Antimicrobial efficacy, which is central to many aspects of medicine, is being rapidly eroded by bacterial resistance. Since new resistance can be induced by antimicrobial action, highly lethal agents that rapidly reduce bacterial burden during infection should help restrict the emergence of resistance. To improve lethal activity, recent work has focused on toxic reactive oxygen species (ROS) as part of the bactericidal activity of diverse antimicrobials. We report that when Escherichia coli was subjected to antimicrobial stress and the stressor was subsequently removed, both ROS accumulation and cell death continued to occur. Blocking ROS accumulation by exogenous mitigating agents slowed or inhibited poststressor death. Similar results were obtained with a temperature-sensitive mutational inhibition of DNA replication. Thus, bacteria exposed to lethal stressors may not die during treatment, as has long been thought; instead, death can occur after plating on drug-free agar due to poststress ROS-mediated toxicity. Examples are described in which (i) primary stress-mediated damage was insufficient to kill bacteria due to repair; (ii) ROS overcame repair (i.e., protection from anti-ROS agents was reduced by repair deficiencies); and (iii) killing was reduced by anti-oxidative stress genes acting before stress exposure. Enzymatic suppression of poststress ROS-mediated lethality by exogenous catalase supports a causal rather than a coincidental role for ROS in stress-mediated lethality, thereby countering challenges to ROS involvement in antimicrobial killing. We conclude that for a variety of stressors, lethal action derives, at least in part, from stimulation of a self-amplifying accumulation of ROS that overwhelms the repair of primary damage.reactive oxygen species | poststress cellular response | antimicrobial | antioxidant | damage repair D iscovering ways to manage antimicrobial resistance is among the most important medical challenges of our time (1). Since many antimicrobials can stimulate the production of resistant mutants, often via the SOS response (2-8), one way to limit the emergence of new resistance is to more rapidly and extensively reduce pathogen populations during infection. Toward that end, we and others have been studying how antimicrobials and other lethal stressors kill bacteria. Recent work has drawn attention to the contribution of stress-stimulated accumulation of toxic reactive oxygen species (ROS) (9-21). Finding ways to stimulate ROS-mediated killing could in principle enhance the efficacy of a broad range of antimicrobials. However, an ROS contribution to antimicrobial killing became controversial when the original observation (9) was challenged (22-24). Subsequent work countered many of the challenges (13)(14)(15)25) and extended the phenomenon to thymineless death (26), phage infection (27), the type VI secretion system (27), and overexpression of a MalE-LacZ fusion (28). Moreover, nitric oxide and hydrogen sulfide interfere with antimicrobial killing by suppressing ROS generation/ac...

No abstract

Protein lysine succinylation, an emerging protein post-translational modification widespread among eukaryotic and prokaryotic cells, represents an important regulator of cellular processes. However, the extent and function of lysine succinylation in Mycobacterium tuberculosis, especially extensively drug-resistant strain, remain elusive. Combining protein/peptide prefractionation, immunoaffinity enrichment, and LC-MS/MS analysis, a total of 686 succinylated proteins and 1739 succinylation sites of M. tuberculosis were identified, representing the first global profiling of M. tuberculosis lysine succinylation. The identified succinylated proteins are involved in a variety of cellular functions such as metabolic processes, transcription, translation, and stress responses and exhibit different subcellular localization via GO, protein interaction network, and other bioinformatic analysis. Notably, proteins involved in protein biosynthesis and carbon metabolism are preferred targets of lysine succinylation. Moreover, two prevalent sequence patterns: EK(suc) and K*****K(suc), can be found around the succinylation sites. There are 109 lysine-succinylated homologues in E. coli, suggesting highly conserved succinylated proteins. Succinylation was found to occur at the active sites predicted by Prosite signature including Rv0946c, indicating that lysine succinylation may affect their activities. There is extensive overlapping between acetylation sites and succinylation sites in M. tuberculosis. Many M. tuberculosis metabolic enzymes and antibiotic resistance proteins were succinylated. This study provides a basis for further characterization of the pathophysiological role of lysine succinylation in M. tuberculosis.

