Contributions of two UDP-glucose dehydrogenases to viability and polymyxin B resistance of Burkholderia cenocepacia

Burkholderia cenocepacia is highly resistant to antimicrobial peptides and we hypothesized that the conversion of UDP-glucose to UDP-glucuronic acid, a reaction catalysed by the enzyme UDP-glucose dehydrogenase (Ugd) would be important for this resistance. The genome of B. cenocepacia contains three predicted ugd genes: ugd(BCAL2946), ugd(BCAM0855) and ugd(BCAM2034), all of which were individually inactivated. Only inactivation of ugd(BCAL2946) resulted in increased sensitivity to polymyxin B and this sensitivity could be overcome when either ugd(BCAL2946) or ugd(BCAM0855) but not ugd(BCAM2034) was expressed from plasmids. The growth of a conditional ugd(BCAL2946) mutant, created in the Deltaugd(BCAM0855) background, was significantly impaired under non-permissive conditions. Growth could be rescued by either ugd(BCAL2946) or ugd(BCAM0855) expressed in trans, but not by ugd(BCAM2034). Biochemical analysis of the purified, recombinant forms of Ugd(BCAL2946) and Ugd(BCAM0855) revealed that they are soluble homodimers with similar in vitro Ugd activity and comparable kinetic constants for their substrates UDP-glucose and NAD(+). Purified Ugd(BCAM2034) showed no in vitro Ugd activity. Real-time PCR analysis showed that the expression of ugd(BCAL2946) was 5.4- and 135-fold greater than that of ugd(BCAM0855) and ugd(BCAM2034), respectively. Together, these data indicate that the combined activity of Ugd(BCAL2946) and Ugd(BCAM0855) is essential for the survival of B. cenocepacia but only the most highly expressed ugd gene, ugd(BCAL2946), is required for polymyxin B resistance.

A putative gene cluster for aminoarabinose biosynthesis is essential for Burkholderia cenocepacia viability

Using a conditional mutagenesis strategy we demonstrate here that a gene cluster encoding putative aminoarabinose (Ara4N) biosynthesis enzymes is essential for the viability of Burkholderia cenocepacia. Loss of viability is associated with dramatic changes in bacterial cell morphology and ultrastructure, increased permeability to propidium iodide, and sensitivity to sodium dodecyl sulfate, suggesting a general cell envelope defect caused by the lack of Ara4N.

Chymotrypsin-like serine proteinases are involved in the maintenance of cell viability

An increasing number of studies have implicated serine proteinases in the development of apoptosis. In this study, we assessed the ability of a set of highly specific irreversible inhibitors (activity probes), incorporating an a-amino alkane diphenyl phosphonate moiety, to modulate cell death. In an initial assessment of the cellular toxicity of these activity probes, we discovered that one example, N-a-tetramethylrhodamine phenylalanine diphenylphosphonate {TMR-Phe^P(OPh)₂} caused a concentration-dependent decrease in the viability of HeLa and U251 mg cells. This reduced cell viability was associated with a time-dependent increase in caspase-3 activity, PARP cleavage and phosphatidylserine translocation, establishing apoptosis as the mechanism of cell death. SDS-PAGE analysis of cell lysates prepared from the HeLa cells treated with TMR-Phe^P(OPh)₂, revealed the presence of a fluorescent band of molecular weight 58 kDa. Given that we have previously reported on the use of this type of activity probe to reveal active proteolytic species, we believe that we have identified a chymotrypsin-like serine proteinase activity integral to the maintenance of cell viability.

More equitable systems for allocating general practice deprivation payments:financial consequences

The allocation of general practitioner (GP) deprivation payments has been a controversial topic since they were first proposed. It has recently been suggested that the current system could be made more equitable if the payments were allocated at enumeration districts and if there was a more graded relationship between Jarman score and funding. However, the implications of these changes on the distribution of deprivation payments have not been worked out.

Interlocking directorates and conflicts of interest: The Rotterdamsche Bankvereeniging, Müller & Co. and the Dutch financial crisis of the 1920s

How can interlocking directorates cause financial instability for universal banks? A detailed history of the Rotterdamsche Bankvereeninging in the 1920s answers this question in a case study. This large commercial bank adopted a new German-style universal banking business model from the early 1910s, sharing directors with the firms it financed as a means of controlling its interests. Then, in 1924, it required assistance from the Dutch state in order to survive a bank run brought on by public concerns over its close ties with Müller & Co., a trading conglomerate that suffered badly in the economic downturn of the early 1920s. Using a new narrative history combined with an interpretive model, this article shows how the interlocking directorates between the bank and this major client, and in particular the direction of influence of these interlocks, resulted in a conflict of interest that could not be easily overcome.

The rebirth of dependence– offering an alternative understanding of the financial crisis

Purpose – This article aims to contribute to the re-evaluation of the global market system using a Marxist inspired theory of development, dependency.

Design/methodology/approach – This article draws on dependency theory as an alternative means of understanding global relationships. Building on existing literature, it modifies dependency to encapsulate technological developments and trends in the global market.

Findings – Re-evaluating the global market and the relationships that underpin it, through an alternative theory, highlights the fragility of markets and associated relationships. Increasingly, nation states are becoming irrelevant. This presents a problem as the main actors in the global market today are “above” inter-state relations, yet the organs that regulate their behaviour still are grounded in inter-state rhetoric. The relationship between development and underdevelopment remains.

Research limitations/implications – The financial crisis has propagated a wealth of interest in the relationships between states, between multi-national corporations (MNCs) and between MNCs and state. Using this broad theory of modified dependency, it can be applied to a range of different relationships. In the wake of financial crisis, there is the opportunity to raise awareness of these ingrained issues and initiate discussions at national, regional and international levels to alleviate some of the conditions of dependence.

Practical implications – Regardless of the work of national governments and NGOs to instigate development in lesser-developed regions through policy and regulations, unless there is a conscientious commitment from MNCs operating in that region to contribute to development, the result will be the development of underdevelopment and the underdevelopment of development. CSR can help alleviate the conditions of the dependence on capital generated by MNCs, but this is not a solution to an ingrained problem, capitalism.

Originality/value – This article introduces a modified theory of dependency for the first time. It applies the theory to the financial crisis and to the continent of Africa. It considers the role that CSR can play in alleviating the conditions of dependence.