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Chronic Pseudomonas aeruginosa infection in cystic fibrosis (CF) leads to a damaging host inflammatory response. There are an increasing number of reports of P. aeruginosa cross-infection at CF centres. The clinical significance of acquisition of a transmissible strain for patients who already harbour P. aeruginosa is unclear. In this study, levels of inflammatory markers in clinically stable adult CF patients who harbour transmissible and sporadic strains of P. aeruginosa have been compared. Patients with CF and chronic P. aeruginosa infection were grouped into those who harbour a transmissible P. aeruginosa and those who harbour their own sporadic strains. Total white cell and differential counts, sputum neutrophil elastase (NE), interleukin (IL)-8, tumour necrosis factor (TNF)-alpha, plasma IL-6 and NE/alpha1-antitrypsin complexes, serum C-reactive protein, and urine TNF receptor 1 were all measured in clinically stable patients 4-6 weeks following completion of intravenous antibiotic therapy. The two groups (both n=20) were well matched for per cent predicted forced expiratory volume in one second, per cent predicted forced vital capacity and body mass index. There were no significant differences in levels of white cell counts or inflammatory markers between the two groups. At times of clinical stability, cystic fibrosis patients infected with transmissible Pseudomonas aeruginosa do not have a heightened inflammatory response above that of those harbouring sporadic strains.

B ARRY F., G oRG H. and M CDOWELL A. (2003) Outward FDI and the investment development path of a late-industrializing economy: evidence from Ireland, Reg. Studies 37 , 341-349. The Investment Development Path (IDP) hypothesis holds that a country's net outward direct investment position is systematically related to its level of economic development. Ireland is an interesting test case because of the importance of inward foreign direct investment (FDI) over the last three decades and the recent increase in outward FDI by Irish-owned multinationals. We find empirical support for the IDP concept for the Irish case. Our sectoral analysis suggests that Ireland's outward FDI flows are disproportionately horizontal and oriented towards non-internationally-tradable sectors. Also, the firm-specific assets of Irish multinationals lie neither in R&D nor in the type of product differentiation associated with high advertising expenditures. B ARRY F., G oRG H. et M CDOWELL A. (2003) L'investissement direct a l'etranger...

‘Belief,’ along with ‘believe, beliefs,’ and ‘belief systems,’ has served as a kind of ‘odd job’ word for anthropologists: a word commonly used for the analysis of a society's culture, religion, or ideas about the world, but seldom defined or explicitly theorized. Belief is often included in omnibus definitions of culture, such as Tylor's classic “Culture, or civilization, … is that complex whole, which includes knowledge, belief, art, law, morals, custom, and any other capabilities and habits acquired by man as a member of society” (1871, from Kroeber and Kluckhohn, 1952, p. 43) or Sapir's “culture, that is, … the socially inherited assemblage of practices and beliefs that determines the texture of our lives …” (1921, Kroeber and Kluckhohn, 1952, p. 47). The term often refers explicitly to religion, as in Tylor's minimum definition of religion as ‘the belief in Spiritual Beings’ (1873), but is used equally for claims people make about the empirical world or their social institutions.

From a medical microbiology perspective, the species Propionibacterium acnes, Propionibacterium avidum, Propionibacterium granulosum, and Propionibacterium propionicum, are the most relevant within the genus Propionibacterium. Of these, Propionibacterium acnes has the greatest pathogenic potential and is associated with a diverse range of infections and clinical conditions, ranging from inflammatory acne to prosthetic joint infections and prostate cancer. In this chapter, we provide an overview of the genus Propionibacterium, and describe the role played by the ‘cutaneous’ propionibacteria, especially P. acnes, in human disease. We also cover virulence factors, mechanisms of antibiotic resistance, insights from whole-genome sequencing and immune responses. The potential therapeutic effects of propionibacteria when used as adjuvants/vaccines are also considered, along with molecular-based approaches for differentiation and characterization of propionibacteria associated with human infection.

Musculoskeletal diseases such as rheumatoid arthritis are complex multifactorial disorders which are chronic in nature and debilitating for patients. A number of drug families are available to clinicians to manage these disorders but few tests exist to target these to the most responsive patients. Therefore drug failure and switching to drugs with alternate modes of action is common place. In parallel, a limited number of laboratory tests are available which measure biological indicators or ‘biomarkers’ of disease activity, autoimmune status, or joint damage. There is a growing awareness that assimilating the fields of drug selection and diagnostic tests into ‘companion diagnostics’ could greatly advance disease management and improve outcomes for patients. This review aims to highlight: (i) the current applications of biomarkers in rheumatology with particular focus on companion diagnostics, (ii) developments in the fields of proteomics, genomics, the microbiome, imaging and bioinformatics, (iii) how integration of these technologies into clinical practice could support therapeutic decisions.

This article reexamines medical anthropology theories of symptom, illness, and disease to consider unregulated medical care in India. It builds on clinical observations, an inventory of the pharmaceuticals used by men who call themselves "Bengali doctors," and their patients to understand medical care in a context that privileges symptom not disease. It draws on Derrida's use of pharmakon to outline the complexities of care and embodiment and helps locate local and medical anthropology theories of symptom and pharmaceuticals within theories of the experiential body. It asks two key questions: What is medical care without disease and what are its implications on a local biology in which disease-based biomedicine is modified? Searching for a tentative answer, it works to bring medical anthropology's interest in symptom back to the body without losing symptoms' connection to political economies, individual experience, and localized biomedicine.

