Switching the mode of metabolism in the yeast Saccharomyces cerevisiae

The biochemistry of most metabolic pathways is conserved from bacteria to humans, although the control mechanisms are adapted to the needs of each cell type. Oxygen depletion commonly controls the switch from respiration to fermentation. However, Saccharomyces cerevisiae also controls that switch in response to the external glucose level. We have generated an S. cerevisiae strain in which glucose uptake is dependent on a chimeric hexose transporter mediating reduced sugar uptake. This strain shows a fully respiratory metabolism also at high glucose levels as seen for aerobic organisms, and switches to fermentation only when oxygen is lacking. These observations illustrate that manipulating a single step can alter the mode of metabolism. The novel yeast strain is an excellent tool to study the mechanisms underlying glucose-induced signal transduction. © 2004 European Molecular Biology Organization.

Studies on the mode of action and beta-lactamase inhibitory properties of novel oxapenem compounds

Four novel oxapenem compounds were evaluated for their ß-lactamase inhibitory and antibacterial properties. Two (AM-112 and AM-113) displayed intrinsic antibacterial activity with MICs of between 2 to 16µg/ml and 0.5-2µg/ml against Escherichia coli and methicillin-sensitive and -resistant Staphylococcus aureus, respectively. The isomers of these compounds, AM-115 and AM-114 did not display significant antibacterial activity. Combination of the oxapenems with ceftazidime afforded protection against ß-lactamase-producing strains, including hyperproducers of class C enzymes and extended-spectrum ß-lactamase enzymes. A fixed 4µg/ml concentration of AM-112 protected a panel of eight cephalosporins against hydrolysis by class A and class C ß-lactamase producers. In vivo studies confirmed the protective effect of AM-112 for ceftazidime against ß-lactamase producing S. aureus, Enterobacter cloacae and E. coli strains in a murine intraperitoneal infection model. Each of the oxapenems inhibited class A, class C and class D ß-lactamases isolated from whole cells and purified by isoelectric focusing. AM-114 and AM-115 were as effective as clavulanic acid against class A enzymes. AM-112 and AM-113 were less potent against these enzymes. Class C and class D enzymes proved very susceptible to inhibition by the oxapenems. Molecular modelling of the oxapenems in the active site of the class A. TEM-1 and class C P99 enzymes identified a number of potential sites of interaction. The modelling suggested that Ser-130 in TEM-1 and Tyr-150 in P99 were likely candidates for cross-linking of the inhibitor, leading to inhibition of the enzyme. Morphology studies indicated that sub-inhibitory concentrations of the oxapenems caused the formation of round-shaped cells in E. coli DC0, indicating inhibition of penicillin-binding protein 2 (PBP2). The PBP affinity profile of AM-112 was examined in isolated cell membranes of E. coli DC0, S. aureus NCTC 6571, Enterococcus faecalis SFZ and E. faecalis ATCC 29213, in competition with a radiolabelled penicillin. PBP2 was identified as the primary target for AM-112 in E. coli DC0. Studies on S. aureus NCTC 6571 failed to identify a binding target. AM-112 bound to all the PBPs of both E. faecalis strains, and a concentration of 10µg/ml inhibited all the PBPs except PBP3.

The mode of action of a lipid mobilising factor in cancer cachexia

Cachexia is characterised by a progressive weight loss due to depletion of both skeletal muscle and adipose tissue. The loss of adipose tissue is due to the production of a tumour-derived lipid mobilising factor (LMF), which has been shown to directly induce lipolysis in isolated epididymal murine white adipocytes. The administration of LMF to a non-tumour bearing mice produced a rapid weight loss, with a specific reduction in carcass lipid with also some redistribution of lipid with the accumulation of lipid in the liver. There was also up-regulation of uncoupling protein-1 and -2 mRNA and protein expression in brown adipose tissue, suggesting that an adaptive process occurs due to increased energy mobilisation. There was also up-regulation of UCP-2 in the livers of LMF treated mice, suggesting a protective mechanism to the build up of lipid in the livers, which would produce free radical by-products. LMF was also shown to stimulate cyclic AMP production in CHO-K1 cells transfected with human -3 adrenergic receptors and inhibited by the -β3 antagonist SR59230A. LMF binding was also inhibited by SR59230A in isolated receptors. This suggests that LMF mediates its effects through a β3 adrenergic receptor. There were also changes in glucose and fatty acid uptake in LMF treated mice, which suggests metabolic changes are occurring. The study suggests that a tumour derived lipolytic factor acts through the 3 adrenoceptor producing effects on lipid mobilisation, energy expenditure and glucose metabolism.

