Long Noncoding RNA Metastasis-Associated Lung Adenocarcinoma Transcript 1 Promotes HIV-1 Replication through Modulating microRNAs in Macrophages.

Macrophages can serve as a reservoir for human immunodeficiency-1 (HIV-1) virus in host cells, constituting a barrier to eradication, even in patients who are receiving antiretroviral therapy. Although many noncoding RNAs have been characterized as regulators in HIV-1/AIDS-induced immune response and pathogenesis, only a few long noncoding RNAs (lncRNAs) have demonstrated a close association with HIV-1 replication, and the molecular mechanisms remain unknown. In this study, we investigated how lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), related microRNAs, and key inflammatory genes alter HIV-1 replication in macrophages. Our data show that HIV-1 infection modulates the expression of miR-155 and miR-150-5p in a time-dependent manner, which is regulated by MALAT1. MALAT1 induced suppressor of cytokine signaling 1 (SOCS1) expression by sponging miR-150-5p in HIV-1-infected macrophages and stimulated inflammatory mediators triggering receptor expressed on myeloid cells/cold inducible RNA binding protein (TREM 1/CIRP) ligand/receptor. The RNA immunoprecipitation (RIP) assay validated the direct interaction within the MALAT1/miR-150-5p/SOCS1 axis. HIV-1 infection-mediated upregulation of MALAT1, SOCS1, and HIV-1 Gag was attenuated by SN50 (an NF-кB p50 inhibitor). MALAT1 antisense oligonucleotides (ASOs) suppressed HIV-1 p24 production and HIV-1 Gag gene expression and decreased expression of miR-155 and SOCS1, as well as the production of proinflammatory cytokines by HIV-1-infected macrophages. In conclusion, HIV-1 infection induces MALAT1, which attenuates miR-150-5p expression and increases SOCS1 expression, promoting HIV-1 replication and reactivation. These data provide new insights into how MALAT1 alters the macrophage microenvironment and subsequently promotes viral replication and suggest a potential role for targeting MALAT1 as a therapeutic approach to eliminate HIV-1 reservoirs. IMPORTANCE Viral reservoirs constitute an obstacle to curing HIV-1 diseases, despite antiretroviral therapy. Macrophages serve as viral reservoirs in HIV infection by promoting long-term replication and latency. Recent studies have shown that lncRNAs can modulate virus-host interactions, but the underlying mechanisms are not fully understood. In this study, we demonstrate how lncRNA MALAT1 contributes to HIV-1 replication through modulation of the miR-150/SOCS1 axis in human macrophages. Our findings have the potential to identify new therapies for eliminating HIV-1 reservoirs in immune cells.

Computational design of a new layered superconductor LaOTlF2.

A new layered compound LaOTlF2 is designed and investigated using first-principles calculations in this work. The parent compound is an insulator with an indirect band gap of 3.88 eV. Electron-doping of the parent compound makes the material metallic. In the meantime, several lattice vibrational modes couple strongly to the conduction band, leading to a large electron-phonon coupling constant and conventional superconductivity. The highest superconducting transition temperature Tc is predicted to be approximately 8.6 K with λ about 1.25 in the optimally doped LaO0.95F0.05TlF2, where λ is calculated using the Wannier interpolation technique.

Perspectives on the flexibility analysis for continuous pharmaceutical manufacturing processes.

Pharmaceutical continuous manufacturing, especially under the context of COVID-19 pandemic, is regarded as an emerging technology that can guarantee the adequate quality assurance and mitigate process risk while guaranteeing the desirable economic performance. Flexibility analysis is one approach to quantitively assess the capability of chemical process to guarantee feasible operation in face of variations on uncertain parameters. The aim of this paper is to provide the perspectives on the flexibility analysis for continuous pharmaceutical manufacturing processes. State-of-the-art and progress in the flexibility analysis for chemical processes including concept evolution, mathematical model formulations, solution strategies, and applications are systematically overviewed. Recent achievements on the flexibility/feasibility analysis of the downstream dosage form manufacturing process are also touched upon. Further challenges and developments in the field of flexibility analysis for novel continuous manufacturing processes of active pharmaceutical ingredients along with the integrated continuous manufacturing processes are identified.