Propionibacterium acnes and coagulase-negative staphylococci (CoNS) are opportunistic pathogens implicated in prosthetic joint and fracture fixation device-related infections. The purpose of this study was to determine whether P. acnes and the CoNS species Staphylococcus lugdunensis , isolated from an “aseptically failed” prosthetic hip joint and a united intramedullary nail-fixed tibial fracture, respectively, could cause osteomyelitis in an established implant-related osteomyelitis model in rabbits in the absence of wear debris from the implant material. The histological features of P. acnes infection in the in vivo rabbit model were consistent with localized pyogenic osteomyelitis, and a biofilm was present on all explanted intramedullary (IM) nails. The animals displayed no outward signs of infection, such as swelling, lameness, weight loss, or elevated white blood cell count. In contrast, infection with S. lugdunensis resulted in histological features consistent with both pyogenic osteomyelitis and septic arthritis, and all S. lugdunensis -infected animals displayed weight loss and an elevated white blood cell count despite biofilm detection in only two out of six rabbits. The differences in the histological and bacteriological profiles of the two species in this rabbit model of infection are reflective of their different clinical presentations: low-grade infection in the case of P. acnes and acute infection for S. lugdunensis . These results are especially important in light of the growing recognition of chronic P. acnes biofilm infections in prosthetic joint failure and nonunion of fracture fixations, which may be currently reported as “aseptic” failure.

A series of electrophilic glutamine analogues based on 6-diazo-5-oxo-norleucine has been prepared, using novel synthetic routes, and evaluated as inhibitors of Escherichia coli glucosamine synthetase. The gamma-dimethylsulphonium salt analogue of glutamine was found to be one of the most potent inactivators of this enzyme yet reported, with an apparent second order rate constant (k2/Ki) of 3.5 x 10(5) M(-1) min(-1).

