Development and validation of a reference marker for identification of aerobic and anaerobic bacteria associated with diabetes chronic wound ulcers using PCR denaturing gradient gel electrophoresis

Introduction: Diabetes chronic wounds consist with a diverse microbial community and unculturablespecies may be highly prevalent.Objectives: This study aimed to establish a bacterial reference marker consisting of a group ofchronic wound related bacteria, using polymerase chain reaction-denaturing gradient gelelectrophoresis (PCR-DGGE) for profiling of bacteria in diabetes chronic wound infections.Methods: DNA was extracted from the known wound bacterial strains. PCR–DGGE was performedusing eubacterial specific primers targeting V2-V3 region of 16S rDNA. DGGE was performed usinga 30-55% denaturing gradient. Migration position of each organism was detected on DGGE gel andimportant organisms were selected. Equal volume from PCR products of each selected organism wasmixed, diluted with gel loading dye in 1:1.5 ratio and used for all DGGE gels. The ladder was thensubjected to species identification of fifteen tissue debridement specimens obtained from diabeteschronic wound ulcers. The identification efficacy was tested by sequencing.Results: DNA of bacterial pathogens which showed different migration distances on the gel werecombined and used as a reference panel. This bacterial ladder consisted of eleven different bacterialspecies including Bacteroides sp., S. aureus, Acineto bacter sp., P. aeruginosa, Streptococcus Group Aand Group B sp., E. faecalis, Providencia sp., Veillonella sp., E .coli and Enterobacter sp. Accordingto the reference panel, Pseudomonas species were abundant. Further the results were confirmed bysequencing.Conclusion: Reference marker allows comparative analysis of DGGE patterns and can be used as atool for presumptive identification of polymicrobial microbiota in chronic wound infections

Cytomegalovirus, Epstein–Barr virus and risk of breast cancer before age 40 years: a case–control study

We investigated whether there is an association between cytomegalovirus (CMV) and Epstein-Barr virus (EBV) IgG levels and risk of breast cancer before age 40 years. CMV and EBV IgG levels were measured in stored plasma from 208 women with breast cancer and 169 controls who participated in the Australian Breast Cancer Family Study (ABCFS), a population-based case-control study. CMV and EBV IgG values were measured in units of optical density (OD). Cases and controls did not differ in seropositivity for CMV (59 and 57% respectively; P=0.8) or EBV (97 and 96% respectively; P=0.7). In seropositive women, mean IgG values were higher in cases than controls for CMV (1.20 vs 0.98 OD, P=0.005) but not for EBV (2.65 vs 2.57 OD, P=0.5). The adjusted odds ratios per OD unit were 1.46 (95% CI 1.06-2.03) for CMV IgG and 1.11 (0.93-1.33) for EBV IgG. The higher mean CMV IgG levels found in women with breast cancer could be the result of a more recent infection with CMV, and may mean that late exposure to CMV is a risk factor for breast cancer

Linking the sectoral employment structure and household poverty in the United Kingdom

Structural changes in the labour markets of developed economies, and changes in their institutional characteristics, have led to growing unease about the nature of low-paid employment. Related concerns have been expressed about the persistence of low-pay, the fragmentation of work and the growth of under-employment. While all these factors have potential implications for individuals’ earnings, less is known about the connection between labour market change, patterns of sectoral growth and decline and household poverty outcomes. This article shows distinct patterns of poverty outcomes by sector of employment, after controlling for other factors. However, household characteristics, in particular the presence of a second earner, do strongly mitigate the poverty risk. Overall, the findings demonstrate that policymakers need to develop a coherent policy towards poverty that recognises the nature of jobs growth and the distribution of ‘good jobs’ across households

The geography of wage inequality in British cities

There is widespread concern about the scale and implications of urban inequality in Great Britain, but little evidence on which cities are the most unequal and why. This paper investigates patterns of wage inequality in 60 British cities. It has two principal goals: (1) to describe which cities are most unequal and (2) to assess the important determinants of inequality. The results show a distinct geography of wage inequality, the most unequal cities tend to be affluent and located in parts of the Greater South East of England. A central determinant of these patterns is the geography of highly skilled workers. Because of this, the geography of urban wage inequality reflects the geography of affluence more generally

Growth inhibition of oral mutans streptococci and candida by commercial probiotic lactobacilli - an in vitro study

