Recombinant yeast and human cells as screening tools to search for antibacterial agents targeting the transcription termination factor Rho

International audienceThe alarming issue of antibiotic resistance expansion requires a continuous search for new and efficient antibacterial agents. Here we describe the design of new tools to screen for target-specific inhibitors of the bacterial Rho factor directly inside eukaryotic cells. Rho factor is a global regulator of gene expression which is essential to most bacteria, especially Gram-negative. Since Rho has no functional or structural homolog in eukaryotes, it constitutes a valuable and well known bacterial target as evidenced by its inhibition by the natural antibiotic, Bicyclomycin. Our screening tools are based on perturbation of mRNA processing and packaging reactions in the nucleus of eukaryotic cells by the RNA-dependent helicase/translocase activity of bacterial Rho factor leading to a growth defect phenotype. In this approach, any compound that impedes Rho activity should restore growth to yeast or human cells expressing Rho protein, providing valuable means to screen for target-specific antibacterial agents within the environment of a eukaryotic cell. The yeast tool expressing E. coli Rho factor was validated using Bicyclomycin as the control antibacterial agent. The validation of the screening tool was further extended with a stable human cell line expressing Rho factor conditionally. Finally, we show that Rho factors from different bacterial pathogens can also be designed as yeast-based screening tools which can reveal subtle variations in the functional features of the proteins

Results of surgical and radiotherapeutic treatment of adenoid cystic carcinoma of the salivary glands

Surgical treatment in patients with rare adenoid cystic carcinoma (ACC) of the salivary glands is considered to be the state of the art. With respect to an additional radiotherapy there are different approaches regarding the type of radiotherapy and timing. In this study the overall survival and recurrence-free survival in 52 individuals with salivary gland ACC who were treated at the University Hospital in Essen and received irradiation with fast neutrons and photons (mixed beam technique) either A) immediately following surgical treatment or B) only after the appearance of local recurrence were compared. Group A (n = 28, first diagnosis, FD September 1991-September 2009) received adjuvant radiotherapy immediately postoperative, group B (n = 24, FD June 1979-November 2001) underwent primarily surgical tumor resection according to the treatment regimen at that time and were irradiated only on the appearance of a local recurrence. In comparison to group B, patients in group A showed a lower recurrence rate and a significantly longer local relapse-free survival. Group B, however, showed a significantly higher overall survival. The frequency of distant metastasis occurred equally in both groups but the onset of distant metastasis was significantly earlier in group A. In general, overall survival was negatively influenced by distant metastasis. The local recurrence rate was very high after primary surgical treatment only. The immediate adjuvant high-linear energy transfer (LET) radiotherapy reduced the local recurrence rates. Irradiation after the appearance of a recurrence had a positive influence on overall survival. Overall, definitive high-LET radiotherapy in the mixed beam technique enabled high local control rates both primarily postoperative and also locoregional recurrences

Modeling habitat preferences of Caspian kutum, Rutilus frisii kutum (Kamensky, 1901) (Actinopterygii, Cypriniformes) in the Caspian Sea

Predicting and modeling of habitat preferences of fish is a very important issue for aquatic management. Classification trees (CTs) were used to predict the habitat preferences of the Caspian kutum (Rutilus frisii kutum, hereafter kutum) in the southern Caspian Sea. The applied model was optimized with genetic algorithm (GA) and greedy stepwise (GS) to select the most explanatory variables for predicting the presence/absence of kutum. The suitability index was considered to determine the quality and suitability of fish habitat in the sea. The results of Paired Student's t tests showed that there was a significant difference between predictive performances of models before and after variable selection methods. Both optimizers improved the predictive power of CTs and resulted in a better understanding of CTs by making a selection of the sea characteristics that were used as inputs to the models. The results show that the effect of different seasons, sea depth, and photosyntheticaly active radiation were the main predictors affecting the habitat preferences of kutum in the Caspian Sea. Constructed trees in combination with GA and GS showed high capability when applied to predict the habitat preferences of this valuable commercial fish species. Determining the habitat needs of the target fish will enhance local fisheries performances and the long-term conservation planning of the fish to implement the ecosystem-based management in the Caspian Sea