The mode of action of bradykinin

The action of bradykinin on transepithelial transfer of sodium and water in isolated rat jejunum and on smooth muscle contraction of rat terminal ileum has been investigated. (1) Bradykinin was shown to stimulate transfer at low control transfer, inhibit transfer at high control transfer and have no effect at intermediate transfer in rat jejunal sacs. Stimulation of transfer occurred only when bradykinin was in the serosal solutiun while inhibition of transfer occurred whether bradykinin was in the aerosal or mucosal solution. Bradykinin-induced stimulation of transfer was not affected by adrenalectomy, nephrectomy, combined adrenalectomy-nephrectomy,  nor maintenance on 1% saline drinking solution or low sodium diet pretreatment. Meclofenamic acid abolished the bradykinin-induced inhibition of water transfer while prostaglandins A1, E1 aud F2α all potentiated this action. Theophylline inhibited water transfer and potentiated the bradykinin-induced inhibition of water transfer. Cyclic AMP and dibutyryl cyclic AMP both inhibited water transfer and the bradykinin-induced inhibition of water transfer was potentiated by the latter. ( 2 ) Bradykinin-induced contractions of rat terminal ileum were little affected by hyoscine while those of acetylcholine were abolished. Anoxia reduced markedly responses tv bradykinin while those of acetylcholine were little affected . Theophylline reduced the responses of rat terminal ileum to bradykinin significantly more than those to acetylcholine. Aspirin and indomethacin reduced markedly the responses to bradykinin while not affecting those to acetylcholine and PGT2. Meslofenamic acid at a concentration of 3.4 µM blocked bradykinin-induced contractions but had no effect on those to acctylcholine, PGE2 or PGF2 and at a concentration of 17. 0 µM drastically reduced bradykinin responses but also reduced those to acetylcholine, PGE2 and PGF2α• Flufenamic acid drastically reduced responses to bradykinin while not affecting those to acetylcholine and PGE2 and slightly affecting those to PGF2α. Polyphloretin phosphate reduced responses to bradykinin, PGF2α and PGE2 but not acetylcholine . Diphloretin phosphate reduced responses to bradykinin, PGF2 and PGE2 in a dose dependent manner but not those to acetylcholine. SC 19220 , in a dose dependent manner, inhibited responses to bradykinin and PGE2 but not to acetylcholine and PGF2. 7 oxa - 13 -prostynoic acid non specifically reduced responses to acetylcholine, bradykinin and PGE2. Bradykinin, in the presence of SQ 20881 , increased the release of prostaglandin-like activity from rat terminal ileum and this was reduced or abolished in the presence of indomethacin, aspirin, meclofenamic acid or flufenamio acid. The extract of PG-like activity did not appear as PGE, PGA or PGFon TLC, but included a substance with similar mobility as 15-Keto-prosta-glandin E2.