While the association of the Gram-positive anaerobic bacterium Propionibacterium acnes with the inflammatory skin condition acne vulgaris has been known for over a century, its potential role in other human infections and clinical conditions has undoubtedly been underestimated. This is likely to reflect inadequate anaerobic processing methods and culture of clinical specimens (>7d incubation), combined with the traditionally held view that P. acnes in a biological sample simply reflects contamination from the resident skin microflora or is clinically irrelevant due to a low level of virulence [1]. On such basis, it is likely that an unknown number of P. acnes infections go undiagnosed and unrecognised. In addition, subtle clinical signs and symptoms that are often associated with P. acnes infections may further lead to the conclusion that an infection is absent, particularly in relation to prosthetic joints and other indwelling medical devices [2]. Despite the historical failure to recognise the pathogenic potential of P. acnes, there is now a growing awareness, as evidenced in the scientific literature, that the bacterium is an important cause of opportunistic human infection and should not be readily dismissed as a contaminant/passive presence in clinical samples [2–6]. Within the last decade, advances in our understanding of the population genetic structure of P. acnes combined with whole genome sequence (WGS) data now available for a large number of strains, mostly derived from the Human Microbiome Project, have provided a platform for hypothesis-driven research into the exact role of P. acnes in human health and disease and the possibility of developing vaccines and other novel therapeutic treatments [7–11]. In particular, the availability of WGS information has enabled both genomic and transcriptomic differences between lineages from phylogroups associated mainly with health or disease to be examined, thus identifying potential genetic determinants and mechanisms of virulence [11, 12]. Despite its ability to behave as an opportunistic pathogen, it must not be forgotten that P. acnes is part of the normal human microbiota and, consequently, also plays a role in human health via occupation of niches that could be colonized by other, more pathogenic, microorganisms. Furthermore, the powerful immunomodulatory properties of P. acnes have long been investigated for their capacity to stimulate protective host responses against various human cancers, and more generally against Th-2-mediated diseases. In light of the growing interest and recognition of the role of P. acnes in human health and disease, this special issue aims to provide a description of recent developments and advances in different areas of P. acnes research via original research and review articles, and thus provide a framework to discuss and identify key issues that need to be addressed in future studies. Developments in our understanding of P. acnes bacteriophages are reviewed in this special issue by H. Bruggemann and R. Lood. They describe both historical and recent developments in the field and highlight how genomic analyses have provided insights into P. acnes phage biology, as well as the identification of phage sequence signatures in clustered regularly interspaced short palindromic repeats (CRISPRs) in P. acnes. They also discuss bacteriophages as a therapeutic strategy to combat P. acnes-associated diseases, although there are currently formidable regulatory barriers to this approach. In a research paper, G. Kasimatis et al. describe a comparative genomics study on the acne-derived type IA1 strain, HL096PA1, which they found harbours the first described linear plasmid in P. acnes, pIMPLE-HL096PA1. Homologues of pIMPLE-HL096PA1 appear to be widely distributed in ribotype 4 and 5 strains of P. acnes which are tetracycline resistant and associated with acne [11]. These plasmids carry a tight adhesion locus (Tad) that may enhance binding to host cell surfaces. Identification of this novel plasmid also provides a potentially new opportunity for genetic manipulation of P. acnes and possibly targeted therapy against specific disease-associated strains. HL096PA1 also contains fivefold more pseudogenes than the type IB strain KPA171202 to which it was compared, as well as island-like regions unique to the strain; type IB strains have rarely been isolated from acne lesions. Collectively, this paper provides additional data on P. acnes virulence properties and host adaptation mechanisms that may be relevant to our understanding of acne pathogenesis. Still on the theme of acne, E. A. Eady et al. in a second review paper hypothesise that the availability of water and possibly one or more water soluble micronutrients limits microbial growth within the follicular microenvironment of the skin and that reduction of water activity with biocompatible solutes of very high solubility may represent a novel and safe therapeutic approach in acne management. In an era of growing antibiotic resistance and the continuing widespread use of oral and topical antibiotics for the first line treatment of acne, the development of alternative therapeutic approaches for this extremely common skin condition is highly desirable. In this special issue, we also have a research paper by N. Fischer et al. which describes a study into the intracellular fate of P. acnes in macrophages. This is an important study as the ability to persist in macrophages and other cell types as an intracellular pathogen may be important in the context of P. acnes-related diseases, particularly acne, sarcoidosis, and prostate cancer, to which the bacterium has been associated. This study confirmed the ability of P. acnes to survive phagocytosis in vitro, although no intracellular replication or escape from THP-1 host cells were observed. There was also no evidence of the P. acnes-containing phagosome fusing with lysosomes. The association of P. acnes with sarcoidosis, a systemic granulomatous disease, is extensively reviewed in a paper by Y. Eishi. While sarcoidosis is a disease of unknown etiology, infectious granulomas are commonly caused by intracellular pathogens and P. acnes is currently the only microorganism isolated from sarcoid lesions by culture. The review proposes mechanisms of granuloma formation in response to P. acnes in subjects with sarcoidosis and introduces a new concept of endogenous infection caused by hypersensitivity to indigenous bacteria. Interestingly, acne is also a delayed type hypersensitivity reaction, and if the condition is left untreated granuloma formation can occur [13]; this is rarely seen today due to clinical intervention. In a research paper by I. Dekio et al., the intracellular proteomes of different phylogroups were investigated and variations in protein expression profiles between strains grown under different oxygen tensions were examined. A good correlation was observed between genomic and proteomic profiles for the major genetic divisions of P. acnes. Differences were also noted in the intracellular proteome of strains grown under anaerobic/microaerophilic and aerobic culture conditions, which could be relevant in the context of P. acnes transferring from the skin surface to hypoxic/anoxic environments associated with deep seated systemic infections. The role of P. acnes as a cause of infections related to indwelling medical devices is now increasingly recognised, especially in relation to prosthetic shoulder implants [2, 14]. A study from the Mayo Clinic, described by M. J. Karau et al., highlights how application of a vortexing-sonication technique for the detection of bacterial biofilm identifies Propionibacterium species, likely P. acnes, as common colonizers of breast tissue expanders. The presence of these colonised expanders in clinically uninfected subjects may present an increased risk factor for future infection or capsular contracture. The role of P. acnes in a wide range of implant-associated infections, along with its clinical management, is further discussed in a review by M. Portillo et al. They also describe how sonication of explanted prosthetic material improves the diagnosis of such infections and suggest that molecular methods may further increase the sensitivity of P. acnes detection in the resulting sonicate. In a paper by J. Rollason et al., P. acnes isolates from surgically excised lumbar disc herniations are characterised for phylogroup and antimicrobial susceptibility. Previous studies have suggested a role for P. acnes in the pathophysiology of disc degeneration and herniation, and antibiotic treatment does appear to have a positive effect upon patients with chronic back conditions [15–17]. In this study, type II and III strains were the predominant phylotypes isolated. Since strains from these genetic divisions are infrequently recovered from the skin, this study suggests that the recovery of P. acnes from excised disc material should not be readily dismissed as skin contamination in all cases. Furthermore, isolates from excised discs were susceptible to a range of antimicrobials commonly used to treat P. acnes-related infections. In summary, this special issue covers a range of diverse topics related to P. acnes and disease, thus highlighting the widespread opportunistic behaviour of the bacterium in relation to infection. We hope the papers published will serve to further highlight the pathogenic potential of P. acnes, as well as stimulating further research into the role of the bacterium in human health and disease. In particular, future studies aimed at providing firm evidence of a role in chronic diseases, such as prostate cancer and disc disease, may provide exciting new opportunities for novel patient treatment and management strategies.

Landscape planners and designers are interested in replicating natural landscape patterns to reclaim degraded landscapes to blend with existing conditions. One approach that shows promise is the use of fractal geometry to create natural landscape patterns. While the measurement of the actual fractal dimension of an object is difficult, the box-counting method (developed at Agrocampus Ouest, Angers, France) approximates the fractal dimension of an object. This process is illustrated by measuring and replicating a stand of trees in the Upper Peninsula of Michigan and applying the method for a planting plan on a Northern Michigan surface mine. The estimated fractal dimensions for the tree species are calculated: 0.329 for Tsuga canadensis Carriere, 0.674 for Thuja occidentalis L., 0.607 for Acer rubrum L, 0.345 for Acer saccharum Marshall, 0.442 for Pinus strobus L., and 0.359 for Picea glauca (Moench) Voss. and were applied in the design of a revegetation plan.