UPR modulation of host immunity by Pseudomonas aeruginosa in cystic fibrosis.

Cystic fibrosis (CF) is a progressive multiorgan autosomal recessive disease with devastating impact on the lungs caused by derangements of the CF transmembrane conductance regulator (CFTR) gene. Morbidity and mortality are caused by the triad of impaired mucociliary clearance, microbial infections and chronic inflammation. Pseudomonas aeruginosa is the main respiratory pathogen in individuals with CF infecting most patients in later stages. Despite its recognized clinical impact, molecular mechanisms that underlie P. aeruginosa pathogenesis and the host response to P. aeruginosa infection remain incompletely understood. The nuclear hormone receptor peroxisome proliferator-activated receptor (PPAR) γ (PPARγ), has shown to be reduced in CF airways. In the present study, we sought to investigate the upstream mechanisms repressing PPARγ expression and its impact on airway epithelial host defense. Endoplasmic reticulum-stress (ER-stress) triggered unfolded protein response (UPR) activated by misfolded CFTR and P. aeruginosa infection contributed to attenuated expression of PPARγ. Specifically, the protein kinase RNA (PKR)-like ER kinase (PERK) signaling pathway led to the enhanced expression of the CCAAT-enhancer-binding-protein homologous protein (CHOP). CHOP induction led to the repression of PPARγ expression. Mechanistically, we showed that CHOP induction mediated PPARγ attenuation, impacted the innate immune function of normal and ∆F508 primary airway epithelial cells by reducing expression of antimicrobial peptide (AMP) and paraoxanse-2 (PON-2), as well as enhancing IL-8 expression. Furthermore, mitochondrial reactive oxygen species production (mt-ROS) and ER-stress positive feedforward loop also dysregulated mitochondrial bioenergetics. Additionally, our findings implicate that PPARγ agonist pioglitazone (PIO) has beneficial effect on the host at the multicellular level ranging from host defense to mitochondrial re-energization.

Peroxisome proliferator-activated receptor-γ agonists attenuate biofilm formation by Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a significant contributor to recalcitrant multidrug-resistant infections, especially in immunocompromised and hospitalized patients. The pathogenic profile of P. aeruginosa is related to its ability to secrete a variety of virulence factors and to promote biofilm formation. Quorum sensing (QS) is a mechanism wherein P. aeruginosa secretes small diffusible molecules, specifically acyl homo serine lactones, such as N-(3-oxo-dodecanoyl)-l-homoserine lactone (3O-C12-HSL), that promote biofilm formation and virulence via interbacterial communication. Strategies that strengthen the host's ability to inhibit bacterial virulence would enhance host defenses and improve the treatment of resistant infections. We have recently shown that peroxisome proliferator-activated receptor γ (PPARγ) agonists are potent immunostimulators that play a pivotal role in host response to virulent P. aeruginosa Here, we show that QS genes in P. aeruginosa (strain PAO1) and 3O-C12-HSL attenuate PPARγ expression in bronchial epithelial cells. PAO1 and 3O-C12-HSL induce barrier derangements in bronchial epithelial cells by lowering the expression of junctional proteins, such as zonula occludens-1, occludin, and claudin-4. Expression of these proteins was restored in cells that were treated with pioglitazone, a PPARγ agonist, before infection with PAO1 and 3O-C12-HSL. Barrier function and bacterial permeation studies that have been performed in primary human epithelial cells showed that PPARγ agonists are able to restore barrier integrity and function that are disrupted by PAO1 and 3O-C12-HSL. Mechanistically, we show that these effects are dependent on the induction of paraoxonase-2, a QS hydrolyzing enzyme, that mitigates the effects of QS molecules. Importantly, our data show that pioglitazone, a PPARγ agonist, significantly inhibits biofilm formation on epithelial cells by a mechanism that is mediated via paraoxonase-2. These findings elucidate a novel role for PPARγ in host defense against P. aeruginosa Strategies that activate PPARγ can provide a therapeutic complement for treatment of resistant P. aeruginosa infections.-Bedi, B., Maurice, N. M., Ciavatta, V. T., Lynn, K. S., Yuan, Z., Molina, S. A., Joo, M., Tyor, W. R., Goldberg, J. B., Koval, M., Hart, C. M., Sadikot, R. T. Peroxisome proliferator-activated receptor-γ agonists attenuate biofilm formation by Pseudomonas aeruginosa.