Studies on the mode of cytotoxicity of imidazotetraziones

The irnidazotetrazinones are a novel group of anti tumour agents which have demonstrated good activity against a range of murine tumours and human xenografts. They possess a structure activity relationship similar to the anti tumour triazenes, with the chloroethyl (mitozolomide) and methyl (temozolomide) analogues being active antitumour agents, whilst the ethyl (CCRG 82019) and higher homologues are inactive. This thesiS attempts to elucidate the biological mechanisms responsible for the strict structure-activity relationship observed amongst the imidazotetrazinones. Mitozolomide is the only agent chemically capable of cross-linking DNA , which has been suggested to be responsible fo r the cytotoxicity of this group of agents. Only mitozolomide and ternozolornide Exhibit a marked ditferential toxicity towards the 0 -alkylguanine-DNA alkyltransferase deficient GM892A (Mer-) cell line rather than the proficient Raji cell line (Mer+). The rate of uptake of imidazotetrazinones into cells is similar for all three agents in both cell lines, and does not explain the differing sensitivities to these agents. The effect of drug treatment on the incorporation of precursors into macromolecules, and their pool sizes, was examined. Temozolomide administration was found to alter de novo protein synthesis in both GM892A and Raji cells. Flow cytometric analysis revealed that temozolomide and CCRG 82019 block cells in late S/G2/M phase of the cell cycle , similar to that observed with mitozolomide. The extent of reaction of all three drugs with isolated macromolecules and cellular macromolecules was determined, and differences found, with cellular repair processes influencing the number of alkyl lesions remaining bound to macromolecules. The specific bases formed in calf thymus DNA after treatment with either temozolornide and CCRG 82019 was measured, and it was found that the types and relative amounts of lesions formed, differed, as well as the total level of alkylation. Whereas DNA extracted from imidazotetrazinone treated cells is not affected in its ability to support RNA polymerase activity, an effect is observed on the ability to extract DNA polymerase from drug treated cells. This may suggest that the alkylated DNA must be in intact chromatin for the lesion to manifest its effects. Temozolomide and methyl methanesulphonate do got appear to act with a synergistic mode of action. The 0 -position of guanine is suspected to be a critical site for the action of these types of drugs.

Maintenance of communication in primary classrooms : some evidence for the role of elicitation and code-switching in English medium schools in Kenya, with implications for teaching

This is a study of specific aspects of classroom interaction primary school level in Kenya. The study entailed the identification of the sources of particular communication problems during the change-over period from Kiswahili to English medium teaching in two primary schools. There was subsequently an examination of the language resources which were employed by teachers to maintain pupil participation in communication in the light of the occurrence of possibility of occurrence of specific communication problems. The language resources which were found to be significant in this regard concerned firstly the use of different elicitation types by teachers to stimulate pupils into giving responses and secondly teachers' recourse to code-switching from English to Kiswahili and vice-versa. It was also found in this study that although the use of English as the medium of instruction in the classrooms which were observed resulted in certain communication problems, some of these problems need not have arisen if teachers had been more careful in their use of language. The consideration of this finding, after taking into account the role of different elicitation types and code-switching as interpretable from data samples had certain implications which are specified in the study for teaching in Kenyan primary schools. The corpus for the study consisted of audio-recordings of English, Science and Number-Work lessons which were later transcribed. Relevant data samples were subsequently extracted from transcripts for analysis. Many of the samples have examples of cases of communication breakdowns, but they also illustrate how teachers maintained interaction with pupils who had yet to acquire an operational mastery of English. This study thus differs from most studies on classroom interaction because of its basic concern with the examination of the resources available to teachers for overcoming the problem areas of classroom communication.

Organisational change and safety culture: the impact of communication

This research examines and explains the links between safety culture and communication. Safety culture is a concept that in recent years has gained prominence but there has been little applied research conducted to investigate the meaning of the concept in 'real life' settings. This research focused on a Train Operating Company undergoing change in a move towards privatisation. These changes were evident in the management of safety, the organisation of the industry and internally in their management. The Train Operating Company's management took steps to improve their safety culture and communications through the development of a cascade communication structure. The research framework employed a qualitative methodology in order to investigate the effect of the new system on safety culture. Findings of the research were that communications in the organisation failed to be effective for a number of reasons, including both cultural and logistical problems. The cultural problems related to a lack of trust in the organisation by the management and the workforce, the perception of communications as management propaganda, and asyntonic communications between those involved, whilst logistical problems related to the inherent difficulties of communicating over a geographically distributed network. An organisational learning framework was used to explain the results. It is postulated that one of the principal reasons why change, either to the safety culture or to communications, did not occur was because of the organisation's inability to learn. The research has also shown the crucial importance of trust between the members of the organisation, as this was one of the fundamental reasons why the safety culture did not change, and why safety management systems were not fully implemented. This is consistent with the notion of mutual trust in the HSC (1993) definition of safety culture. This research has highlighted its relevance to safety culture and its importance for organisational change.