Utilising strategically positioned bed-mounted accelerometers, the Passive Sleep Actigraphy platform aims to deliver a non-contact method for identifying periods of wakefulness during night-time sleep. One of the key problems in developing data driven approaches for automatic sleep monitoring is managing the inherent sleep/wake class imbalance. In the current study, actigraphy data from three participants over a period of 30 days was collected. Upon examination, it was found that only 10% contained wake data. Consequently, this resulted in classifier overfitting to the majority class (sleep), thereby impeding the ability of the Passive Sleep Actigraphy platform to correctly identify periods of wakefulness during sleep; a key measure in the identification of sleep problems. Utilising Spread Subsample and Synthetic Minority Oversampling Techniques, this paper demonstrates a potential solution to this issue, reporting improvements of up to 28% in wake detection when compared to baseline data while maintaining an overall classifier accuracy of 90%.

Over 30 years ago Stott and Davis1 described four types of activities in the general practice consultation, in which opportunistic health promotion has a key place (and is still the backbone of the QOF). Five years later, Pendleton2 described a time-sequenced, task-based model of the consultation in which emphasised the importance of exploring patients’ ideas, concerns, and expectations. Neighbour, in 1987,3 saw the process of consultation as climbing a mountain, describing five stages in the ascent. He identified the need for safety-netting, ensuring that future risks to the patient and other diagnoses are considered, and housekeeping, where, after the consultation, the doctor deals with their own feelings. Middleton’s4 process model is not time sequenced, but describes something of what Neighbour called connecting with the patient, and is also a helpful way to look at Pendleton’s ideas, concerns, and expectations in more depth. Middleton acknowledges that both patients and doctors have agendas and that by clarifying both, at least in the mind of the doctor, it is possible to gain a better understanding of the consultation. Silverman et al 5 produced a list of 54 skills to deploy in the consultation, with a consultation framework to put them in. Along with the teaching model developed to train doctors, these skills have dominated thinking on the consultation for 15 years. Chaos theory6 has taught us something about how to deal with complex consultations when the number of uncertainties is high, but there is still a core of consultations that don’t seem to work. How do we make patients change their unhealthy behaviours? What can we do with consultations that repeatedly seem stuck on the same issues: the depressed patient who doesn’t get better, the diabetic …

Background: It has previously been reported that patients infected with Burkholderia cenocepacia (genomovar III) before lung transplantation have a poorer outcome than those with other B cepacia complex infections. Methods: An extensive study was conducted to determine the prevalence and clonality of B cepacia complex genomovars isolated from patients referred for transplant assessment between 1989 to the present and, where appropriate, whether strain type was related to transplant outcome. Results: Isolates from 29 patients were identified as B cepacia complex organisms by molecular analysis. Thirteen patients (45%) were infected with the highly transmissible ET-12 strain of B cenocepacia recA lineage III-A, while all remaining patients were infected with genetically unique B cenocepacia , B multivorans , and B vietnamiensis strains. All previously reported deaths following transplantation were associated with ET-12 infection. Conclusions: The ET-12 strain is the predominant cause of B cenocepacia infections in patients with cystic fibrosis referred to our pulmonary transplant centre and is associated with poor transplant outcomes using standard treatment regimens.

The anaerobic skin commensalPropionibacterium acnesis an underestimated cause of human infections and clinical conditions. Previous studies have suggested a role for the bacterium in lumbar disc herniation and infection. To further investigate this, five biopsy samples were surgically excised from each of 64 patients with lumbar disc herniation.P. acnesand other bacteria were detected by anaerobic culture, followed by biochemical and PCR-based identification. In total, 24/64 (38%) patients had evidence ofP. acnesin their excised herniated disc tissue. UsingrecAand mAb typing methods, 52% of the isolates were type II (50% of culture-positive patients), while type IA strains accounted for 28% of isolates (42% patients). Type III (11% isolates; 21% patients) and type IB strains (9% isolates; 17% patients) were detected less frequently. The MIC values for all isolates were lowest for amoxicillin, ciprofloxacin, erythromycin, rifampicin, tetracycline, and vancomycin (≤1mg/L). The MIC for fusidic acid was 1-2 mg/L. The MIC for trimethoprim and gentamicin was 2 to ≥4 mg/L. The demonstration that type II and III strains, which are not frequently recovered from skin, predominated within our isolate collection (63%) suggests that the role ofP. acnesin lumbar disc herniation should not be readily dismissed.

Propionibacterium acnes , a non-spore-forming, anaerobic Gram-positive bacterium, is most notably recognized for its association with acne vulgaris (I. Kurokawa et al., Exp. Dermatol. 18:821–832, 2009). We now present the draft genome sequence of an antibiotic-resistant P. acnes strain, PRP-38, isolated from an acne patient in the United Kingdom and belonging to the novel type IC cluster.

Propionibacterium acnes is an anaerobic Gram-positive bacterium that has been linked to a wide range of opportunistic human infections and conditions, most notably acne vulgaris (I. Kurokawa et al., Exp. Dermatol. 18:821–832, 2009). We now present the whole-genome sequences of three P. acnes strains from the type IA 2 cluster which were recovered from ophthalmic infections (A. McDowell et al., Microbiology 157:1990–2003, 2011).

Sleep problems can have a major impact on cognitive function, particularly in older adults who are also more likely to have a clinically diagnosed cognitive impairment. While several approved methods for sleep disturbance exist, most are not suitable for use within the abovementioned population. Often the reason for this is due to the symptoms associated with cognitive impairment or for the invasiveness of some sleep profiling methods. Developed from current sleep actigraphy techniques, this paper presents a non-contact alternative method for sleep profiling that is deemed to be more suitable for long term monitoring in the older population than current clinically approved techniques. This first evaluation has been conducted with a young control group in order to validate the approach. The results have demonstrated that based on the approach a random forest classifier using features calculated from optimally placed static accelerometers can produce a sleep/wake classification accuracy of 92%.