Peroxisome proliferator-activated receptor-γ enhances human pulmonary artery smooth muscle cell apoptosis through microRNA-21 and programmed cell death 4.

Pulmonary hypertension (PH) is a progressive disorder whose cellular pathogenesis involves enhanced smooth muscle cell (SMC) proliferation and resistance to apoptosis signals. Existing evidence demonstrates that the tumor suppressor programmed cell death 4 (PDCD4) affects patterns of cell growth and repair responses in the systemic vasculature following experimental injury. In the current study, the regulation PDCD4 and its functional effects on growth and apoptosis susceptibility in pulmonary artery smooth muscle cells were explored. We previously demonstrated that pharmacological activation of the nuclear transcription factor peroxisome proliferator-activated receptor-γ (PPARγ) attenuated hypoxia-induced proliferation of human pulmonary artery smooth muscle cells (HPASMCs) by inhibiting the expression and mitogenic functions of microRNA-21 (miR-21). In the current study, we hypothesize that PPARγ stimulates PDCD4 expression and HPASMC apoptosis by inhibiting miR-21. Our findings demonstrate that PDCD4 is reduced in the mouse lung upon exposure to chronic hypoxia (10% O2 for 3 wk) and in hypoxia-exposed HPASMCs (1% O2). HPASMC apoptosis was reduced by hypoxia, by miR-21 overexpression, or by siRNA-mediated PPARγ and PDCD4 depletion. Activation of PPARγ inhibited miR-21 expression and resultant proliferation, while restoring PDCD4 levels and apoptosis to baseline. Additionally, pharmacological activation of PPARγ with rosiglitazone enhanced PDCD4 protein expression and apoptosis in a dose-dependent manner as demonstrated by increased annexin V detection by flow cytometry. Collectively, these findings demonstrate that PPARγ confers growth-inhibitory signals in hypoxia-exposed HPASMCs through suppression of miR-21 and the accompanying derepression of PDCD4 that augments HPASMC susceptibility to undergo apoptosis.

Enhanced Clearance of Pseudomonas aeruginosa by Peroxisome Proliferator-Activated Receptor Gamma.

The pathogenic profile of Pseudomonas aeruginosa is related to its ability to secrete a variety of virulence factors. Quorum sensing (QS) is a mechanism wherein small diffusible molecules, specifically acyl-homoserine lactones, are produced by P. aeruginosa to promote virulence. We show here that macrophage clearance of P. aeruginosa (PAO1) is enhanced by activation of the nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARγ). Macrophages treated with a PPARγ agonist (pioglitazone) showed enhanced phagocytosis and bacterial killing of PAO1. It is known that PAO1 QS molecules are inactivated by PON-2. QS molecules are also known to inhibit activation of PPARγ by competitively binding PPARγ receptors. In accord with this observation, we found that infection of macrophages with PAO1 inhibited expression of PPARγ and PON-2. Mechanistically, we show that PPARγ induces macrophage paraoxonase 2 (PON-2), an enzyme that degrades QS molecules produced by P. aeruginosa Gene silencing studies confirmed that enhanced clearance of PAO1 in macrophages by PPARγ is PON-2 dependent. Further, we show that PPARγ agonists also enhance clearance of P. aeruginosa from lungs of mice infected with PAO1. Together, these data demonstrate that P. aeruginosa impairs the ability of host cells to mount an immune response by inhibiting PPARγ through secretion of QS molecules. These studies define a novel mechanism by which PPARγ contributes to the host immunoprotective effects during bacterial infection and suggest a role for PPARγ immunotherapy for P. aeruginosa infections.