Deconstruction-analysis-explanation:contextualization in professional digital discourse

It is already a truism that emerging communication technologies have changed the landscape of communication in every aspect of our lives, but this is specifically true for how we communicate at work. Advances in communication technologies have enabled a wide range of digital communication modes to be utilized for both internal and external business communication; including audio and visual communication and voice-over protocols, as well as text-based channels, such as email, forums, instant messaging and social media. In spite of the wide range of available audio-visual channels, and despite the ever-increasing popularity of email, real-time text-based communication technologies (instant messaging or IM) are also on the rise (see Mak, 2014; Pazos et al., 2013; Radicati & Levenstein, 2013; and Markman in this volume). The prominence of IM is evident in the rise of this mode of communication, not only as a tool for internal business communication, but as a front-stage channel, particularly for customer service encounters or professional-client conversations (Makarem et al., 2009; Pearce et al., 2013; L. Zhang et al., 2011).

Identifying criteria for the assessment of pharmacy students' communication skills with patients

Objectives To identify criteria by which patients can assess the communication skills of pharmacy students. Method Potential assessment criteria were generated from 2 main sources: a literature review and a focus group discussion. A modified two-round Delphi survey was subsequently conducted with 35 professionals who were actively involved in teaching and assessing communication skills of pharmacy students to determine the importance and reliability of each criterion. Results Consensus ratings identified 7 criteria that were important measures of pharmacy students' communication skills and could be reliably assessed by patients. Conclusions A modified two-round Delphi consultation survey successfully identified criteria that can be used by patients to assess the communication skills of pharmacy undergraduates. Future work will examine the feasibility of using patients as assessors of communication skills of pharmacy students, preregistration pharmacists, and qualified pharmacists.

Determinants of entry mode choice of MNCs in emerging markets:evidence from South Africa and Egypt

It is now stylized that the importance of foreign direct investment for developing countries and emerging markets arises from the impact of the presence of multinational corporations (MNCs) in the host country on the productivity of local firms, by way of technology diffusion and competition. There is also general agreement that the extent of technology transfer by an MNC to a developing country affiliate depends on the extent of its control on the local affiliate and that, in turn, the extent of this control depends on the mode of entry of the MNC into the host country. However, the existing literature is based on the experience of developed countries and as such does not contribute to the literature on development economics. This article addresses this lacuna using unique firm-level data from South Africa and Egypt. Our results indicate that the determinants of entry mode choice not only differ between developed and developing countries, but also among developing countries. They also bring into question the role of MNCs in fostering productivity growth in developing countries.

Stabilization of a passively mode-locked laser by continuous wave optical injection

We investigate numerically and experimentally the properties of a passively mode locked quantum dot semiconductor laser under the influence of cw optical injection. We demonstrate that the waveform instability at high pumping for these devices can be overcome when one mode of the device is locked to the injected master laser and additionally show spectral narrowing and tunability. Experimental and numerical analyses demonstrate that the stable locking boundaries are similar to these obtained for optical injection in CW lasers. © 2010 American Institute of Physics.

Analyze impact of context information on rate adaptation algorithm in vehicular communication

Nowadays, road safety and traffic congestion are major concerns worldwide. This is why research on vehicular communication is very vital. In static scenarios vehicles behave typically like in an office network where nodes transmit without moving and with no defined position. This paper analyses the impact of context information on existing popular rate adaptation algorithms. Our simulation was done in MATLAB by observing the impact of context information on these algorithms. Simulation was performed for both static and mobile cases.Our simulations are based on IEEE 802.11p wireless standard. For static scenarios vehicles do not move and without defined positions, while for the mobile case, vehicles are mobile with uniformly selected speed and randomized positions. Network performance are analysed using context information. Our results show that in mobility when context information is used, the system performance can be improved for all three rate adaptation algorithms. That can be explained by that with range checking, when many vehicles are out of communication range, less vehicles contend for network resources, thereby increasing the network performances. © 2013 IEEE.