The Dimerization Interface in VraR is Essential to Induction of the Cell Wall Stress Response in Staphylococcus aureus: A potential Druggable Target

Background Staphylococcus aureus remains a medical challenge in the treatment of bacterial infections. It has acquired resistance to commonly used antibiotics, and to those considered to be the last weapons in treating staphylococcal infections, such as vancomycin. Studies have revealed that S. aureus is capable of mounting a rapid response to antibiotics that target cell wall peptidoglycan biosynthesis, such as β-lactams and vancomycin. The two-component system VraSR has been linked to the coordination of this response. VraS is a histidine kinase that undergoes autophosphorylation in the presence of signals elicited upon cell wall damage and it then transfers its phosphoryl group to VraR. VraR is a response regulator protein that functions as a transcription factor. Phosphorylation of VraR leads to its dimerization, which is required for optimum binding to its target promoters. Two-component systems have been targeted for the development of antibacterial agents. Deletion of the vraS or vraR gene has been shown to re-sensitize S. aureus to β-lactams and vancomycin. Results In this study, we explored perturbation of the VraR phosphorylation-induced activation as a means to inhibit the VraSR-mediated signal transduction pathway. We show that dimerization of VraR is essential for the phosphorylation-induced activation of VraR. A single point mutation in the dimerization interface of VraR, in which Met13 was replaced by Ala, led to the inability of VraR to dimerize and to bind optimally to the target promoter. The consequences of these in vitro molecular deficiencies are equally dramatic in vivo. Complementation of a vraR deletion S. aureus strain with the vraRM13Ala mutant gene failed to induce the cell wall stress response. Conclusions This study highlights the potential of targeting the phosphorylation-induced dimerization of VraR to disrupt the S. aureus cell wall stress response and in turn to re-sensitize S. aureus to β-lactams and vancomycin.York University Librarie

Spontaneous thermal expansion of nematic elastomers

We study the monodomain (single-crystal) nematic elastomer materials, all side-chain siloxane polymers with the same mesogenic groups and crosslinking density, but differing in the type of crosslinking. Increasing the proportion of long di-functional segments of main-chain nematic polymer, acting as network crosslinking, results in dramatic changes in the uniaxial equilibrium thermal expansion on cooling from isotropic phase. At higher concentration of main chains their behaviour dominates the elastomer properties. At low concentration of main-chain material, we detect two distinct transitions at different temperatures, one attributed to the main-chain, the other to the side-chain component. The effective uniaxial anisotropy of nematic rubber, r(T) proportional to the effective nematic order parameter Q(T), is given by the average of the two components and thus reflects the two-transition nature of thermal expansion. The experimental data is compared with the theoretical model of ideal nematic elastomers; applications in high-amplitude thermal actuators are discussed in the end

Isolation and Molecular Detection of Gram Negative Bacteria Causing Urinary Tract Infection in Patients Referred to Shahrekord Hospitals, Iran

Background: Urinary Tract Infections (UTI), and their complications, cause serious health problems, which affect millions of people every year. Infections of the urinary tract are the second most common type of infection in the body and approximately 20% of women are especially prone to UTIs for reasons not yet well understood. Urinary Tract Infections in men are not as common as in women yet can be very serious when they do occur. Accurate identification of bacterial isolates is an essential task of the clinical microbiology laboratory. Objectives: The purpose of this study was to determine the incidence and variety of the causative microbial agents of UTIs in patients who had referred to a medical laboratory of Kashani and Hajar hospital in Shahrekord, Iran. Patients and Methods: In this cross-sectional study 147 urine samples of patients (urine test results were positive for UTIs) were examined during April to September 2013. A total of 147 urine samples of patients with clinical symptoms of UTI who had been referred to a medical laboratory of Kashani and Hajar hospital in Shahrekord (Iran), were collected and processed immediately for laboratory analysis. Results: Escherichia coli was identified as the most common causative agent of UTIs (51.70% of total isolates in both sexes), followed by Klebsiella pneumoniae (K. Pneumoniae) (16.32%). Frequency of Proteus spp., Acinetobacter spp., Entrobacter spp., Citrobacter spp., Pseudomonas aeruginosa (P. aeruginosa) and Providencia spp. was 10.88%, 6.12%, 5.44%, 4.08%, 3.40% and 2.04%, respectively. Statistical analysis by Fisher exact test showed that there was no significant relationship between the type of bacteria and gender (P > 0.05). Chi square test showed that there was no significant relationship between the type of bacteria and the use of catheter and age group (P > 0.05). However, there was a significant relationship between the type of bacteria and the history of hospitalization (P > 0.05). Conclusions: Our findings implied that a wide range of bacteria could be involved in creating urinary tract infection in patients referred to a medical laboratory of Kashani and Hajar hospital in Shahrekord, Iran. Regardless of age, sex and the use of catheter, a wide range of bacteria could be involved in urinary tract infections