Propionibacterium acnes is an anaerobic Gram-positive bacterium that forms part of the normal human cutaneous microbiota and is thought to play a central role in acne vulgaris, a chronic inflammatory disease of the pilosebaceous unit (I. Kurokawa et al., Exp. Dermatol. 18: 821-832, 2009). Here we present the whole genome sequence of P. acnes type IB strain 6609, which was recovered from a skin sample from a woman with no recorded acne history and is thus considered a nonpathogenic strain (I. Nagy, Microbes Infect. 8: 2195-2205, 2006).

We have developed a novel multilocus sequence typing (MLST) scheme and database (http://pubmlst.org/pacnes/) for Propionibacterium acnes based on the analysis of seven core housekeeping genes. The scheme, which was validated against previously described antibody, single locus and random amplification of polymorphic DNA typing methods, displayed excellent resolution and differentiated 123 isolates into 37 sequence types (STs). An overall clonal population structure was detected with six eBURST groups representing the major clades I, II and III, along with two singletons. Two highly successful and global clonal lineages, ST6 (type IA) and ST10 (type IB1), representing 64 % of this current MLST isolate collection were identified. The ST6 clone and closely related single locus variants, which comprise a large clonal complex CC6, dominated isolates from patients with acne, and were also significantly associated with ophthalmic infections. Our data therefore support an association between acne and P. acnes strains from the type IA cluster and highlight the role of a widely disseminated clonal genotype in this condition. Characterization of type I cell surface-associated antigens that are not detected in ST10 or strains of type II and III identified two dermatan-sulphate-binding proteins with putative phase/antigenic variation signatures. We propose that the expression of these proteins by type IA organisms contributes to their role in the pathophysiology of acne and helps explain the recurrent nature of the disease. The MLST scheme and database described in this study should provide a valuable platform for future epidemiological and evolutionary studies of P. acnes.

Although two phenotypes of the opportunistic pathogen Propionibacterium acnes (types I and II) have been described, epidemiological investigations of their roles in different infections have not been widely reported. Using immunofluorescence microscopy with monoclonal antibodies (MAbs) QUBPa1 and QUBPa2, specific for types I and II, respectively, we investigated the prevalences of the two types among 132 P. acnes isolates. Analysis of isolates from failed prosthetic hip implants ( n = 40) revealed approximately equal numbers of type I and II organisms. Isolates from failed prosthetic hip-associated bone ( n = 6) and tissue ( n = 38) samples, as well as isolates from acne ( n = 22), dental infections ( n = 8), and skin removed during surgical incision ( n = 18) were predominately of type I. A total of 11 (8%) isolates showed atypical MAb labeling and could not be conclusively identified. Phylogenetic analysis of P. acnes by nucleotide sequencing revealed the 16S rRNA gene to be highly conserved between types I and II. In contrast, sequence analysis of recA and a putative hemolysin gene ( tly ) revealed significantly greater type-specific polymorphisms that corresponded to phylogenetically distinct cluster groups. All 11 isolates with atypical MAb labeling were identified as type I by sequencing. Within the recA and tly phylogenetic trees, nine of these isolates formed a cluster distinct from other type I organisms, suggesting a further phylogenetic subdivision within type I. Our study therefore demonstrates that the phenotypic differences between P. acnes types I and II reflect deeper differences in their phylogeny. Furthermore, nucleotide sequencing provides an accurate method for identifying the type status of P. acnes isolates.

We recently described a sonication technique for the diagnosis of prosthetic knee and hip infections. We compared periprosthetic tissue culture to implant sonication followed by sonicate fluid culture for the diagnosis of prosthetic shoulder infection. One hundred thirty-six patients undergoing arthroplasty revision or resection were studied; 33 had definite prosthetic shoulder infections and 2 had probable prosthetic shoulder infections. Sonicate fluid culture was more sensitive than periprosthetic tissue culture for the detection of definite prosthetic shoulder infection (66.7 and 54.5%, respectively; P = 0.046). The specificities were similar (98.0% and 95.1%, respectively; P = 0.26). Propionibacterium acnes was the commonest species detected among culture-positive definite prosthetic shoulder infection cases by periprosthetic tissue culture (38.9%) and sonicate fluid culture (40.9%). All subjects from whom P. acnes was isolated from sonicate fluid were male. We conclude that sonicate fluid culture is useful for the diagnosis of prosthetic shoulder infection.

Propionibacterium sp. is commonly isolated in association with orthopedic implants, either as a pathogen or a colonizer. Microbial characteristics that indicate whether the isolated species is a likely cause of orthopedic implant infection versus a colonizing agent would be clinically useful. We performed a prospective trial to determine the species of Propionibacterium and the phylotype (IA, IB, II, III) of Propionibacterium acnes isolated from the surface of removed orthopedic implants, and we correlated these findings with the presence or absence of infection. P. acnes represented 61 of 62 isolates. P. acnes type I was more commonly isolated than was type II (62% versus 38%, respectively), whether associated with infection or not. P. acnes type III was not detected. There was no clear association between types I and II P. acnes and infection or colonization of failed orthopedic implants (P = 0.75), however type IB strains were more frequently isolated than type IA from infected prosthese.