No abstract

Massive connectivity and low latency are two important challenges for the Internet of Things (IoT) to achieve the Quality of Service (QoS) provisions required by the numerous devices it is designed to service. Motivated by these challenges, in the paper we introduce a new millimeter-wave non-orthogonal multiple access (mmWave-NOMA) transmission scheme designed for cellular machine-to-machine (M2M) communication systems for IoT applications. It consists of one base station (BS) and numerous multiple machine type communication (MTC) devices operating in a cellular communication environment. We consider its down-link performance and assume that multiple MTC devices share the same communication resources offered by the proposed mmWave-NOMA transmission scheme, which can support massive connectivity. For this system, a novel MTC pairing scheme is introduced the design of which is based upon the distance between the BS and the MTC devices aiming at reducing the system overall overhead for massive connectivity and latency. In particular, we consider three different MTC device pairing schemes, namely i) the random near and the random far MTC devices (RNRF); ii) the nearest near and the nearest far MTC devices (NNNF); and iii) the nearest near and the farthest far MTC device (NNFF). For all three pairing schemes, their performance is analyzed by deriving closed-form expressions of the outage probability and the sum rate. Furthermore, performance comparison studies of the three MTC device pairing schemes have been carried out. The validity of the analytical approach has been verified by means of extensive computer simulations. The obtained performance evaluation results have demonstrated that the proposed cellular M2M communication system employing the mmWave-NOMA transmission scheme improves outage probability as compared to equivalent systems using mmWave with Orthogonal Multiple Access (OMA) schemes.Index Terms-Internet of Things (IoT), millimeter-wave non-orthogonal multiple access (mmWave-NOMA), machine-tomachine (M2M), MTC device pairing schemes, outage probability.

Human endogenous retrovirus (HERV) accounts for ∼8% of the human genome. Recent studies have reported that multiple HERV genes and long terminal repeats (LTRs) are involved in human tumorigenesis. Here we demonstrated that HERV-W env (syncytin-1) was overexpressed in 75.6% (62/82) of urothelial cell carcinoma (UCC) tissues of the bladder compared with only 6.1% (5/82) of matched tumor-adjacent tissues (P<0.001). Syncytin-1 overexpression increased proliferation and viability of immortalized human uroepithelial cells. Colony-formation experiments and in-vivo tumor xenografts suggested that syncytin-1 overexpression had oncogenic potential. Syncytin-1 3'-LTR mutations (142T>C and 277A>G) were present in 87.8% (72/82) of UCC tissues. Normal 3'-LTR was found in 12.2% (10/82) of UCC tissues compared with 95.1% (78/82) of matched tumor-adjacent tissues (P<0.001). Interestingly, 3'-LTR mutations were significantly associated with syncytin-1 overexpression. Luciferase assay and expression analysis revealed that 3'-LTR mutations, especially the 142T>C mutation, enhanced the syncytin-1 promoter activity and expression. In-silico analysis, electrophoretic mobility shift assays and chromatin immunoprecipitation assays demonstrated the binding of c-Myb to 3'-LTRs when the mutations occurred. This alternative interaction was found to be dependent on 142T>C mutation. C-Myb activated syncytin-1 promoter activity and expression by binding to mutant 3'-LTRs. Taken together, these data indicate that syncytin-1 overexpression may be an indicator of UCC risk. The 3'-LTR mutations may upregulate syncytin-1 expression, enabling it to participate in UCC tumorigenesis and development by interacting with c-Myb.

The success of Mycobacterium tuberculosis (M. tuberculosis) as a pathogen is largely contributes to its ability to manipulate the host immune responses. The genome of M. tuberculosis encodes multiple immune-modulatory proteins, including several members of the multi-genic PE_PPE family. Despite of intense research, the roles of PE_PGRS proteins in mycobacterial pathogenesis remain elusive. The function of M. tuberculosis PE_PGRS41, characterized by an extended and unique C-terminal domain, was studied. Expression of PE_PGRS41 in Mycobacterium smegmatis, a non-pathogenic species intrinsically deficient of PE_PGRS, severely impaired the resistance of the recombinant to multiple stresses via altering the cell wall integrity. Macrophages infected by M. smegmatis harboring PE_PGRS41 decreased the production of TNF-α, IL-1β and IL-6. In addition, PE_PGRS41 boosted the survival of M. smegmatis within macrophage accompanied with enhanced cytotoxic cell death through inhibiting the cell apoptosis and autophagy. Taken together, these results implicate that PE_PGRS41 is a virulence factor of M. tuberculosis and sufficient to confer pathogenic properties to M. smegmatis.

No abstract