Biofilm induced tolerance towards antimicrobial peptides

Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We address the question of how biofilm organization affects antibiotic susceptibility. We established Escherichia coli biofilms with differential structural organization due to the presence of IncF plasmids expressing altered forms of the transfer pili in two different biofilm model systems. The mature biofilms were subsequently treated with two antibiotics with different molecular targets, the peptide antibiotic colistin and the fluoroquinolone ciprofloxacin. The dynamics of microbial killing were monitored by viable count determination, and confocal laser microscopy. Strains forming structurally organized biofilms show an increased bacterial survival when challenged with colistin, compared to strains forming unstructured biofilms. The increased survival is due to genetically regulated tolerant subpopulation formation and not caused by a general biofilm property. No significant difference in survival was detected when the strains were challenged with ciprofloxacin. Our data show that biofilm formation confers increased colistin tolerance to cells within the biofilm structure, but the protection is conditional being dependent on the structural organization of the biofilm, and the induction of specific tolerance mechanisms.     

Alkylation damage in DNA and RNA--repair mechanisms and medical significance

Alkylation lesions in DNA and RNA result from endogenous compounds, environmental agents and alkylating drugs. Simple methylating agents, e.g. methylnitrosourea, tobacco-specific nitrosamines and drugs like temozolomide or streptozotocin, form adducts at N- and O-atoms in DNA bases. These lesions are mainly repaired by direct base repair, base excision repair, and to some extent by nucleotide excision repair (NER). The identified carcinogenicity of O(6)-methylguanine (O(6)-meG) is largely caused by its miscoding properties. Mutations from this lesion are prevented by O(6)-alkylG-DNA alkyltransferase (MGMT or AGT) that repairs the base in one step. However, the genotoxicity and cytotoxicity of O(6)-meG is mainly due to recognition of O(6)-meG/T (or C) mispairs by the mismatch repair system (MMR) and induction of futile repair cycles, eventually resulting in cytotoxic double-strand breaks. Therefore, inactivation of the MMR system in an AGT-defective background causes resistance to the killing effects of O(6)-alkylating agents, but not to the mutagenic effect. Bifunctional alkylating agents, such as chlorambucil or carmustine (BCNU), are commonly used anti-cancer drugs. DNA lesions caused by these agents are complex and require complex repair mechanisms. Thus, primary chloroethyl adducts at O(6)-G are repaired by AGT, while the secondary highly cytotoxic interstrand cross-links (ICLs) require nucleotide excision repair factors (e.g. XPF-ERCC1) for incision and homologous recombination to complete repair. Recently, Escherichia coli protein AlkB and human homologues were shown to be oxidative demethylases that repair cytotoxic 1-methyladenine (1-meA) and 3-methylcytosine (3-meC) residues. Numerous AlkB homologues are found in viruses, bacteria and eukaryotes, including eight human homologues (hABH1-8). These have distinct locations in subcellular compartments and their functions are only starting to become understood. Surprisingly, AlkB and hABH3 also repair RNA. An evaluation of the biological effects of environmental mutagens, as well as understanding the mechanism of action and resistance to alkylating drugs require a detailed understanding of DNA repair processes.     

Factors influencing the social acceptability of large carnivore behaviours

A survey on attitudes toward large carnivores was conducted in a representative sample of the Norwegian population (n = 3134). People were asked about the acceptability of carnivores living in remote wilderness, close to where people live, killing livestock, killing pets, or threatening humans. Large differences in acceptability appeared across the five situations. Wolves and bears were less acceptable than lynx and wolverines when observed close to where people live. Negative associations were found between acceptability and lack of personal control, economic loss, and respondents' age. Acceptability was higher among males than among females, and higher among urban than among rural residents. The results showed that general measures of attitudes alone toward large carnivores were of limited value in wildlife management. The situational and social specificity of these attitudes should be given more attention.     

Proteome profiles of mucosal immunoglobulin uptake in inflamed porcine gut

Acquisition of passive immunity by endocytosis of intact immunoglobulins (Ig) from colostrum is critical for prevention of intestinal and systemic diseases in neonatal mammals. We compared proteome patterns of healthy and inflamed gut tissues from pre-term piglets to investigate the effect of inflammation on acquisition of passive immunity. A clear difference in the two-dimensional gel electrophoresis protein patterns between healthy and inflamed intestinal tissues was observed, suggesting that inflamed tissues failed to absorb and transfer Ig from colostrum to epithelial cells. We have mapped and identified the Ig proteins that are taken up by healthy intestinal tissues, and found that isoforms of the IgA and IgG heavy chain and Ig kappa and lambda light chains were internalized. Our results indicate that colostrum protein uptake in the porcine gut is a selective process that is obstructed in inflamed pre-term gut.     

Prevalence of Edwards' syndrome. Clustering and seasonal variation?

Summary The incidence of Edwards' syndrome was found to be 1 per 4857 newborn children of 34000 consecutively newborn children in two Danish counties. Six of the 7 cases were born during the months of February through April.The incidence was high compared with the expected incidence of Edwards' syndrome of approximately 1 per 10000. This might be due to clustering in the area studied during the period 1967 to 1973.The finding of variations in incidence of children with Edwards' syndrome in different parts of the world, as well as the finding of seasonal variation in birth of such children, indicates that some of the etiological factors of nondisjunction of chromosome 18 are of an environmental nature.     

Traditional Ecological Knowledge of a Riverine Forest in Turkana, Kenya: Implications for Research and Management

The present study explores traditional ecological knowledge (TEK) of Turkana pastoralists and cultivators in the context of a riverine forest in northern Kenya. The Turkwel River and its floodplain sustain a thick forest, which is used for grazing and extraction of non-timber forest products. However, sedentarisation and agricultural expansion have resulted in localised clear-felling of trees, while river damming has altered the natural flow regime. A series of structured, semi-structured, and group interviews were combined with a botanical inventory in order to assess the relevance of TEK to ecological research and forest conservation. Turkana informants gave 102 vernacular names for the 113 woody species. Of these, 85% had a domestic or pastoral use among the 105 specific uses that were described. Ethnobotanical knowledge was relatively homogenous and not related to age, gender, or source of livelihood. The informants had in-depth knowledge of some key ecological processes. The conceived threats to forest survival were primarily cultivation and permanent settlements, while the effects of river damming and livestock grazing were disputed. A claimed decline in rainfall was confirmed by official data. There is strong evidence that TEK could be used to generate hypotheses for research and to design sustainable conservation strategies. A revised version of the indigenous system of tree management should be incorporated into the official forestry policy in order to resolve future conflicts between pastoralists and cultivators.