TREM-1-accentuated lung injury via miR-155 is inhibited by LP17 nanomedicine.

Triggering receptors expressed on myeloid cell-1 (TREM-1) is a superimmunoglobulin receptor expressed on myeloid cells. Synergy between TREM-1 and Toll-like receptor amplifies the inflammatory response; however, the mechanisms by which TREM-1 accentuates inflammation are not fully understood. In this study, we investigated the role of TREM-1 in a model of LPS-induced lung injury and neutrophilic inflammation. We show that TREM-1 is induced in lungs of mice with LPS-induced acute neutrophilic inflammation. TREM-1 knockout mice showed an improved survival after lethal doses of LPS with an attenuated inflammatory response in the lungs. Deletion of TREM-1 gene resulted in significantly reduced neutrophils and proinflammatory cytokines and chemokines, particularly IL-1ß, TNF-α, and IL-6. Physiologically deletion of TREM-1 conferred an immunometabolic advantage with low oxygen consumption rate (OCR) sparing the respiratory capacity of macrophages challenged with LPS. Furthermore, we show that TREM-1 deletion results in significant attenuation of expression of miR-155 in macrophages and lungs of mice treated with LPS. Experiments with antagomir-155 confirmed that TREM-1-mediated changes were indeed dependent on miR-155 and are mediated by downregulation of suppressor of cytokine signaling-1 (SOCS-1) a key miR-155 target. These data for the first time show that TREM-1 accentuates inflammatory response by inducing the expression of miR-155 in macrophages and suggest a novel mechanism by which TREM-1 signaling contributes to lung injury. Inhibition of TREM-1 using a nanomicellar approach resulted in ablation of neutrophilic inflammation suggesting that TREM-1 inhibition is a potential therapeutic target for neutrophilic lung inflammation and acute respiratory distress syndrome (ARDS).

Triggering receptor expressed on myeloid cells 1 (TREM-1)-mediated Bcl-2 induction prolongs macrophage survival.

Triggering receptor expressed on myeloid cells 1 (TREM-1) is a superimmunoglobulin receptor expressed on myeloid cells that plays an important role in the amplification of inflammation. Recent studies suggest a role for TREM-1 in tumor-associated macrophages with relationship to tumor growth and progression. Whether the effects of TREM-1 on inflammation and tumor growth are mediated by an alteration in cell survival signaling is not known. In these studies, we show that TREM-1 knock-out macrophages exhibit an increase in apoptosis of cells in response to lipopolysaccharide (LPS) suggesting a role for TREM-1 in macrophage survival. Specific ligation of TREM-1 with monoclonal TREM-1 (mTREM-1) or overexpression of TREM-1 with adeno-TREM-1 induced B-cell lymphoma-2 (Bcl-2) with depletion of the key executioner caspase-3 prevents the cleavage of poly(ADP-ribose) polymerase. TREM-1 knock-out cells showed lack of induction of Bcl2 with an increase in caspase-3 activation in response to lipopolysaccharide. In addition overexpression of TREM-1 with adeno-TREM-1 led to an increase in mitofusins (MFN1 and MFN2) and knockdown of TREM-1 decreased the expression of mitofusins suggesting that TREM-1 contributes to the maintenance of mitochondrial integrity favoring cell survival. Investigations into potential mechanisms by which TREM-1 alters cell survival showed that TREM-1-induced Bcl-2 in an Egr2-dependent manner. Furthermore, our data shows that expression of Egr2 in response to specific ligation of TREM-1 is ERK mediated. These data for the first time provide novel mechanistic insights into the role of TREM-1 as an anti-apoptotic protein that prolongs macrophage survival.

