Development of immunoassays for the detection of the fungicide penconazole and its urinary metabolite.

Monoclonal antibodies were raised to haptens containing moieties common to both the triazole fungicide penconazole and its proposed primary urinary metabolite (4-(2,4-dichlorophenyl)-5-(H-1,2,4-triazol-1-yl)pentoic acid). The monoclonal antibody 2E4 was used to develop competitive ELISA assays where binding of antibody to immobilized haptens conjugated to BSA competed with penconazole or its metabolite in solution. At pH 4.0 and pH 8.0, penconazole was detected with an IC50 of 1.0-1.2 microg/L respectively and at pH 4 penconazole metabolite was detected with an IC50 of 0.9 microg/L. These assays were specific for penconazole and/or its metabolite compared to other triazole fungicides. The immunoassay conditions optimal for penconazole metabolite (pH 4.0) were used and applied to the analysis of spiked human urine, and following sample extraction using a C18 SPE column, could detect 0.5 microg/L metabolite. This is the first report of an immunoassay to the urinary metabolite of penconazole, an assay with application in the monitoring of occupational and non-occupational exposure to this commonly used pesticide.

Kinase array design, back to front: biaryl amides.

New kinase inhibitors can be found by synthesis of targeted arrays of compounds designed using system-based knowledge as well as through screening focused or diverse compounds. Most array strategies aim to add functionality to a fragment that binds in the purine subpocket of the ATP-site. Here, an alternative pharmacophore-guided array approach is described which set out to discover novel purine subpocket-binding groups. Results are shown for p38alpha and cFMS kinase, for which multiple distinct series with nanomolar potency were discovered. Some of the compounds showed potency in cell-based assays and good pharmacokinetic properties.

Biphenyl amide p38 kinase inhibitors 3: Improvement of cellular and in vivo activity.

The biphenyl amides (BPAs) are a novel series of p38alpha MAP kinase inhibitor. The optimisation of the series to give compounds that are potent in an in vivo disease model is discussed. SAR is presented and rationalised with reference to the crystallographic binding mode.

Managing MRSA in hospital and in the community.

Shila Patel explains the different management protocols for controlling an outbreak of MRSA in hospital and the community.

Principles of appropriate use of disposable gloves.

Shila Patel explains the principles of glove use, highlighting the procedures for which they are indicated and advising on appropriate storage.

Recombinant anti-EspA antibodies block Escherichia coli O157:H7-induced attaching and effacing lesions in vitro.

Intimin and EspA proteins are virulence factors expressed by attaching and effacing Escherichia coli (AEEC) such as enteropathogenic and enterohaemorrhagic E. coli. The EspA protein makes up a filament structure forming part of the type III secretion system (TTSS) that delivers effector proteins to the host epithelial cell. Bacterial surface displayed intimin interacts with translocated intimin receptor in the host cell membrane leading to intimate attachment of the bacterium and subsequent attaching and effacing lesions. Here, we have assessed the use of recombinant monoclonal antibodies against E. coli O157:H7 EspA and intimin for the disruption of AEEC interaction with the host cell. Anti-gamma intimin antibodies did not reduce either adhesion of E. coli O157:H7 to host cell mono-layers or subsequent host cell actin rearrangement. Anti-EspA antibodies similarly had no effect on bacterial adhesion however they had a marked effect upon E. coli O157:H7-induced host cell actin rearrangement, where both monoclonal and polyclonal antibodies completely blocked cytoskeletal changes within the host cell. Furthermore, these anti-EspA antibodies were shown to reduce actin rearrangement induced by some but not all other AEEC serotypes tested. Both polyclonal and monoclonal antibodies could be used to label E. coli O157 EspA filaments and these immunoreagents did not inhibit the formation of such filaments. This is the first report of monoclonal antibodies to EspA capable of disrupting the TTSS function of E. coli O157:H7.

Minimising cross-infection risks associated with beds and mattresses.

The spread of infection can be associated with inanimate objects in the environment, referred to as fomites. These include beds, mattresses, curtains, bedpans and sphygmomanometers, all of which may be contaminated with micro-organisms (Wilson, 2001).

The efficacy of alcohol-based hand disinfectant products.

Over the last few years alcohol-based hand disinfectants have become widely available within health care, providing an alternative means of achieving good hand decontamination. In the hospital setting their advantage over soap and water is that they can be applied in transit to the next patient or task and therefore may help improve compliance with hand decontamination. Within the community setting they provide a suitable alternative to handwashing, particularly where there may be inadequate handwashing facilities. This article considers some issues around their use, namely indications for use, efficacy, and potential for skin damage.

The impact of environmental cleanliness on infection rates.

Much has been said in recent years about the lack of environmental cleanliness in hospitals. The House of Lords Select Committee (1998) talked about falling standards in hospital cleaning and, more recently, the government acknowledged in The NHS Plan (Department of Health, 2000) that hospitals were unacceptably dirty. Such comments may affect people's perceptions of hospitals and, rather than regarding them as safe environments that promote healing, they may be viewed as dangerous places that might cause vulnerable patients greater harm. While it is known that about 10 per cent of hospital inpatients have a nosocomial infection at any one time (Emmerson et al, 1996), there is uncertainty about whether there is a sound evidence base to support the theory that dirty hospitals cause infection.

Examining the literature on using tap water in wound cleansing.

Practitioners working in the community setting and whose caseload predominantly consists of treating chronic wounds often use tap water as a wound cleanser with the rationale that chronic wounds are already colonised with bacteria. However, there is some controversy, as discussed within this review, of the suitability of tap water as a cleansing agent for acute wounds.