expressed TLR2 and TLR4. Tight junctions and desmosomes were visible by transmission electron microscopy. Basally, but not apically, applied LPS from Escherichia coli increased the permeability to mannitol and to a 10-kDa dextran, and reduced TEER. The LPS from Helicobacter pylori increased paracellular permeability of gastric cells when applied either apically or basally, in contrast to colon cells, where this LPS was active only from the basal aspect. A pan-caspase inhibitor prevented the increase in caspase activity caused by basal E. coli LPS, and reduced the effects of LPS on paracellular permeability. Synthetic Pam(3)CSK(4) in the basal compartment prevented all effects of basal E. coli LPS. In conclusion, LPS applied to the base of the colonic epithelial cells increased paracellular permeability by a mechanism involving caspase activation, suggesting a process by which perturbation of the gut barrier could be exacerbated. Moreover, activation of TLR2 ameliorated such effects.

Peer mentoring in higher education:a multidimensional approach to conceptualising the role of peer mentoring programmes in enhancing the first year undergraduate experience

Across the literature researchers agree that the concept of mentoring results in positive outcomes for both mentors and mentees alike (Enrich et al, 2004). From a pedagogical perspective, student focused mentoring activities in Higher Education are generally perceived to comprise dyadic or triadic relationships that encapsulate a diverse range of learning strategies and/or support mechanisms. Whilst there exists a significant amount of literature regarding the wider value of Peer Mentoring in Higher Education, there remains a notable gap in knowledge about the value of such programmes in enhancing the first year undergraduate experience and thus promoting a smooth transition to University. Using the emergent study findings of a large international project, a multidimensional conceptual framework bringing together the theoretical, conceptual and contextual determinants of Peer Mentoring is proposed. This framework makes a distinctive contribution to current pedagogical theory and practice – particularly in relation to the first year experience.

Development of a neurotoxicity test-system, using human post-mitotic, astrocytic and neuronal cell lines in co-culture

Astrocytes are essential for neuronal function and survival, so both cell types were included in a human neurotoxicity test-system to assess the protective effects of astrocytes on neurons, compared with a culture of neurons alone. The human NT2.D1 cell line was differentiated to form either a co-culture of post-mitotic NT2.N neuronal (TUJ1, NF68 and NSE positive) and NT2.A astrocytic (GFAP positive) cells (∼2:1 NT2.A:NT2.N), or an NT2.N mono-culture. Cultures were exposed to human toxins, for 4 h at sub-cytotoxic concentrations, in order to compare levels of compromised cell function and thus evidence of an astrocytic protective effect. Functional endpoints examined included assays for cellular energy (ATP) and glutathione (GSH) levels, generation of hydrogen peroxide (H2O2) and caspase-3 activation. Generally, the NT2.N/A co-culture was more resistant to toxicity, maintaining superior ATP and GSH levels and sustaining smaller significant increases in H2O2 levels compared with neurons alone. However, the pure neuronal culture showed a significantly lower level of caspase activation. These data suggest that besides their support for neurons through maintenance of ATP and GSH and control of H2O2 levels, following exposure to some substances, astrocytes may promote an apoptotic mode of cell death. Thus, it appears the use of astrocytes in an in vitro predictive neurotoxicity test-system may be more relevant to human CNS structure and function than neuronal cells alone. © 2007 Elsevier Ltd. All rights reserved.