<p>Abstract</p> <p>Background</p> <p>Probiotic bacteria are suggested to play a role in the maintenance of oral health. Such health promoting bacteria are added to different commercial probiotic products. The aim of the study was to investigate the ability of a selection of lactobacilli strains, used in commercially available probiotic products, to inhibit growth of oral mutans streptococci and <it>C. albicans in vitro</it>.</p> <p>Methods</p> <p>Eight probiotic lactobacilli strains were tested for growth inhibition on three reference strains and two clinical isolates of mutans streptococci as well as two reference strains and three clinical isolates of <it>Candida albicans </it>with an agar overlay method.</p> <p>Results</p> <p>At concentrations ranging from 10⁹to 10⁵CFU/ml, all lactobacilli strains inhibited the growth of the mutans streptococci completely with the exception of <it>L. acidophilus </it>La5 that executed only a slight inhibition of some strains at concentrations corresponding to 10⁷and 10⁵CFU/ml. At the lowest cell concentration (10³CFU/ml), only <it>L. plantarum </it>299v and <it>L. plantarum </it>931 displayed a total growth inhibition while a slight inhibition was seen for all five mutans streptococci strains by <it>L. rhamnosus </it>LB21, <it>L. paracasei </it>F19, <it>L. reuteri </it>PTA 5289 and <it>L. reuteri </it>ATCC 55730. All the tested lactobacilli strains reduced candida growth but the effect was generally weaker than for mutans streptococci. The two <it>L. plantarum </it>strains and <it>L. reuteri </it>ATCC 55730 displayed the strongest inhibition on <it>Candida albicans</it>. No significant differences were observed between the reference strains and the clinical isolates.</p> <p>Conclusion</p> <p>The selected probiotic strains showed a significant but somewhat varying ability to inhibit growth of oral mutans streptococci and <it>Candida albicans in vitro</it>.</p

The Maltase Involved in Starch Metabolism in Barley Endosperm Is Encoded by a Single Gene

During germination and early seedling growth of barley (Hordeum vulgare), maltase is responsible for the conversion of maltose produced by starch degradation in the endosperm to glucose for seedling growth. Despite the potential relevance of this enzyme for malting and the production of alcoholic beverages, neither the nature nor the role of maltase is fully understood. Although only one gene encoding maltase has been identified with certainty, there is evidence for the existence of other genes and for multiple forms of the enzyme. It has been proposed that maltase may be involved directly in starch granule degradation as well as in maltose hydrolysis. The aim of our work was to discover the nature of maltase in barley endosperm. We used ion exchange chromatography to fractionate maltase activity from endosperm of young seedlings, and we partially purified activity for protein identification. We compared maltase activity in wild-type barley and transgenic lines with reduced expression of the previously-characterised maltase gene Agl97, and we used genomic and transcriptomic information to search for further maltase genes. We show that all of the maltase activity in the barley endosperm can be accounted for by a single gene, Agl97. Multiple forms of the enzyme most likely arise from proteolysis and other post-translational modifications

Oral biofilm models for mechanical plaque removal

In vitro plaque removal studies require biofilm models that resemble in vivo dental plaque. Here, we compare contact and non-contact removal of single and dual-species biofilms as well as of biofilms grown from human whole saliva in vitro using different biofilm models. Bacteria were adhered to a salivary pellicle for 2 h or grown after adhesion for 16 h, after which, their removal was evaluated. In a contact mode, no differences were observed between the manual, rotating, or sonic brushing; and removal was on average 39%, 84%, and 95% for Streptococcus mutans, Streptococcus oralis, and Actinomyces naeslundii, respectively, and 90% and 54% for the dual- and multi-species biofilms, respectively. However, in a non-contact mode, rotating and sonic brushes still removed considerable numbers of bacteria (24–40%), while the manual brush as a control (5–11%) did not. Single A. naeslundii and dual-species (A. naeslundii and S. oralis) biofilms were more difficult to remove after 16 h growth than after 2 h adhesion (on average, 62% and 93% for 16- and 2-h-old biofilms, respectively), while in contrast, biofilms grown from whole saliva were easier to remove (97% after 16 h and 54% after 2 h of growth). Considering the strong adhesion of dual-species biofilms and their easier more reproducible growth compared with biofilms grown from whole saliva, dual-species biofilms of A. naeslundii and S. oralis are suggested to be preferred for use in mechanical plaque removal studies in vitro