Acne is a common skin disorder of the pilosebaceous unit. In addition to genetic, hormonal and environmental factors, abnormal colonization by Propionibacterium acnes has been implicated in the occurrence of acne via the induction of inflammatory mediators. To gain more insight into the role that sebocytes play in the innate immune response of the skin, particularly in acne, we compared the antimicrobial peptide and proinflammatory cytokine/chemokine expression at mRNA and protein levels, as well as the viability and differentiation of SZ95 sebocytes in response to co-culture with representative isolates of P. acnes type IA and type IB as well as Escherichia coli-derived lipopolysaccharide (LPS). We found that, in vitro, P. acnes type IA and IB isolates and LPS induced human beta-defensin-2 and proinflammatory cytokine/chemokine expression, and influenced sebocyte viability and differentiation. Our results provide evidence that sebocytes are capable of producing proinflammatory cytokines/chemokines and antimicrobial peptides, which may have a role in acne pathogenesis. Furthermore, since P. acnes types IA and IB differentially affect both the differentiation and viability of sebocytes, our data demonstrate that different strains of P. acnes vary in their capacity to stimulate an inflammatory response within the pilosebaceous follicle.

Authors of previous studies have found that culture-based detection of prosthetic hip infections can be improved by adherence to strict anaerobic protocols and mild ultrasonication of retrieved prostheses to remove bacteria growing as an adherent biofilm. Furthermore, direct analyses of sonicate samples by immunofluorescence microscopy (with genus-specific and species-specific monoclonal antibodies or polyclonal antisera) produces significantly greater and more rapid detection rates for infection compared with culture alone. Despite its diagnostic value within a research setting, the practical advantages and limitations of immunofluorescence microscopy for a busy diagnostic laboratory needs consideration. In addition, the method must be evaluated against other nonculture-based techniques that may prove more appropriate. In this review, we describe how immunofluorescence microscopy presents an attractive and reliable method for routine detection of prosthetic hip biofilms compared, primarily, to polymerase chain reaction, although other nonculture methods of diagnosis based on fluorescent in situ hybridization and serologic analysis are considered. We describe how the immunofluorescence microscopy technique is robust, relatively simple and, in contrast to broad-range 16S recombinant-deoxyribonucleic-acid-based polymerase chain reaction, does not require a separate clean-room facility or strict adherence to aseptic techniques. More importantly we highlight how dislodged biofilm, which appears as large aggregates of bacteria, easily can be identified from skin contaminants that occur as single cells or small aggregates of only a few cells.

Analysis of the draft genome sequence of the opportunistic pathogenPropionibacterium acnestype strain NCTC 737 (=ATCC 6919) revealed five genes with sequence identity to the co-haemolytic Christie–Atkins–Munch-Peterson (CAMP) factor ofStreptococcus agalactiae. The predicted molecular masses for the expressed proteins ranged from 28 to 30 kDa. The genes were present in each of the three recently identifiedrecA-based phylogenetic groupings ofP. acnes(IA, IB and II), as assessed by PCR amplification. Conserved differences in CAMP factor gene sequences between these three groups were also consistent with their previous phylogenetic designations. All type IA, IB and II isolates were positive for the co-haemolytic reaction on sheep blood agar. Immunoblotting and silver staining of SDS-PAGE gels, however, revealed differential protein expression of CAMP factors amongst the different groups. Type IB and II isolates produced an abundance of CAMP factor 1, detectable by specific antibody labelling and silver staining of SDS-PAGE gels. In contrast, abundant CAMP factor production was lacking in type IA isolates, although larger amounts of CAMP factor 2 were detectable by immunoblotting compared with type II isolates. While the potential role of the abundant CAMP factor 1 in host colonization or virulence remains to be determined, it should be noted that the type strain ofP. acnesused in much of the published literature is a type IA isolate and is, therefore, lacking in this attribute.

Studies of the prevalence of Burkholderia cepacia complex species amongst cystic fibrosis (CF) patients in different geographical regions, and the association between cross-infection and putative transmissibility markers, will further our understanding of these organisms and help to address infection-control issues. In this study, B. cepacia complex isolates from CF patients in different regions of Europe were analysed. Isolates were examined for B. cepacia complex species and putative transmissibility markers [cable pilin subunit gene (cblA) and the B. cepacia epidemic strain marker (BCESM)]. Sporadic and cross-infective strains were identified by random amplification of polymorphic DNA (RAPD). In total, 79 % of patients were infected with Burkholderia cenocepacia (genomovar III), 18 % with Burkholderia multivorans (genomovar II) and less than 5 % of patients with B. cepacia (genomovar I), Burkholderia stabilis (genomovar IV) or Burkholderia vietnamiensis (genomovar V). The cblA and BCESM transmissibility markers were only detected in strains of B. cenocepacia. The BCESM was a more sensitive marker for transmissible B. cenocepacia strains than cblA, although sporadic B. cenocepacia strains containing the BCESM, but lacking cblA, were also observed. Furthermore, clusters of cross-infection with transmissibility marker-negative strains of B. multivorans were identified. In conclusion, B. cenocepacia was the greatest cause of cross-infection, and the most widely distributed B. cepacia complex species, within these CF populations. However, cross-infection was not exclusive to B. cenocepacia and cblA and the BCESM were not absolute markers for transmissible B. cenocepacia, or other B. cepacia complex strains. It is therefore suggested that CF centres cohort patients based on the presence or absence of B. cepacia complex infection and not on the basis of transmissibility marker-positive B. cenocepacia as previously suggested.

