Comparative Analysis of In situ Fibronectin Using ToF-SIMS, SPI-MS, and dropDESI-MS in a Microfluidic Reactor

Fibronectin is an important biomolecule due to its role in cell differentiation, growth, kinesis, and adhesion. Such biological responses are mediated through membrane recognition and signaling; where fibronectin is found. Studying the outer molecular surface of fibronectin allows deeper insight into the microbiological reactions that occur during these processes. In situ mass spectrometry analysis in aqueous solution accurately represents fibronectin’s chemical components, made possible by a vacuum compatible microfluidic reactor, SALVI (System for Analysis at the Liquid Vacuum Interface). SALVI was paired with the analytical tools: time-of-flight secondary ion mass spectrometer (ToF-SIMS), single photon ionization mass spectrometer (SPI-MS) and drop desorption electrospray ionization mass spectrometer (dropDESI-MS). ToF-SIMS employed a bismuth liquid metal ion beam. Positive and negative ion spectral plots were constructed and analyzed. The advanced light source (ALS) SPI-MS), using a synchrotron vacuum ultraviolet (VUV) light, elicited data depending on varying photoionization efficiencies (PIE). PIE plots were examined for the initial detection of photons of a mass to charge ratio (m/z), resulting in the determination of the ionization efficiency (IE) of a corresponding compound. Both ToF-SIMS and SPI-MS are surface tools, with ion beam impact no further than the second monolayer. DropDESI-MS, analyzed under ambient conditions, utilized a capillary connecting the electrode spray to the mass spectrometer. Charged microdroplets were used to introduce samples to the mass analyzer. Central masses (m/z) from all three apparatuses were identified to their most possible compounds or constituents, demonstrating complementary results. Mass identifications were based on literature survey and results from peer reviewed articles. Our results suggest the need for further research of organic compounds, like fibronectin, to understand their surface compositions in aqueous solution

Salvia miltiorrhiza injection ameliorates myocardial ischemia-reperfusion injury via downregulation of PECAM-1

Purpose: To investigate the effect of Salvia miltiorrhiza injection on myocardial ischemia-reperfusion injury and PECAM-1 related pathways. Method: Male Wistar rats were used for establishment of myocardial ischemia-reperfusion model. The rats were randomly assigned to four groups: experimental group, low dose group (Salvia miltiorrhiza injection, 10 mL/kg/day), moderate dose group (Salvia miltiorrhiza injection, 20 mL/kg/day) and high dose group (Salvia miltiorrhiza injection, 40 mL/kg/day). Myocardial ischemia-reperfusion model was established in the four groups. Evans-TTC staining was used to assess relative area of ischemiareperfusion injury. Blood samples were collected for assay of PECAM-1 expression using enzymelinked immunosorbent assay (ELISA). Fresh blood platelets were collected in all groups, and divided into two groups - control group (normal culture) and experimental group (Salvia miltiorrhiza injection). The expression of PECAM-1 in blood platelets was assayed using Western blot. Result: Compared with the experimental group, Salvia miltiorrhiza injection ameliorated myocardial ischemia-reperfusion injury, and decreased the infarction area seen in Evans/TTC staining. PECAM-1 expression in blood was decreased by Salvia miltiorrhiza injection. Blood platelets dysfunction was induced after myocardial ischemia-reperfusion, and the level of PECAM-1 increased. However, Salvia miltiorrhiza injection treatment downregulated the expression of PECAM-1 after myocardial ischemiareperfusion. Conclusion: Salvia miltiorrhiza injection maintains normal function of blood platelets and ameliorates myocardial ischemia-reperfusion injury by decreasing expression of PECAM-1

Using In Situ Liquid Single Photon Ionization Mass Spectrometry (SPI-MS) to Probe Lithium Polysulfide Electrolyte in Motion

The solid-liquid (s-l) interface is the most common interface encountered in electrochemical systems. The s-l interface has wide applications in energy storage, catalysis, and material sciences. In situ studies of chemical reactions taking place on the s-l interfaces can further our understanding of electron transfer and link to real-world device functions under challenging conditions. Direct probing of the solid electrode and liquid electrolyte interface has been realized using a vacuum compatible electrochemical microfluidic reactor, system for analysis at the liquid vacuum interface (SALVI) with time-of-flight secondary ion mass spectrometry (ToF-SIMS). Most recently, the electrochemical version of SALVI was integrated to the synchrotron based single photon ionization mass spectrometry (SPI-MS). SPI-MS has proven to be a versatile technique for analysis of organic species in the solid or gas phase due to its nature of soft ionization. As a practical example, three different lithium polysulfide electrolytes, Li2S4, Li2S6, and Li2S8, were studied under dynamic conditions with various applied voltages. It was found that despite some PDMS interference peaks such as 369 m/z, unique peaks of interest signifying the electron transfer of the LixSy electrolytes can be identified according to the SPI-MS mass spectra. The observation of in situ compositional changes as a result of electrochemical reaction that take place at the s-l interface in a three electrode system allowed us to piece all the fragments together and identify the compound present in the sample at different stages of photoionization energy (PIE) values. Moreover, we demonstrate that liquid SPI-MS technique has been enabled to study dynamic electron transfer of LixSy electrolytes using real-time molecular imaging

