Ambient particulate matter exposure induces ferroptosis in hippocampal cells through the GSK3B/Nrf2/GPX4 pathway.

Epidemiological studies have established a robust correlation between exposure to ambient particulate matter (PM) and various neurological disorders, with dysregulation of intracellular redox processes and cell death being key mechanisms involved. Ferroptosis, a cell death form characterized by iron-dependent lipid peroxidation and disruption of antioxidant defenses, may be involved in the neurotoxic effects of PM exposure. However, the relationship between PM-induced neurotoxicity and ferroptosis in nerve cells remains to be elucidated. In this study, we utilized a rat model (exposed to PM at a dose of 10 mg/kg body weight per day for 4 weeks) and an HT-22 cell model (exposed to PM at concentrations of 50, 100, and 200 µg/mL for 24 h) to investigate the potential induction of ferroptosis by PM exposure. Furthermore, RNA sequencing analysis was employed to identify hub genes that potentially contribute to the process of ferroptosis, which was subsequently validated through in vivo and in vitro experiments. The results revealed that PM exposure increased MDA content and Fe2+ levels, and decreased SOD activity and GSH/GSSG ratio in rat hippocampal and HT-22 cells. Through RNA sequencing analysis, bioinformatics analysis, and RT-qPCR experiments, we identified GSK3B as a possible hub gene involved in ferroptosis. Subsequent investigations demonstrated that PM exposure increased GSK3B levels and decreased Nrf2, and GPX4 levels in vivo and in vitro. Furthermore, treatment with LY2090314, a specific inhibitor of GSK3B, was found to mitigate the PM-induced elevation of MDA and ROS and restore SOD activity and GSH/GSSG ratio. The LY2090314 treatment promoted the upregulation of Nrf2 and GPX4 and facilitated the nuclear translocation of Nrf2 in HT-22 cells. Moreover, treatment with LY2090314 resulted in the upregulation of Nrf2 and GPX4, along with the facilitation of nuclear translocation of Nrf2. This study suggested that PM-induced ferroptosis in hippocampal cells may be via the GSK3B/Nrf2/GPX4 pathway.

TREM2 mitigates NLRP3-mediated neuroinflammation through the NF-κB and PI3k/Akt signaling pathways in juvenile rats exposed to ambient particulate matter.

Ambient particulate matter (PM) is a global public and environmental problem. PM is closely associated with several neurological disorders that typically involve neuroinflammation. There have been few studies on the effect of PM on neuroinflammation to date. In this study, we used a juvenile rat model (PM exposure was conducted at a dose of 10 mg/kg body weight per day for 4 weeks) and a BV-2 cell model (PM exposure was conducted at concentrations of 50, 100, 150, and 200 µg/ml for 24 h) to investigate PM-induced neuroinflammation mediated by NLRP3 inflammasome activation and the role of TREM2 in this process. Our findings revealed that PM exposure reduced TREM2 protein and mRNA levels in the rat hippocampus and BV-2 cells. TREM2 overexpression attenuated PM-induced spatial learning and memory deficits in rats. Moreover, we observed that TREM2 overexpression in vivo and in vitro effectively mitigated the increase in NLRP3 and pro-Caspase1 protein expression, as well as the secretion of IL-1ß and IL-18. Exposure to PM increased the expression of NF-κB and decreased the phosphorylation of PI3k/Akt in vivo and in vitro, and this process was effectively reversed by overexpressing TREM2. Our results indicated that PM exposure could reduce TREM2 expression and induce NLRP3 inflammasome-mediated neuroinflammation and that TREM2 could mitigate NLRP3 inflammasome-mediated neuroinflammation by regulating the NF-κB and PI3k/Akt signaling pathways. These findings shed light on PM-induced neuroinflammation mechanisms and potential intervention targets.

Impact of amino acid substitutions on the behavior of a photoactivatable near infrared fluorescent protein PAiRFP1.

A photoactivatable near-infrared fluorescent protein (NIR-FP) PAiRFP1 has been developed by 15 amino acid substitutions in its nonfluorescent template Agp2. In our previous communication, we investigated the role of three amino acids in PHY domain distal from BV molecule. The impact of the twelve amino acids in GAF domain, especially five residues near BV-binding pocket is unclear. In this paper, PCR based reverse mutagenesis, spectroscopic methods, molecular modelling and simulations have been employed to explore the roles of these substitutions during the molecular evolution of PAiRFP1. It was found that the residue L163 is important for protein folding in PAiRFP1. The residues F244 and C280 exerted remarkable effects on molar extinction coefficient, NIR fluorescence quantum yield, molecular brightness, fluorescence fold, and dark recovery rate. The residues F244 and V276 modulate the maximum absorption and emission peak position. The reverse mutant L168M exhibited a higher fluorescence fold than PAiRFP1. Additionally, the reverse mutants V203A, V294E, S218G and D127G possessed better spectral properties than PAiRFP1. This study is important for the rational design of a better BphP-based photoactivatable NIR-FPs.

Application of two-dimensional correlation fluorescence spectroscopy to detect the presence of trace amount of substances.