Oxidised low-density lipoprotein (LDL) has physicochemical properties that are pivotal in atherosclerotic plaque formation. As a consequence, antioxidant regimens may prove an important therapy in the prevention and treatment of cardiovascular disease. Since oxidised LDL is immunogenic, the aims of our study were to measure serum IgG titres to malondialdehyde-modified LDL (MDA-LDL) in patients with coronary artery disease (CAD) and control subjects and assess their potential as a clinical marker for coronary atherosclerosis and, consequently, antioxidant intervention. Serum IgG titres to MDA-LDL were measured in patients with angiographically confirmed CAD (n = 40) and aged-matched controls (n = 40) by enzyme-linked immunosorbant assay (ELISA). Titres were calculated and expressed as both the difference and the ratio of blanked absorbance units (AU) for IgG binding to MDA-LDL and native LDL. For the control population, median IgG titres were 0.26 AU (interquartile range (IQR) 0.20–0.46 AU) and 5.34 (IQR 3.40–8.58), respectively, while the patient population had median IgG titres of 0.30 AU (IQR 0.20–0.47 AU) and 5.08 (IQR 3.30–9.66), respectively. Both sets of calculated titre values were not significantly different between the two groups (P = 0.60 and 0.82, respectively). In conclusion, this study could not establish any significant association between circulating autoantibody titres to MDA-LDL and coronary atherosclerosis. Therefore, the diagnostic value of autoantibodies to oxidised LDL remains unclear.

We report on the first synthesis, kinetic evaluation and application of novel substrate-derived inhibitors against the Staphylococcus aureus cysteine protease-transpeptidase, sortase (staphylococcal surface protein sorting A, SrtA). The peptidyl-diazomethane and peptidyl-chloromethane analogues, Cbz (benzyloxycarbonyl)-Leu-Pro-Ala-Thr-CHN(2) (I) and Cbz-Leu-Pro-Ala-Thr-CH(2)Cl (II) respectively were found to act as time-dependent irreversible inhibitors of recombinant sortase (SrtA(DeltaN)). The peptidyl-chloromethane analogue (II) was the most powerful with an inhibitor specificity constant (k(i)/K(i)) of 5.3x10(4) M(-1).min(-1), approx. 2-fold greater than that determined for the peptidyl-diazomethane (I). Additionally, using Western-blot analysis, we have been able to demonstrate that a biotinylated version of the peptidyl-diazomethane analogue, biotin-Ahx (aminohexanoyl)-Leu-Pro-Ala-Thr-CHN(2) (III), can be used as an affinity label to detect the presence of wild-type SrtA in crude cell lysates prepared from S. aureus.

Burkholderia cepacia is a group of organisms that comprises seven genotypically distinct species (B cepacia genomovars I-VII), which are collectively known as the B cepacia complex. Preoperative infection with B cepacia is associated with a poor prognosis in lung transplant recipients with cystic fibrosis. Many centres do not, therefore, offer transplants to these individuals. Our aim was to ascertain whether or not post-transplant mortality is affected by pretransplant genomovar status. We studied archived isolates with PCR-based methods, and recorded excessive mortality in patients infected with B cepacia genomovar III, but not in those infected with other genomovars.

PCR amplification of the recA gene followed by restriction fragment length polymorphism (RFLP) analysis was investigated for the rapid detection and identification of Burkholderia cepacia complex genomovars directly from sputum. Successful amplification of the B . cepacia complex recA gene from cystic fibrosis (CF) patient sputum samples containing B . cepacia genomovar I, Burkholderia multivorans , B . cepacia genomovar III, Burkholderia stabilis , and Burkholderia vietnamiensis was demonstrated. In addition, the genomovar identifications determined directly from sputum were the same as those obtained after selective culturing. Sensitivity experiments revealed that recA -based PCR could reliably detect B . cepacia complex organisms to concentrations of 10 6 CFU g of sputum −1 . To fully assess the diagnostic value of the method, sputum samples from 100 CF patients were screened for B . cepacia complex infection by selective culturing and recA -based PCR. Selective culturing identified 19 samples with presumptive B . cepacia complex infection, which was corroborated by phenotypic analyses. Of the culture-positive sputum samples, 17 were also detected directly by recA -based PCR, while 2 samples were negative. The isolates cultured from both recA -negative sputum samples were subsequently identified as Burkholderia gladioli . RFLP analysis of the recA amplicons revealed 2 patients (12%) infected with B . multivorans , 11 patients (65%) infected with B . cepacia genomovar III-A, and 4 patients (23%) infected with B . cepacia genomovar III-B. These results demonstrate the potential of recA -based PCR-RFLP analysis for the rapid detection and identification of B . cepacia complex genomovars directly from sputum. Where the sensitivity of the assay proves a limitation, sputum samples can be analyzed by selective culturing followed by recA -based analysis of the isolate.