Are guanxi-type supervisor-subordinate relationships culture-general? An eight-nation test of measurement invariance

Three dimensions of subordinate-supervisor relations (affective attachment, deference to supervisor, and personal-life inclusion) that had been found by Y. Chen, Friedman, Yu, Fang, and Lu to be characteristic of a guanxi relationship between subordinates and their supervisors in China were surveyed in Taiwan, Singapore, and six non-Chinese cultural contexts. The Affective Attachment and Deference subscales demonstrated full metric invariance whereas the Personal-Life Inclusion subscale was found to have partial metric invariance across all eight samples. Structural equation modeling revealed that the affective attachment dimension had a cross-nationally invariant positive relationship to affective organizational commitment and a negative relationship to turnover intention. The deference to the supervisor dimension had invariant positive relationships with both affective and normative organizational commitment. The personal-life inclusion dimension was unrelated to all outcomes. These results indicate the relevance of aspects of guanxi to superior-subordinate relations in non-Chinese cultures. Studies of indigenous concepts can contribute to a broader understanding of organizational behavior. © The Author(s) 2014.

Apoptosis induction in gastric mucous cells in vitro:lesser potency of Helicobacter pylori than Escherichia coli lipopolysaccharide, but positive interaction with ibuprofen

Non-steroidal anti-inflammatory drugs (NSAIDs) cause peptic ulcer disease, but whether they interact with Helicobacter pylori to promote damage is controversial. Moreover, the reported induction of apoptosis in gastric cells by H. pylori lipopolysaccharide (LPS) (10-9 g /ml) contrasts with studies showing low immunological potency of this LPS. Therefore, the effects of LPS from H. pylori NCTC 11637 and Escherichia coli 0111:B4 on apoptosis in a primary culture of guinea-pig gastric mucous cells were investigated in the presence and absence of the NSAID, ibuprofen. Cell loss was estimated by a crystal violet assay, and apoptosis determined from caspase activity and from condensation and fragmentation of nuclei. Exposure to E. coli LPS for 24 h caused cell loss and enhanced apoptotic activity at concentrations ≥ 10-9 g/ml, but similar effects were only obtained with H. pylori LPS at concentrations ≥10-6 g/ml. Although ibuprofen (250 μM) caused cell loss and apoptosis, addition of either E. coli or H. pylori LPSs further enhanced these effects. In conclusion, LPS and ibuprofen interact to enhance gastric cell loss and apoptosis. In such interactions, E. coli LPS is more potent than that of H. pylori. The low potency of H. pylori LPS may contribute to a chronic low-grade gastritis that can be enhanced by the use of NSAIDs. © W. S. Maney & Son Ltd.

Tackling transition:the value of peer mentoring

This paper is aimed at those interested in the promotion of student retention in higher education; particularly those with an interest in peer mentoring as a means of student support. It critically discusses the results of an exploratory study analysing the perceptions of peer mentors and mentees within five universities in the United Kingdom. The aim of the study was to analyse how student peer mentoring can aid transition into university by focusing specifically on how senior students can support their junior counterparts in their first year at university. The paper discusses the results of a survey which was completed by 329 student peer mentors and mentees. Focusing on the benefits and outcomes of participation in Mentoring Programmes, the survey was distinctive in that it asked mentors and mentees similar questions. From a theoretical perspective, the paper contributes to debates about peer support in higher education showing that participation in such programmes can have positive outcomes from both social and pedagogic perspectives. Practically speaking, the results have important implications for Higher Education Institutions as the research highlights the importance of putting into place formally structured Peer Mentoring Programmes which facilitate student support at a time when new students are most at risk of ‘dropping out’.

The influence of culture and market orientation on services brands:insights from Irish banking and retail firms

Purpose: This paper aims to examine the influence of the culture of the service firm on its interpretation of the role of the brand and on the development and implementation of its brand values. Design/methodology/approach: A grounded theory approach was used. Interviews were conducted with 20 managers within two leading banking firms in Ireland and two leading grocery retailers in Ireland. Findings: The development of the brand, and its role within the firm, is closely related to the firm's culture. The research shows obstacles and opportunities created by the cultural context of firms wishing to disseminate and embed a set of brand values. The paper presents an "involvement model" of brand values implementation and outlines changes required to implement brand values. Research limitations/implications: The study was bound by access to firms, and managers' availability. The authors sought an insight into the relationship between each firm's culture and its brands. They advocate quantitative research to further investigate the findings within these service sectors and to test proposed antecedents (transformational leadership, employee involvement) and outcomes (employee-based brand equity and consumer-based brand equity) of values adoption. Practical implications: The paper identifies aspects of retail and banking cultures which support or detract from brand development. In particular, it presents the learnings from successful brand values implementation in a clan culture, aspects of which are applicable across other cultures. Originality/value: The paper provides valuable insights into the role of the brand within the service firm and the positive and negative influence of context on brand values and their development and implementation. © Emerald Group Publishing Limited.