Although fluorescence spectroscopy is a highly sensitive method, it is still rather difficult to identify a minor fluorescent component whose fluorescent peak is overlapped and masked by a dominant fluorescent component in a sample solution. Herein, we describe a two-dimensional correlation spectroscopy (2D-COS) approach based on the Kasha's rule to solve the above common problem. We initially suppose that a sample solution contains the major component only, and the spectral behavior of the major component obeys the Kasha's rule. Then, the shapes of emission spectra obtained under excitation lights of different wavelengths remain invariant. Under this condition, the introduction of a minor fluorescent component can be reflected by the changes on the shapes of emission peaks in the series of emission spectra. Moreover, subtle changes, which are difficult to be found in the original spectra, can be clearly visualized as cross peaks in 2D asynchronous spectrum constructed using a series of emission spectra. In addition, we demonstrate that the intensities of cross peaks can be enhanced by changing the sequence of the series of emission spectra. We utilize the approach on an aqueous solution containing eosin Y and a trace amount of bromocresol green. The presence of bromocresol green with the concentration as low as 400 nM can be revealed via the cross peaks in the resultant 2D asynchronous spectra. In a preliminary study, we suggest that 2D disrelation spectrum might provide an alternative chance to reveal the presence of small amount bromocresol green.

Effects of reverse genetic mutations on the spectral and photochemical behavior of a photoactivatable fluorescent protein PAiRFP1.

Bacteriophytochrome photoreceptors (BphPs) containing biliverdin (BV) have great potential for the development of genetically engineered near-infrared fluorescent proteins (NIR FPs). We investigated a photoactivatable fluorescent protein PAiRFP1, was engineered through directed molecular evolution. The coexistence of both red light absorbing (Pr) and far-red light absorbing (Pfr) states in dark is essential for the photoactivation of PAiRFP1. The PCR based site-directed reverse mutagenesis, spectroscopic measurements and molecular dynamics (MD) simulations were performed on three targeted sites V386A, V480A and Y498H in PHY domain to explore their potential effects during molecular evolution of PAiRFP1. We found that these substitutions did not affect the coexistence of Pr and Pfr states but led to slight changes in the photophysical parameters. The covalent docking of biliverdin (cis and trans form) with PAiRFP1 was followed by several 100 ns MD simulations to provide some theoretical explanations for the coexistence of Pr and pfr states. The results suggested that experimentally observed coexistence of Pr and Pfr states in both PAiRFP1 and mutants were resulted from the improved stability of Pr state. The use of experimental and computational work provided useful understanding of Pr and Pfr states and the effects of these mutations on the photophysical properties of PAiRFP1.

Nitrogen and sulfur-codoped porous carbon derived from a BSA/ionic liquid polymer complex: multifunctional electrode materials for water splitting and supercapacitors.

Bovine serum albumin (BSA) was complexed with a hydrophobic ionic liquid polymer (PIL) via electrostatic interaction to fabricate a carbon precursor. Then, a novel nitrogen (N) and sulfur (S) codoped micro-/mesoporous carbon (NSPC) was obtained via direct carbonization of the interpolyelectrolyte BSA@PIL complex. The newly developed NSPC materials exhibited excellent HER/OER electrocatalytic activity and stability, as well as outstanding capacitance performance. Remarkably, NSPC pyrolyzed at 1000 degrees (NSPC-1000) presented an overpotential as low as 172 mV vs. RHE (without iR correction) to achieve a current density of 10 mA cm-2 and a Tafel slope of 44.3 mV dec-1 in 0.5 M H2SO4 for HER, as well as a low overpotential of 460 mV vs. RHE in 0.1 M KOH for OER. Furthermore, NSPC-1000 offers a specific capacitance as high as 495 F g-1 at a current density of 0.1 A g-1. Such excellent performance of NSPC in electrocatalytic water splitting and supercapacitors originates from the synergistic effects of its N/S-codoping and micro-/mesoporous hierarchical architecture. Our facile protocol through combining biomacromolecules and synthetic polymers offers a new strategy in the development of effective, readily scalable and metal-free heteroatom-doped carbon materials for energy-related applications.

Electrochemical nitrogen reduction to ammonia at ambient conditions on nitrogen and phosphorus co-doped porous carbon.

Reduction of nitrogen (N2) under ambient conditions and understanding the mechanism have been hugely challenging problems for decades. Herein, we for the first time report that N,P co-doped hierarchical porous carbon (NPC) can serve as an electrocatalyst for the nitrogen reduction reaction (NRR) in an acid aqueous solution under ambient conditions. The faradaic efficiency (FE) and yield of production of NH3 on the NPC electrode reached as high as 4.2% and 0.97 µg h-1 mg-1cat., respectively. Furthermore, the electrocatalytic NRR mechanism on the NPC electrode was undoubtedly confirmed by electrochemical in situ Fourier transform infrared spectroscopy, and follows an associative pathway. These results are predicted to offer a new platform in the rational design and synthesis of highly efficient electrocatalysts for the NRR under ambient conditions.