Induction of cyclooxygenase-2 signaling by Stomatococcus mucilaginosus highlights the pathogenic potential of an oral commensal.

Stomatococcus mucilaginosus is an oral commensal that has been occasionally reported to cause severe infections in immunocompromised patients. There is no information about the pathogenic role of S. mucilaginosus in airway infections. In a cohort of 182 subjects with bronchiectasis, we found that 9% were colonized with S. mucilaginosus in their lower airways by culture growth from bronchoalveolar lavage. To address the pathogenic potential of S.mucilaginosus, we developed a murine model of S. mucilaginosus lung infection. Intratracheal injection of S. mucilaginosus in C57BL/6 mice resulted in a neutrophilic influx with production of proinflammatory cytokines, chemokines, and lipid mediators, mainly PGE2 with induction of cyclooxygenase-2 (COX-2) in the lungs. Presence of TLR2 was necessary for induction of COX-2 and production of PGE2 by S. mucilaginosus. TLR2-deficient mice showed an enhanced clearance of S. mucilaginosus compared with wild-type mice. Administration of PGE2 to TLR2(-/-) mice resulted in impaired clearance of S. mucilaginosus, suggesting a key role for COX-2-induced PGE2 production in immune response to S. mucilaginosus. Mechanistically, induction of COX-2 in macrophages was dependent on the p38-ERK/MAPK signaling pathway. Furthermore, mice treated with S. mucilaginosus and Pseudomonas aeruginosa showed an increased mortality compared with mice treated with PA103 or S. mucilaginosus alone. Inhibition of COX-2 significantly improved survival in mice infected with PA103 and S. mucilaginosus. These data provide novel insights into the bacteriology and personalized microbiome in patients with bronchiectasis and suggest a pathogenic role for S. mucilaginosus in patients with bronchiectasis.

A survey of mechanical antennas applied for low-frequency transmitting.

The mechanical antenna (MA) is a new type of low-frequency (LF) transmitting antenna that generates an alternating electromagnetic (EM) signal through the mechanical movement of electric charges or magnetic dipoles, which is an interdisciplinary field including not only antennas but also electromagnetics, materials science, and dynamics. This principle of signaling makes it possible to break the constraints on physical dimensions decided by the wavelength of the traditional antenna so as to achieve LF communications with a smaller size and to provide a novel solution for long-range, underwater, and underground communications, navigation over the horizon, and geological exploring. Therefore, MA has become a research hotspot in the field of LF communications in recent 5 years, and this work proposed a survey on this topic of MA applied for LF transmitting. Firstly, we briefly review traditional low-frequency transmitting antennas and summarize the defect; then we introduce research progress of different implementation schemes for MA, comparing the signaling performance, advantages, and disadvantages of each scheme. Furthermore, we discuss the experiment setup, results, and related technology for MA including signal modulation methods. Finally, we explore prospects for future research about MA. This work presents a comprehensive and critical survey of small LF transmitters based on MA to help the readers to understand and identify the background, status, and challenges of research in this field.

Synthesis of CsPbBr3 in Micro Total Reaction System: Fast Operation Space Mapping and Subsecond Growth Process Monitoring.

Lead halide perovskite nanocrystals (LHP NCs) have the characteristics of fast reaction kinetics and crystal instability due to the intrinsically highly ionic bonding between the respective ions, which bring challenges for revealing the growth kinetics and practical applications. Compared with conventional batch synthesis methods, the single-function microreactor can achieve precise and stable control of the NCs synthesis process, but it still has the shortcoming of not being able to obtain information about the growth process. In this study, a micro Total Reaction System (µTRS) with remote control, online detection, and rapid data analysis functions is designed. µTRS can sample the photoluminescence information of CsPbBr3 NCs growth in ligand-assisted reprecipitation method. CsPbBr3 NCs with an emission range of 435-492 nm are successfully detected, which breaks the record of the smallest size of CsPbBr3 NCs synthesized directly from precursors. The real-time feature of µTRS enables the construction of an automated close-loop synthesis system. Besides, the rapid acquisition and timely processing of product information enable the rapid mapping of the operation space for CsPbBr3 NCs preparation, which provides a reliable and learnable data set for designing a fully autonomous microreaction system capable of synthesizing NCs.