Effects of special educator peer coaching on general educator effectiveness in inclusive classroom settings

This dissertation investigated the effects of a peer coaching relationship between a special education teacher and two general education teachers. More specifically, a two-tier multiple baseline design across subjects was used to evaluate the effects of peer coaching on the general education teachers' use of effective instructional practices (EIPs) and subsequent effects on the engagement rate and academic performance of students with and without disabilities. The peer coaching process included modeling, direct support, and feedback on the use of effective instructional practices including getting student attention, giving specific directions, asking specific questions with wait time, contingent positive reinforcement, positive error correction, precorrection, prompting, and proximity control. A 30-second partial interval recording procedure was used to observe the general education teachers' use of effective instructional practices and student engagement rates. Student participants' academic performance was measured using weekly quizzes. ^ Peer coaching resulted in an overall increase in the teachers' use of EIPs. One general education teacher had a 30% increase in average EIP use from 46% during the baseline phase to 76% during intervention. Student engagement for her two student participants with and without disabilities indicated an increase from 54% to 69% and from 47% to 65% respectively. Results for the second general education teacher indicated a 34% increase in average EIP use from 55% during the baseline to 89% during intervention. Student engagement for the two student participants with and without disabilities in her class increased from 48% to 83% and from 29% to 71% respectively. Student academic performance showed a small increase. In follow-up observations, the effects of peer coaching on teacher use of EIPs and student engagement and academic performance were maintained. ^ The results of this study suggest that using peer coaching to support general education teachers can be an effective method to improve the educational outcomes of students with and without disabilities in general education. Further research is needed to investigate the effects of peer coaching with other special and general educator partnerships and other student participants. ^

The effect of a reform on a school organizational culture after receiving a failing Florida Comprehensive Assessment Test rating

The purpose of this research was to explore the effects of a reform that took place in an elementary school during 2000/2001 as a result of a failure rating on the Florida Comprehensive Assessment Test on the structure and the personnel of the organization. ^ The exploration took place over a period of 10 months starting in August 2000 until June 2001. It focused on the effect of the failure rating on the: (a) structure and operation of the school; (b) morale, beliefs, behaviors, and daily lives of teachers and the principal; and (c) the effect of the reform effort on the leadership style of the principal, whether she became a transactional or a transformative leader. ^ The researcher assumed the role of a participant observer. Data sources were her personal recollections of major events that took place during the year of the reform, interviews, observations, and school documents. The sample included 15 teachers present during the time of the reform. Ten taught second through fifth grade. The remaining five participants were the music teacher, the counselor, and the writing, reading and technology specialists. Together they represented the instructional team or represented special education areas. ^ The findings indicated that the reform effort had an effect on the structure and the operation of the school. The changes included reorganization of the physical set up, changes in curriculum and instruction, changes in the means of communication among the staff, and the addition of new staff members including an official agent of change. The reform had a greater effect on the daily lives of teachers and their morale than on their beliefs and behaviors. Teachers reported that during the effort their daily lives were stressful and their morale very low due to the enormous expectations that they had to meet. On the other hand, the reform effort had a positive effect on the daily life, morale, beliefs, and behaviors of the principal. It energized her. She spoke positively about the change. She functioned as an effective, positive, resilient transactional leader who did what was necessary in order to enable the teachers to cope with the complex situation. ^