The Host Defence Peptide LL-37 is Susceptible to Proteolytic Degradation by Wound Fluid Isolated from Foot Ulcers of Diabetic Patients

The pleiotropic effects of host defence peptides (HDPs), including the ability to kill microorganisms, enhance re-epithelialisation and increase angiogenesis, indicates a role for these important peptides as potential therapeutic agents in the treatment of chronic, non-healing wounds. However, the maintenance of peptide integrity, through resistance to degradation by the array of proteinases present at the wound site, is a prerequisite for clinical success. In this study we explored the degradation of exogenous LL-37, one such HDP, by wound fluid from diabetic foot ulcers to determine its susceptibility to proteolytic degradation. Our results suggest that LL-37 is unstable in the diabetic foot ulcer microenvironment. Following overnight treatment with wound fluid, LL-37 was completely degraded. Analysis of cleavage sites suggested potential involvement of both host- and bacterial-derived proteinases. The degradation products were shown to retain some antibacterial activity against Pseudomonas aeruginosa but were inactive against Staphylococcus aureus. In conclusion, our data suggest that stabilising selected peptide bonds within the sequence of LL-37 would represent an avenue for future research prior to clinical studies to address its potential as an exogenously-applied therapeutic in diabetic wounds. 

Attitudes of Irish dental graduates to vocational training

INTRODUCTION: Vocational training (VT) is a mandatory 12 month period for UK dental graduates after graduation. Graduates of Irish Dental Schools are eligible to enter the general dental service in Ireland or obtain an NHS performers list number in the UK immediately after qualification. Reports would suggest that some graduates of Irish Dental Schools are choosing to take part in VT in the UK and find the experience beneficial. This study aimed to record the uptake of VT amongst recent graduates from University College Cork and to document their experiences. It was designed to compare the attitudes and experiences of graduates of Irish Dental Schools who undertook VT compared with those who entered the general dental service.

METHOD: A self-completion questionnaire was distributed by e-mail to dental graduates from University College Cork who had graduated 2001-2007. Responses were returned by e-mail or post.

RESULTS: The response rate was 68.9%. There has been an increase in the numbers of graduates taking part in VT each year since 2004. 92.5% of Vocational Dental Practitioners (VDPs) found their experience beneficial as they received a guaranteed source of income, had a supportive peer network and worked in a positive learning environment. However, some felt that they earned a lower income than their associate colleagues, others found the pace of practice slow and that the duration of the training period was excessive. Eighty-five per cent of VDPs would choose the same position again after graduation as compared with 61.8% of associates (P < 0.001). Ninety per cent of VDPs would advise current undergraduates to take part in VT as compared with 51% of associates (P < 0.001). A larger proportion of VDPs had taken part in postgraduate studies but there was no significant difference between the two groups.

CONCLUSIONS: Larger proportions of recent graduates are undertaking vocational training.--The majority of VDPs and associates find their initial employment position beneficial.--VDPs benefit from a guaranteed source of income, a supportive peer network and a positive learning environment.--Some associates suffered from a lack of support, feeling isolated and overwhelmed with patients.--The majority of previous VDPs and associates would recommend VT to current undergraduates.--Almost 40% of associates would now choose to take part in VT if given the opportunity.

Mechanisms of reduced susceptibility and genotypic prediction of antibiotic resistance in Prevotella isolated from cystic fibrosis (CF) and non-CF patients

OBJECTIVES: To investigate mechanisms of reduced susceptibility to commonly used antibiotics in Prevotella cultured from patients with cystic fibrosis (CF), patients with invasive infection and healthy control subjects and to determine whether genotype can be used to predict phenotypic resistance.

METHODS: The susceptibility of 157 Prevotella isolates to seven antibiotics was compared, with detection of resistance genes (cfxA-type gene, ermF and tetQ), mutations within the CfxA-type β-lactamase and expression of efflux pumps.

RESULTS: Prevotella isolates positive for a cfxA-type gene had higher MICs of amoxicillin and ceftazidime compared with isolates negative for this gene (P < 0.001). A mutation within the CfxA-type β-lactamase (Y239D) was associated with ceftazidime resistance (P = 0.011). The UK CF isolates were 5.3-fold, 2.7-fold and 5.7-fold more likely to harbour ermF compared with the US CF, UK invasive and UK healthy control isolates, respectively. Higher concentrations of azithromycin (P < 0.001) and clindamycin (P < 0.001) were also required to inhibit the growth of the ermF-positive isolates compared with ermF-negative isolates. Furthermore, tetQ-positive Prevotella isolates had higher MICs of tetracycline (P = 0.001) and doxycycline (P < 0.001) compared with tetQ-negative isolates. Prevotella spp. were also shown, for the first time, to express resistance nodulation division (RND)-type efflux pumps.

CONCLUSIONS: This study has demonstrated that Prevotella isolated from various sources harbour a common pool of resistance genes and possess RND-type efflux pumps, which may contribute to tetracycline resistance. The findings indicate that antibiotic resistance is common in Prevotella spp., but the genotypic traits investigated do not reflect phenotypic antibiotic resistance in every instance.