Engineered polyethersulfone membrane for well-dispersed silver nanoparticle impregnation at high loading: high water permeate flux and biofouling prevention.

We present a new method for impregnation of silver nanoparticles (Ag NPs) at high loading on polyethersulfone (PES) membrane's external surface, simultaneously retaining native membrane's porosity - to achieve a high water permeate flux without biofouling. This was possible by PES membrane's surface modification with acrylic acid (AA), finally leading to AA-Ag-PES membrane. AA-Ag-PES had a high (9.04%) Ag-NP loading selectively on membrane surface, as discrete, smaller (mean size: 20 nm) nanoparticles (NPs). In nonfunctionalized Ag-PES, aggregated (mean size: 70 nm) NPs, with lower Ag loading (0.73 wt.%) was obtained, with NP being present both on membrane surface and inside pores. Consequently, AA-Ag-PES could maintain similar water permeability and porosity (10,153.05 Lm-2 h-1bar-1 and 69.98%, respectively), as in native PES (11,368.74 Lm-2 h-1bar-1 and 68.86%, respectively); whereas both parameters dropped significantly for Ag-PES (4,869.66 Lm-2 h-1bar-1 and 49.02%, respectively). AA-Ag-PES also showed least flux reduction (7.7%) due to its anti-biofouling property and high flux recovery after usage and cleaning, compared to native PES and Ag-PES membrane's much higher flux reduction (54.29% and 36.7%, respectively). Hence, discrete NP impregnation, avoiding pore blockage, is key for achieving high water flux and anti-biofouling properties (in AA-Ag-PES), compared to non-functionalized Ag-PES, due to aggregated Ag-NPs inside its pores.

Uncoupling of invasive bacterial mucosal immunogenicity from pathogenicity.

There is the notion that infection with a virulent intestinal pathogen induces generally stronger mucosal adaptive immunity than the exposure to an avirulent strain. Whether the associated mucosal inflammation is important or redundant for effective induction of immunity is, however, still unclear. Here we use a model of auxotrophic Salmonella infection in germ-free mice to show that live bacterial virulence factor-driven immunogenicity can be uncoupled from inflammatory pathogenicity. Although live auxotrophic Salmonella no longer causes inflammation, its mucosal virulence factors remain the main drivers of protective mucosal immunity; virulence factor-deficient, like killed, bacteria show reduced efficacy. Assessing the involvement of innate pathogen sensing mechanisms, we show MYD88/TRIF, Caspase-1/Caspase-11 inflammasome, and NOD1/NOD2 nodosome signaling to be individually redundant. In colonized animals we show that microbiota metabolite cross-feeding may recover intestinal luminal colonization but not pathogenicity. Consequent immunoglobulin A immunity and microbial niche competition synergistically protect against Salmonella wild-type infection.

Mechanistic insight into ROS and neutral lipid alteration induced toxicity in the human model with fins (Danio rerio) by industrially synthesized titanium dioxide nanoparticles.

The toxicological impact of TiO2 nanoparticles on the environment and human health has been extensively studied in the last few decades, but the mechanistic details were unknown. In this study, we evaluated the impact of industrially prepared TiO2 nanoparticles on the biological system using zebrafish embryo as an in vivo model. The industrial synthesis of TiO2 nanoparticles was mimicked on the lab scale using the high energy ball milling (HEBM) method by milling bulk TiO2 particles for 5 h, 10 h, and 15 h in an ambient environment. The physiochemical properties were characterized by standard methods like field emission scanning electron microscopy (FESEM), dynamic light scattering (DLS), X-ray diffraction (XRD) and UV-Visible spectroscopy. In vivo cytotoxicity was assessed on zebrafish embryos by the evaluation of their mortality rate and hatching rate. Experimental and computational analysis of reactive oxygen species (ROS) induction, apoptosis, and neutral lipid alteration was done to study the effects on the cellular level of zebrafish larvae. The analysis depicted the change in size and surface charge of TiO2 nanoparticles with respect to the increase in milling time. In silico investigations revealed the significant role of ROS quenching and altered neutral lipid accumulation functionalised by the molecular interaction of respective metabolic proteins in the cytotoxicity of TiO2 nanoparticles with zebrafish embryos. The results reveal the hidden effect of industrially synthesized TiO2 nanoparticle exposure on the alteration of lipid accumulation and ROS in developing zebrafish embryos. Moreover, the assessment provided a detailed mechanistic analysis of in vivo cytotoxicity at the molecular level.

Biological and regulatory roles of acid-induced small RNA RyeC in Salmonella Typhimurium.

Salmonella Typhimurium is an enteric pathogen that has evolved masterful strategies to enable survival under stress conditions both within and outside a host. The acid tolerance response (ATR) is one such mechanism that enhances the viability of acid adapted bacteria to lethal pH levels. While numerous studies exist on the protein coding components of this response, there is very little data on the roles of small RNAs (sRNAs). These non-coding RNA molecules have recently been shown to play roles as regulators of bacterial stress response and virulence pathways. They function through complementary base pairing interactions with target mRNAs and affect their translation and/or stability. There are also a few that directly bind to proteins by mimicking their respective targets. Here, we identify several sRNAs expressed during the ATR of S. Typhimurium and characterize one highly induced candidate, RyeC. Further, we identify ptsI as a trans-encoded target that is directly regulated by this sRNA. From a functional perspective, over-expression of RyeC in Salmonella produced a general attenuation of several in vitro phenotypes including acid survival, motility, adhesion and invasion of epithelial cell lines as well as replication within macrophages. Together, this study highlights the diverse roles played by sRNAs in acid tolerance and virulence of S. Typhimurium.

The expanding targetome of small RNAs in Salmonella Typhimurium.

The enterobacterial pathogen Salmonella has long served as a model for bacterial pathogenesis, stress response, gene expression and regulation with extensive investigation involving protein function. With the advent of high-throughput sequencing technologies and their applications in genomics and transcriptomics, a wealth of data particularly with respect to small RNAs (sRNAs) is being generated. These molecules serve as regulators of major stress response and virulence networks in diverse species including Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) and have received a lot of focus in the last decade. sRNAs are major post-transcriptional regulators that either repress or activate their targets, the former comprising the bulk of known interactions. In this review, we update and summarize validated sRNA-target regulatory information in S. Typhimurium. It covers the largest group of sRNAs in this pathogen, namely trans-encoded antisense RNAs which have received a lot of focus over the past decade. Additionally, we explore the role of Hfq as a global RNA-binding protein and highlight current techniques developed to identify sRNA targets.