Icariin Protects Bone Marrow Mesenchymal Stem Cells Against Iron Overload Induced Dysfunction Through Mitochondrial Fusion and Fission, PI3K/AKT/mTOR and MAPK Pathways

Iron overload has been reported to contribute to bone marrow mesenchymal stem cells (BMSCs) damage, but the precise mechanism still remains elusive. Icariin, a major bioactive monomer belonging to flavonoid glucosides isolated from Herba Epimedii, has been shown to protect cells from oxidative stress induced apoptosis. The aim of this study was to investigate whether icariin protected against iron overload induced dysfunction of BMSCs and its underlying mechanism. In this study, we found that iron overload induced by 100 μM ferric ammonium citrate (FAC) caused apoptosis of BMSCs, promoted cleaved caspase-3 and BAX protein expressions while inhibited Bcl-2 protein expression, which effects were significantly attenuated by icariin treatment. In addition, iron overload induced significant depolarization of mitochondrial membrane potential (MMP), reactive oxygen species (ROS) generation and inhibition of mitochondrial fusion/fission, which effects were also attenuated by icariin treatment. Meanwhile, we found that iron overload induced by 100 μM FAC significantly inhibited mitochondrial fission protein FIS1 and fusion protein MFN2 expressions, inhibited DRP1 and Cytochrome C protein translocation from the cytoplasm to mitochondria. Icariin at concentration of 1 μM was able to promote mitochondrial fission protein FIS1 and fusion protein MFN2 expressions, and increase DRP1 and cytochrome C protein translocation from the cytoplasm to mitochondria. Further, osteogenic differentiation and proliferation of BMSCs was significantly inhibited by iron overload, but icariin treatment rescued both osteogenic differentiation and proliferation of BMSCs. Further studies showed that icariin attenuated iron overload induced inactivation of the PI3K/AKT/mTOR pathway and activation of the ERK1/2 and JNK pathways. In summary, our study indicated that icariin was able to protect against iron overload induced dysfunction of BMSCs. These effects were potentially related to the modulation of mitochondrial fusion and fission, activation of the PI3K/AKT/mTOR pathway and inhibition of ERK1/2 and JNK pathways

Process Optimization of Electrochemical Treatment of COD and Total Nitrogen Containing Wastewater

In this work, an electrochemical method for chemical oxygen demand (COD) and total nitrogen (TN, including ammonia, nitrate, and nitrite) removal from wastewater using a divided electrolysis cell was developed, and its process optimization was investigated. This process could effectively relieve the common issue of NO3−/NO2− over-reduction or NH4+ over-oxidation by combining cathodic NO3−/NO2− reduction with anodic COD/NH4+ oxidation. The activity and selectivity performances toward pollutant removal of the electrode materials were investigated by electrochemical measurements and constant potential electrolysis, suggesting that Ti electrode exhibited the best NO3−/NO2− reduction and N2 production efficiencies. In-situ Fourier transform infrared spectroscopy was used to study the in-situ electrochemical information of pollutants conversion on electrode surfaces and propose their reaction pathways. The effects of main operating parameters (i.e., initial pH value, Cl− concentration, and current density) on the removal efficiencies of COD and TN were studied. Under optimal conditions, COD and TN removal efficiencies from simulated wastewater reached 92.7% and 82.0%, respectively. Additionally, reaction kinetics were investigated to describe the COD and TN removal. Results indicated that COD removal followed pseudo-first-order model; meanwhile, TN removal followed zero-order kinetics with a presence of NH4+ and then followed pseudo-first-order kinetics when NH4+ was completely removed. For actual pharmaceutical wastewater treatment, 79.1% COD and 87.0% TN were removed after 120 min electrolysis; and no NH4+ or NO2− was detected

Process Optimization of Electrochemical Oxidation of Ammonia to Nitrogen for Actual Dyeing Wastewater Treatment

To mitigate the potential environmental risks caused by nitrogen compounds from industrial wastewater, residual ammonia after conventional wastewater treatment should be further eliminated. In this work, an electrochemical oxidation process for converting ammonia to nitrogen in actual dyeing wastewater was investigated. The effects of the main operating parameters, including initial pH value, applied current density, NaCl concentration, and flow, were investigated on ammonia removal and products distribution. Experimental results indicated that, under optimal conditions of an initial pH value of 8.3, applied current density of 20 mA cm−2, NaCl concentration of 1.0 g L−1, and flow of 300 mL min−1, the ammonia could be completely removed with N2 selectivity of 88.3% in 60 min electrolysis. A kinetics investigation using a pseudo-first-order model provided a precise description of ammonia removal during the electro-oxidation process. Experimental functions for describing the relationships between kinetic constants of ammonia removal and main operating parameters were also discussed. Additionally, the mechanisms and economic evaluation of ammonia oxidation were conducted. All these results clearly proved that this electro-oxidation process could efficiently remove ammonia and achieve high N2 selectivity