Yu-Shiba-Rusinov states assisted by asymmetric Coulomb repulsion in a bipartite molecular device.

Interfacing magnetism with superconducting condensates are promising candidates holding Majorana bound states with which fault-tolerant quantum computation could be implemented. Within this field, understanding the detailed dynamics is important both for fundamental reasons and for the development of innovative quantum technologies. Herein, motivated by a molecular magnet Tb2Pc3interacting with a superconducting Pb(111) substrate, which results in spin-orbital Yu-Shiba-Rusinov (YSR) states, as is affirmed by a theoretical simulation with the aid of the numerical renormalization group technique (see Xiaet al2022Nat. Commun.136388), we study the YSR states and quantum phase transitions (QPTs) in a bipartite molecular device adsorbed on ans-wave superconducting substrate. We highlight the effect of asymmetric Coulomb repulsion by computing the spectrum function and spin correlation function in various parameter regimes. We demonstrate that if one impurity is non-interacting, there are no YSR states in both impurities with any repulsion value in the other impurity. Whereas if the repulsion in one impurity is strong, the YSR states are observed in both impurities, and a QPT arises as the repulsion in the other impurity sweeps, assisted by the competition between the superconducting singlet (Cooper pair) and the Kondo singlet. The evolution of YSR states distinguishes from the single impurity case and can be well interpreted by the energy scales of the isotropic superconducting gap parameter, as well as the two Kondo temperatures. Our findings provide theoretical insights into the phase diagram of two magnetic impurities on a superconducting host and shine light on the effects induced by asymmetric Coulomb repulsion on many-body interactions.

Denitrification Performance in Packed-Bed Reactors Using Novel Carbon-Sulfur-Based Composite Filters for Treatment of Synthetic Wastewater and Anaerobic Ammonia Oxidation Effluent.

To avoid nitrate pollution in water bodies, two low-cost and abundant natural organic carbon sources were added to make up the solid-phase denitrification filters. This study compared four novel solid-phase carbon-sulfur-based composite filters, and their denitrification abilities were investigated in laboratory-scale bioreactors. The filter F4 (mixture of elemental sulfur powder, shell powder, and peanut hull powder with a mass ratio of 6:2.5:1.5) achieved the highest denitrification ability, with an optimal nitrate removal rate (NRR) of 723 ± 14.2 mg NO3 --N⋅L-1⋅d-1 when the hydraulic retention time (HRT) was 1 h. The HRT considerably impacted effluent quality after coupling of anaerobic ammonium oxidation (ANAMMOX) and solid-phase-based mixotrophic denitrification process (SMDP). The concentration of suspended solids (SS) of the ANAMMOX effluent may affect the performance of the coupled system. Autotrophs and heterotrophs were abundant and co-existed in all reactors; over time, the abundance of heterotrophs decreased while that of autotrophs increased. Overall, the SMDP process showed good denitrification performance and reduced the sulfate productivity in effluent compared to the sulfur-based autotrophic denitrification (SAD) process.

Mathematical Modeling for the Industrial 2-Mercaptobenzothiazole Batch Production Process.

