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Many bacterial histidine kinases work in two-component systems that combine into larger multi-kinase networks. NahK is one of the kinases in the GacS Multi-Kinase Network (MKN), which is the MKN that controls biofilm regulation in the opportunistic pathogenPseudomonas aeruginosa. This network has also been associated with regulating many virulence factorsP. aeruginosasecretes to cause disease. However, the individual role of each kinase is unknown. In this study, we identify NahK as a novel regulator of the phenazine pyocyanin (PYO). Deletion ofnahKleads to a fourfold increase in PYO production, almost exclusively through upregulation of phenazine operon two (phz2). We determined that this upregulation is due to mis-regulation of allP. aeruginosaquorum-sensing (QS) systems, with a large upregulation of thePseudomonasquinolone signal system and a decrease in production of the acyl-homoserine lactone-producing system,las. In addition, we see differences in expression of quorum-sensing inhibitor proteins that align with these changes. Together, these data contribute to understanding how the GacS MKN modulates QS and virulence and suggest a mechanism for cell density-independent regulation of quorum sensing.

Pseudomonas aeruginosais a Gram-negative bacterium that establishes biofilms as part of its pathogenicity.P. aeruginosainfections are associated with nosocomial infections. As the prevalence of multi-drug-resistantP. aeruginosaincreases, it is essential to understand underlying virulence molecular mechanisms. Histidine kinase NahK is one of several kinases inP. aeruginosaimplicated in biofilm formation and dispersal. Previous work has shown that the nitric oxide sensor, NosP, triggers biofilm dispersal by inhibiting NahK. The data presented here demonstrate that NahK plays additional important roles in theP. aeruginosalifestyle, including regulating bacterial communication mechanisms such as quorum sensing. These effects have larger implications in infection as they affect toxin production and virulence.

 

Developing a materials perspective of how to control the degradation and negative impact of complex metal oxides requires an integrated understanding of how these nanomaterials transform in the environment and interact with biological systems. Doping with aluminum is known to stabilize oxide materials, but has not been assessed cohesively from synthesis to environmental fate and biological impact. In the present study, the influence of aluminum doping on metal ion release from transition metal oxides was investigated by comparing aqueous transformations of lithium nickel cobalt aluminum oxide (LiNi0.82Co0.15Al0.03O2; NCA) and lithium nickel cobalt oxide (LiNi0.80Co0.20O2; NC) nanoparticles and by calculating the energetics of metal release using a density functional theory (DFT) and thermodynamics method. Two model environmental organisms were used to assess biological impact, and metal ion release was compared for NCA and NC nanoparticles incubated in their respective growth media: moderately hard reconstituted water (MHRW) for the freshwater invertebrate Daphnia magna (D. magna) and minimal growth medium for the Gram-negative bacterium Shewanella oneidensis MR-1 (S. oneidensis). The amount of metal ions released was reduced for NCA compared to NC in MHRW, which correlated to changes in the modeled energetics of release upon Al substitution in the lattice. In minimal medium, metal ion release was approximately an order of magnitude higher compared to MHRW and was similar to the stoichiometry of the bulk nanoparticles for both NCA and NC. Interpretation of the release profiles and modeling indicated that the increase in total metal ion release and the reduced influence of Al doping arises from lactate complexation of metal ions in solution. The relative biological impacts of NC and NCA exposure for both S. oneidensis and D. magna were consistent with the metal release trends observed for minimal medium and MHRW, respectively. Together, these results demonstrate how a combined experimental and computational approach provides valuable insight into the aqueous transformations and biological impacts of complex metal oxide nanoparticles. 

The use of engineered nanomaterials, defined as those smaller than 100 nm, in the health, energy, agricultural, and environmental sectors is expanding rapidly. As such, human and environmental exposure to these materials is increasing every day. For example, metal-based nanomaterials, such as nanosilver, have become ubiquitous in antibacterial applications ranging from socks and baby bottles to healthcare materials, such as oral fillings. Engineered nanomaterials are also used as antibacterial agents and adjuvants to improve antibiotic delivery or efficacy. However, even nanomaterials that were not designed to be antimicrobial can possess potent bactericidal activity. Alarmingly, there are clear connections between nanomaterial exposure, metal resistance, and antibiotic resistance and it is crucial that we dramatically improve our understanding of both the toxicity of these materials and their ability to permanently change the organisms that they encounter. Emerging research indicates that microbes are capable of adapting to nanomaterial toxicity, often with the same generalizable mechanisms used to overcome antibiotic toxicity. In this perspective, we highlight existing knowledge about microbial response to engineered nanomaterials and the key outstanding questions that must be addressed. 