Centralized and Cooperative Transmission of Secure Multiple Unicasts using Network Coding

We introduce a method for securely delivering a set of messages to a group of clients over a broadcast erasure channel where each client is interested in a distinct message. Each client is able to obtain its own message but not the others'. In the proposed method the messages are combined together using a special variant of random linear network coding. Each client is provided with a private set of decoding coefficients to decode its own message. Our method provides security for the transmission sessions against computational brute-force attacks and also weakly security in information theoretic sense. As the broadcast channel is assumed to be erroneous, the missing coded packets should be recovered in some way. We consider two different scenarios. In the first scenario the missing packets are retransmitted by the base station (centralized). In the second scenario the clients cooperate with each other by exchanging packets (decentralized). In both scenarios, network coding techniques are exploited to increase the total throughput. For the case of centralized retransmissions we provide an analytical approximation for the throughput performance of instantly decodable network coded (IDNC) retransmissions as well as numerical experiments. For the decentralized scenario, we propose a new IDNC based retransmission method where its performance is evaluated via simulations and analytical approximation. Application of this method is not limited to our special problem and can be generalized to a new class of problems introduced in this paper as the cooperative index coding problem

UV-isomerisation in nematic elastomers as a route to photo-mechanical transducer

The macroscopic shape of liquid crystalline elastomers strongly depends on the order parameter of the mesogenic groups. This order can be manipulated if photoisomerisable groups, e.g. containing N=N bonds, are introduced into the material. We have explored the large photo-mechanical response of such an azobenzene-containing nematic elastomer at different temperatures, using force and optical birefringence measurements, and focusing on fundamental aspects of population dynamics and the related speed and repeatability of the response. The characteristic time of ``on'' and ``off'' regimes strongly depends on temperature, but is generally found to be very long. We were able to verify that the macroscopic relaxation of the elastomer is determined by the nematic order dynamics and not, for instance, by the polymer network relaxation.Comment: Latex (EPJE class) 12 figure

The Impact of Selected Gene Mutations to the Activation and DNA Binding Activity of the Transcription Factor VraR

VraSR two-component system in S. aureus senses bacterial cell wall damage caused by antibiotics (vancomycin or -lactams) and regulates a set of genes. We have undertaken the study of the role of certain amino acids in the function of VraR, which undergo mutation in clinical S. aureus resistant strains and analyzed their effects on the VraSR signal transduction mechanism. In particular, we have looked at the role of Met-13 in VraR phosphorylation induced activation. M13A failure to be activated by phosphorylation resulted in a weaker binding to DNA. Moreover, we investigated VraR E59D, S164P, A113V, and S164A binding activity to the PvraSR by DNase-I footprinting. Finally, in vivo studies on the effects of substitutions of the above residues on the mRNA level of a number of VraSR regulon genes (fmtA, pbp2, and sgtb) provide insights as to how substitutions at certain positions may be beneficial to the S. aureus resistance to antibiotics