As an important chemical intermediate, 2-mercaptobenzothiazole (MBT) is widely used in various processes, especially in the rubber industry. However, there is no first-principles model that describes the synthetic process of MBT. This paper focuses on the formulation of a reliable mathematical model represented by a series of differential and algebraic equations for the industrial batch MBT production process. It is difficult to estimate all of the unknown parameters in the model because of the lack of sufficient industrial/experimental data. Thus, a reduced estimable parameter set is derived by performing estimability analysis on the original estimation problem. A multiple-starting-point strategy is then applied to numerically solve the non-convex parameter estimation problem with the weighted least-squares approach. Subsequently, a cross-validation technique is employed to evaluate the generalizability of the proposed model. Finally, it is confirmed that the proposed model produces encouraging prediction performance with regard to independent test data.

Semiquantitative naked-eye detection of synthetic food colorants using highly-branched pipette tip as an all-in-one device.

The existing strategies for the determination of synthetic food colorants (FCs) in manufactured foods are highly relied on specialized instruments and skilled personnel which are limited by the high technical threshold and instrumentation cost. Herein, highly branched pipette tips (PTs) were fabricated as a robust all-in-one device for high-performance extraction and visual detection of FCs via handy aspiration and dispensing procedures of pipette controller. The density of extraction groups and inner specific surface area of PTs greatly increased after facile physical coating and subsequent layer-by-layer branching reactions, and the maximum increment in binding capacity of PTs was exceeded 300 times at 8-10 iterations of branching layers, enabling the PTs to be colored just by short-time extraction of FCs and to achieve the instrument-independent visual detection of FCs by virtue of their outstanding PT-SPE performance. As a proof-of-concept, the in-situ PT-based solid phase extraction (PT-SPE) with high recoveries (from 91.73 ± 4.76% to 99.90 ± 4.14%) and semiquantitative naked-eye detection of FCs (Allura red and brilliant blue) in real beverages were experimentally demonstrated to be highly feasible by comparison with classical techniques like spectrophotometry, HPLC, and mass spectrometry.

The Effect of PGC-1alpha-SIRT3 Pathway Activation on Pseudomonas aeruginosa Infection.

The innate immune response to P. aeruginosa pulmonary infections relies on a network of pattern recognition receptors, including intracellular inflammasome complexes, which can recognize both pathogen- and host-derived signals and subsequently promote downstream inflammatory signaling. Current evidence suggests that the inflammasome does not contribute to bacterial clearance and, in fact, that dysregulated inflammasome activation is harmful in acute and chronic P. aeruginosa lung infection. Given the role of mitochondrial damage signals in recruiting inflammasome signaling, we investigated whether mitochondrial-targeted therapies could attenuate inflammasome signaling in response to P. aeruginosa and decrease pathogenicity of infection. In particular, we investigated the small molecule, ZLN005, which transcriptionally activates peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), a master regulator of mitochondrial biogenesis, antioxidant defense, and cellular respiration. We demonstrate that P. aeruginosa infection promotes the expression of inflammasome components and attenuates several components of mitochondrial repair pathways in vitro in lung epithelial cells and in vivo in an acute pneumonia model. ZLN005 activates PGC-1α and its downstream effector, Sirtuin 3 (SIRT3), a mitochondrial-localized deacetylase important for cellular metabolic processes and for reactive oxygen species homeostasis. ZLN005 also attenuates inflammasome signaling induced by P. aeruginosa in bronchial epithelial cells and this action is dependent on ZLN005 activation of SIRT3. ZLN005 treatment reduces epithelial-barrier dysfunction caused by P. aeruginosa and decreases pathogenicity in an in vivo pneumonia model. Therapies that activate the PGC-1α-SIRT3 axis may provide a complementary approach in the treatment of P. aeruginosa infection.

Colorimetry Combined with Inner Filter Effect-Based Fluorometry: A Versatile and Robust Strategy for Multimode Visualization of Food Dyes.