Video games and immersive, narrative experiences are often called upon to help students understand difficult scientific concepts, such as sense of scale. However, the development of educational video games requires expertise and, frequently, a sizable budget. Here, we report on the use of an interactive text-style video game, NanoAdventure, to communicate about sense of scale and nanotechnology to the public. NanoAdventure was developed on an open-source, free-to-use platform with simple coding and enhanced with free or low-cost assets. NanoAdventure was launched in three languages (English, Spanish, Chinese) and compared to textbook-style and blog-style control texts in a randomized study. Participants answered questions on their knowledge of nanotechnology and their attitudes toward nanotechnology before and after reading one randomly assigned text (textbook, blog, or NanoAdventure game). Our results demonstrate that interactive fiction is effective in communicating about sense of scale and nanotechnology as well as the relevance of nanotechnology to a general public. NanoAdventure was found to be the most “fun” and easy to read of all text styles by participants in a randomized trial. Here, we make the case for interactive “Choose Your Own Adventure” style games as another effective tool among educational game models for chemistry and science communication. 

Engineered nanoparticles are incorporated into numerous emerging technologies because of their unique physical and chemical properties. Many of these properties facilitate novel interactions, including both intentional and accidental effects on biological systems. Silver-containing particles are widely used as antimicrobial agents and recent evidence indicates that bacteria rapidly become resistant to these nanoparticles. Much less studied is the chronic exposure of bacteria to particles that were not designed to interact with microorganisms. For example, previous work has demonstrated that the lithium intercalated battery cathode nanosheet, nickel manganese cobalt oxide (NMC), is cytotoxic and causes a significant delay in growth of Shewanella oneidensis MR-1 upon acute exposure. Here, we report that S. oneidensis MR-1 rapidly adapts to chronic NMC exposure and is subsequently able to survive in much higher concentrations of these particles, providing the first evidence of permanent bacterial resistance following exposure to nanoparticles that were not intended as antibacterial agents. We also found that when NMC-adapted bacteria were subjected to only the metal ions released from this material, their specific growth rates were higher than when exposed to the nanoparticle. As such, we provide here the first demonstration of bacterial resistance to complex metal oxide nanoparticles with an adaptation mechanism that cannot be fully explained by multi-metal adaptation. Importantly, this adaptation persists even after the organism has been grown in pristine media for multiple generations, indicating that S. oneidensis MR-1 has developed permanent resistance to NMC. 

Lithium nickel manganese cobalt oxide (Li x Ni y Mn z Co 1−y−z O 2 , 0 < x , y , z < 1, also known as NMC) is a class of cathode materials used in lithium ion batteries. Despite the increasing use of NMC in nanoparticle form for next-generation energy storage applications, the potential environmental impact of released nanoscale NMC is not well characterized. Previously, we showed that the released nickel and cobalt ions from nanoscale Li 1/3 Ni 1/3 Mn 1/3 Co 1/3 O 2 were largely responsible for impacting the growth and survival of the Gram-negative bacterium Shewanella oneidensis MR-1 (M. N. Hang et al. , Chem. Mater. , 2016, 28 , 1092). Here, we show the first steps toward material redesign of NMC to mitigate its biological impact and to determine how the chemical composition of NMC can significantly alter the biological impact on S. oneidensis . We first synthesized NMC with various stoichiometries, with an aim to reduce the Ni and Co content: Li 0.68 Ni 0.31 Mn 0.39 Co 0.30 O 2 , Li 0.61 Ni 0.23 Mn 0.55 Co 0.22 O 2 , and Li 0.52 Ni 0.14 Mn 0.72 Co 0.14 O 2 . Then, S. oneidensis were exposed to 5 mg L −1 of these NMC formulations, and the impact on bacterial oxygen consumption was analyzed. Measurements of the NMC composition, by X-ray photoelectron spectroscopy, and composition of the nanoparticle suspension aqueous phase, by inductively coupled plasma-optical emission spectroscopy, showed the release of Li, Ni, Mn, and Co ions. Bacterial inhibition due to redesigned NMC exposure can be ascribed largely to the impact of ionic metal species released from the NMC, most notably Ni and Co. Tuning the NMC stoichiometry to have increased Mn at the expense of Ni and Co showed lowered, but not completely mitigated, biological impact. This study reveals that the chemical composition of NMC nanomaterials is an important parameter to consider in sustainable material design and usage.