A systems biology approach to dissection of the effects of small bicyclic peptidomimetics on a panel of Saccharomyces cerevisiae mutants

In recent years, an approach called “chemical genetics” has been adopted in drug research to discover and validate new targets and to identify and optimize leads by high throughput screening. In this work, we tested the ability of a library of small peptidomimetics to induce phenotypic effects with functional implications on a panel of strains of the budding yeast Saccharomyces cerevisiae, both wild type and mutants, for respiratory function and multidrug resistance. Further elucidation of the function of these peptidomimetics was assessed by testing the effects of the compound with the most prominent inhibitory activity, 089, on gene expression using DNA microarrays. Pathway analysis showed the involvement of such a molecule in inducing oxidative damage through alterations in mitochondrial functions. Transcriptional experiments were confirmed by increased levels of ROS and activation of mitochondrial membrane potential. Our results demonstrate the influence of a functional HAP1 gene in the performance of S. cerevisiae as a model system

Dietary Nitrate: Effects on the health of weaning pigs and Antimicrobial activity on seven probiotic Bifidobacterium spp. strains

The potential role of nitrite as an antimicrobial substance in the stomach may be of some importance in the ecology of the gastrointestinal tract and in host physiology. It has been shown that nitrite, under the acidic conditions of the stomach, may kill gut pathogens like Salmonella enteritidis, Escherichia coli, Salmonella typhimurium, and Yersinia enterocolitica, whereas acid alone has only a bacteriostatic effect. An in vivo study was conducted in order to assess the effects of dietary nitrate on microbiota and on the health of the gut (particularly in the stomach and small intestine). 96 weaning pigs were fed a diet containing high nitrate levels (15 mg and 150 mg) and then challenged with Salmonella enterica serovar typhimurium. Differences in composition of the gut microbiota were assessed by analysing samples from the pigs: To date analysis of 48 pigs has been completed.. Preliminary results demonstrated no effect on the population densities of microbial groups either from the challenge or from nitrate intake. However, increasing the time from challenge decreased either the counts of LAB in the stomach and jejunum or of clostridia in the stomach. Bifidobacteria also decreased in the stomach contents as nitrate supplementation increased. Supplementing the feedstuff with high dietary nitrate intake and then challenging with Salmonella did not affect the gastric pH or the degree of ulceration in the pigs. The synergistic bactericidal effects of pH, nitrite and thiocyanate on seven probiotic Bifidobacterium spp. strains were also investigated in an in vitro study. The results of the in vitro study demonstrated that an inhibitory effect exists on the seven probiotic bifidobacteria investigated with an exposure longer than 2 hours and pH values < 5.0. Addition of thiocyanate also increased the susceptibility of the tested strains. In this in vitro study, the most resistant strains at all conditions were B. animalis subsp. lactis Ra 18 and P32 and B. choerinum Su 877, Su 837 and Su 891

Transcription-associated breaks in Xeroderma Pigmentosum group D cells from patients with combined features of Xeroderma Pigmentosum and Cockayne Syndrome

Defects in the XPD gene can result in several clinical phenotypes, including xeroderma pigmentosum (XP), trichothiodystrophy, and, less frequently, the combined phenotype of XP and Cockayne syndrome (XP-D/CS). We previously showed that in cells from two XP-D/CS patients, breaks were introduced into cellular DNA on exposure to UV damage, but these breaks were not at the sites of the damage. In the present work, we show that three further XP-D/CS patients show the same peculiar breakage phenomenon. We show that these breaks can be visualized inside the cells by immunofluorescence using antibodies to either gamma-H2AX or poly-ADP-ribose and that they can be generated by the introduction of plasmids harboring methylation or oxidative damage as well as by UV photoproducts. Inhibition of RNA polymerase II transcription by four different inhibitors dramatically reduced the number of UV-induced breaks. Furthermore, the breaks were dependent on the nucleotide excision repair (NER) machinery. These data are consistent with our hypothesis that the NER machinery introduces the breaks at sites of transcription initiation. During transcription in UV-irradiated XP-D/CS cells, phosphorylation of the carboxy-terminal domain of RNA polymerase II occurred normally, but the elongating form of the polymerase remained blocked at lesions and was eventually degraded

Antimicrobial resistance, an update from the ward: Increased incidence of new potential pathogens and site of infection-specific antibacterial resistances

In order to monitor the spread of antimicrobial resistance, the European Union requires hospitals to be equipped with infection control centers. With this aim, we analyzed 1583 bacterial strains isolated from samples of different origin from patients with community-onset and nosocomial infections in a public tertiary University Hospital on the outskirts of Turin, Italy. Statistical analyses of the isolates (source, type) and their antimicrobial resistance (AMR) were performed. The survey revealed infections associated with bacterial species considered as not-commensal and not-pathogenic, hence potentially emerging as new threats for human health. Conversely to the general observation of nosocomial strains being more resistant to antibiotics compared to community-acquired strains, nosocomial strains isolated in this study were more resistant only to 1/42 tested antibiotics (tetracycline). By adopting an ecological approach, we observed that blood infections are associated with the broadest range of species compared to infections affecting other areas and we obtained clear indications on the antibiotics that should be preferred in the treatment of infections at specific body sites. Future investigations carried out on a larger geographical scale will clarify whether these indications are limited to the geographical region investigated over this study, or whether the same trends are visible at national or international level