Herein, a strategy combining colorimetry and inner filter effect (IFE)-based fluorometry was developed for multimode visualization of food dyes (FDs) using CdTe quantum-dots-doped fluorescent indicator papers as a sample-to-answer device. Colorimetry was straightforwardly achieved by FDs extraction through electrostatic interaction and hydrophobic effect while fluorometry was implemented by IFE-induced fluorescence quenching. RGB/gray-scale values of colorimetry and fluorometry were furtherly picked by a smartphone application and applied to reconstruct color information-based digital image analysis for both direct alignments and linear regression analysis. The apparent color and fluorescence of FDs-bound indicator papers, together with their digitized color information, showed a good mapping to FDs concentrations in the range of 0-0.5 mg/mL for Sunset Yellow, 0-0.2 mg/mL for Allura Red, and 0-0.08 mg/mL for Brilliant Blue. As a proof of concept, the dosages of these FDs in real beverages and simulated dye effluents were deduced and cross-validated by different visualization modes, and finally double-checked by instrumental techniques such as spectrometric methods, high-performance liquid chromatography (HPLC), and mass spectroscopy (MS). The above findings concluded that (i) IFE mechanism is generally applicable to build fluorometric systems and (ii) cross validation of different visualization modes can markedly improve detection accuracy, which may provide references for design and fabrication of novel "lab-on-paper" devices for visualization applications with high reliability.

Immunoproteomic analysis of the antibody response obtained in mouse following vaccination with a T-cell vaccine.

T-cell vaccination (TCV), the application of irradiated activated T cells, has been shown to prevent effectively and treat experimental autoimmune diseases. It has been reported that anti-lymphocytic antibodies induced by TCV were capable of strongly inhibiting T-cell proliferation and of ameliorating experimental autoimmune disease. The present study was undertaken to characterize the antigen specificity of these Abs. We used activated mouse ovalbumin (OVA)-specific T cells (OVA-T) as vaccine immunized mice. By combination of 2-DE, 2-D Western blot and Q-TOF mass spectrometry we have identified 11 antigens in activated T cells that were recognized by the anti-T-cell Abs. The resulting antigenic molecules included calreticulin (CRT), ERp57, Vimentin, HSP70-4, tubulin ß5 chain, coronin-1A, pyruvate kinase, ATP synthase ß chain and transketolase most of which belong to so-called damage-associated molecular pattern molecules (DAMPs). CRT, ERp57 and vementin were further examined by Western blot and cellular ELISA to identify molecular targets which may be involved in the TCV immunotherapy. On the basis of our results, γ-radiation induced the activated T cells "immunogenic apoptosis" and exposed/secreted DAMPs (CRT, ERp57 and Vementin) played an important role in TCV therapy.

MicroRNA-155 Modulates Macrophages' Response to Non-Tuberculous Mycobacteria through COX-2/PGE2 Signaling.

Non-tuberculous mycobacteria (NTM) have been recognized as a causative agent of various human diseases, including severe infections in immunocompromised patients, such as people living with HIV. The most common species identified is the Mycobacterium avium-intracellulare complex (MAI/MAC), accounting for a majority of infections. Despite abundant information detailing the clinical significance of NTM, little is known about host-pathogen interactions in NTM infection. MicroRNAs (miRs) serve as important post-transcriptional regulators of gene expression. Using a microarray profile, we found that the expression of miR-155 and cyclo-oxygenase 2 (COX-2) is significantly increased in bone-marrow-derived macrophages from mice and human monocyte-derived macrophages from healthy volunteers that are infected with NTM. Antagomir against miR-155 effectively suppressed expression of COX-2 and reduced Prostaglandin E2(PGE2) secretion, suggesting that COX-2/PGE2 expression is dependent on miR-155. Mechanistically, we found that inhibition of NF-κB activity significantly reduced miR-155/COX-2 expression in infected macrophages. Most importantly, blockade of COX-2, E-prostanoid receptors (EP2 and EP4) enhanced killing of MAI in macrophages. These findings provide novel mechanistic insights into the role of miR-155/COX-2/PGE2 signalling and suggest that induction of these pathways enhances survival of mycobacteria in macrophages. Defining host-pathogen interactions can lead to novel immunomodulatory therapies for NTM infections which are difficult to treat.