We have developed matrix metalloprotease (MMP) inhibitors based on synthetic peptides incorporating a non-cleavable peptide-bond isostere at the site of the putative scissile bond. These inhibitors, N-α-phthaloyl-Gly-Ψ(CO-CH2)-Leu-Tyr-Ala-NH2 (Pht-G-CH2-LYA-NH2) and N-α-phthaloyl-Gly-Ψ(CO-CH2)-Leu-Tic-Ala-NH2 (Pht-G-CH2-LTcA-NH2) were kinetically evaluated against the type IV collagenases, gelatinase A (MMP-2) and B (MMP-9), and compared with an exactly analogous chelating-based inhibitor, N-α-mercaptoacetyl-Leu-Tyr-Ala-NH2 (HSCH2CO-LYA-NH2). The peptide inhibitors were also tested for their anti-invasive effects on breast carcinoma cell lines using a modification of the Boyden chamber assay. Gelatin zymography was utilized to identify gelatinolytic activities present in media removed from cultured breast cancer cells. Of the two N-α-phthalimidomethyl-ketomethylene peptide-based inhibitors, Pht-G-CH2-LYA-NH2 proved the more effective inhibitor of MMP-2 and MMP-9 (Ki 34.27 and 45.75 μm, respectively). However, when tested against two breast cancer cell lines, T47D and MDA-MB-231, both inhibitors were able to effectively reduce tumour cell invasion through a type IV collagen matrix by up to 91.2%. Of particular interest was the observation that Pht-G-CH2-LYA-NH2 was the most potent inhibitor of invasion by the highly aggressive MDA-MB-231 cells, despite the cells' relative lack of active secreted metalloprotease activity. The results obtained from this kinetic and anti-invasive analysis of the new inhibitors suggest that compounds incorporating the N-α-phthalimidomethyl-ketomethylene peptide-bond isostere may have potential for development as new agents with anti-metastatic properties.

A series of electrophilic glutamine analogues based on 6-diazo-5-oxo-norleucine has been prepared, using novel synthetic routes, and evaluated as inhibitors of Escherichia coli glucosamine synthetase. The gamma-dimethylsulphonium salt analogue of glutamine was found to be one of the most potent inactivators of this enzyme yet reported, with an apparent second order rate constant (k2/Ki) of 3.5 x 10(5) M(-1) min(-1).

We tested the hypothesis that male southern green stink bugs, Nezara viridula (L.), use substrate-borne songs to locate females. We recorded the responses of bugs on plants to the vibrations caused by a prerecorded female song and by an artificial sound. The female song caused males to walk, to respond with the calling and courtship songs and to approach the source of the song with characteristic search behaviour at junctions between branches on the plants. At a junction, a searching male stopped, stretched his legs and antennae and compared the vibratory signals on the two branches, with different combinations of legs and antennae. The males then left the junction and approached the source of the vibration. Males located the loudspeaker significantly more frequently in the presence than in the absence of vibratory stimuli on cyperus, Cyperus alternifolius L., and beans, Phaseolus vulgaris L. Vibrational directionality was also elicited by artificial pure tones whose spectral and temporal parameters were similar to those of natural female song. Females showed no reaction to vibratory stimulation and no vibrational directionality. We discuss possible mechanisms underlying vibrational directionality in the light of expected signal changes during transmission through plants. Copyright 1999 The Association for the Study of Animal Behaviour.

Based upon current sleep actigraphy techniques, this paper discusses an alternative non-contact method of sleep profiling that is potentially more suitable for long term monitoring than current clinically approved techniques. The passive sleep actigraphy PSA platform presented here utilises strategically positioned accelerometers fixed on a mattress to quantify the recorded movements of a bed occupant. In this work, data captured from a young control group is decomposed into gravitational and inertial components. These components are then translated into activity counts using numerous quantification modalities and feature extraction techniques to isolate the most discriminant attributes for optimal sleep/wake classification. These attributes were then input into a random forest classifier to determine the sleep/wake state of each subject based on their recoded actigraphy data with an accuracy of 89%. The findings suggest that the PSA platform is a potentially viable method of non-contact sleep profiling hence supporting further research into this approach.

We investigated the ability of secreted bacterial proteinases from three pathogens [Burkholderia multivorans (Bm), Burkholderia cenocepacia (Bc), and Pseudomonas aeruginosa (Pa)] involved in chronic bacterial infections in cystic fibrosis to degrade various host defence-related molecules. These included two endogenous proteinase inhibitors, secretory leukocyte proteinase inhibitor (rhSLPI) and alpha-1 antitrypsin (AAT); two relevant immunoglobulins, secretory IgA (sIgA) and IgG, and two proteins important in innate immunity, lactoferrin and lysozyme. Host defence-related molecules were co-incubated with cell-free supernatants from 48 hour biofilm cultures, grown on mucin-coated microplates, from all three pathogens under investigation (six isolates from each species). No degradation of AAT, sIgA, IgG, and lactoferrin was observed for any of the organisms. Of the 18 isolates tested, only one demonstrated the ability to degrade lysozyme. This was an environmental isolate of Pa which was included as a comparison for the predominantly clinically relevant isolates used in the study. In contrast, all isolates of Bm (n = 4) and Pa (n = 4) were able to degrade rhSLPI however, out of five bacterial isolates tested for Bc only two demonstrated a limited ability to degrade the molecule with >95% of the protein band still remaining intact at the end of the experiment. This study demonstrates that the majority of the host defence molecules investigated are resistant to degradation by bacterial proteinases from Bm, Bc and Pa when grown as a biofilm. However, rhSLPI was vulnerable to significant degradation which could result in aberrant serine proteolysis in regions of the lungs containing biofilm growth.

Background Patients with cystic fibrosis [CF] have frequent pulmonary exacerbations associated with the isolation of bacterial organisms from sputum samples. It is not clear however, if there are differences in the types of additional organisms isolated from patients who are infected with Burkholderia cepacia complex [BCC] or Pseudomonas aerugionsa [PA] in comparison to those who are not infected with either of these organisms [NI].