Deferoxamine Alleviates Osteoarthritis by Inhibiting Chondrocyte Ferroptosis and Activating the Nrf2 Pathway

Objective: Osteoarthritis (OA) is a common disease with a complex pathology including mechanical load, inflammation, and metabolic factors. Chondrocyte ferroptosis contributes to OA progression. Because iron deposition is a major pathological event in ferroptosis, deferoxamine (DFO), an effective iron chelator, has been used to inhibit ferroptosis in various degenerative disease models. Nevertheless, its OA treatment efficacy remains unknown. We aimed to determine whether DFO alleviates chondrocyte ferroptosis and its effect on OA and to explore its possible mechanism.Methods: Interleukin-1β (IL-1β) was used to simulate inflammation, and chondrocyte ferroptosis was induced by erastin, a classic ferroptosis inducer. A surgical destabilized medial meniscus mouse model was also applied to simulate OA in vivo, and erastin was injected into the articular cavity to induce mouse knee chondrocyte ferroptosis. We determined the effects of DFO on ferroptosis and injury-related events: chondrocyte inflammation, extracellular matrix degradation, oxidative stress, and articular cartilage degradation.Results: IL-1β increased the levels of ROS, lipid ROS, and the lipid peroxidation end product malondialdehyde (MDA) and altered ferroptosis-related protein expression in chondrocytes. Moreover, ferrostatin-1 (Fer-1), a classic ferroptosis inhibitor, rescued the IL-1β–induced decrease in collagen type II (collagen II) expression and increase in matrix metalloproteinase 13 (MMP13) expression. Erastin promoted MMP13 expression in chondrocytes but inhibited collagen II expression. DFO alleviated IL-1β– and erastin-induced cytotoxicity in chondrocytes, abrogated ROS and lipid ROS accumulation and the increase in MDA, improved OA-like changes in chondrocytes, and promoted nuclear factor E2–related factor 2 (Nrf2) antioxidant system activation. Finally, intra-articular injection of DFO enhanced collagen II expression in OA model mice, inhibited erastin-induced articular chondrocyte death, and delayed articular cartilage degradation and OA progression.Conclusion: Our research confirms that ferroptosis occurs in chondrocytes under inflammatory conditions, and inhibition of chondrocyte ferroptosis can alleviate chondrocyte destruction. Erastin-induced chondrocyte ferroptosis can stimulate increased MMP13 expression and decreased collagen II expression in chondrocytes. DFO can suppress chondrocyte ferroptosis and promote activation of the Nrf2 antioxidant system, which is essential for protecting chondrocytes. In addition, ferroptosis inhibition by DFO injection into the articular cavity may be a new OA treatment

M Protein of Group a Streptococcus Plays an Essential Role in Inducing High Expression of A20 in Macrophages Resulting in the Downregulation of Inflammatory Response in Lung Tissue

Group A streptococcus (GAS), a common pathogen, is able to escape host immune attack and thus survive for longer periods of time. One of the mechanisms used by GAS is the upregulated expression of immunosuppressive molecules, which leads to a reduction in the production of inflammatory cytokines in immune cells. In the present study, we found that macrophages produced lower levels of proinflammatory cytokines (IL-1β, TNF-α, IL-6) when challenged with GAS than they did when challenged with Escherichia coli (E. coli). Simultaneously, in a mouse model of lung infection, GAS appeared to induce a weaker inflammatory response compared to E. coli. Our data also indicated that the expression of the A20 transcriptional regulator was higher in GAS-infected macrophages than that in macrophages infected with E. coli, and that high expression of A20 correlated with a reduction in the production of TRAF6. SiRNA targeting of A20 led to the increased production of TRAF6, IL-1β, TNF-α, and IL-6, suggesting that A20 inhibits synthesis of these key proinflammatory cytokines. We also investigated the pathway underlying A20 production and found that the synthesis of A20 depends on My88, and to a lower extent on TNFR1. Finally, we showed a significant reduction in the expression of A20 in macrophages stimulated by M protein-mutant GAS, however, a speB-GAS mutant, which is unable to degrade M protein, induced a greater level of A20 production than wild type GAS. Collectively, our data suggested that M protein of GAS was responsible for inducing A20 expression in macrophages, which in turn down-regulates the inflammatory cytokine response in order to facilitate GAS in evading immune surveillance and thus prolong survival in the host

DataSheet2_Genes related to N6-methyladenosine in the diagnosis and prognosis of idiopathic pulmonary fibrosis.XLS

Introduction: Idiopathic pulmonary fibrosis (IPF) is a chronic progressive pulmonary fibrotic disease with unknown etiology and poor outcomes. It severely affects the quality of life. In this study, we comprehensively analyzed the expression of N6-methyladenosine (m6A) RNA methylation regulators using gene expression data from various tissue sources in IPF patients and healthy volunteers.Methods: The gene expression matrix and clinical characteristics of IPF patients were retrieved from the Gene Expression Omnibus database. A random forest model was used to construct diagnosis signature m6A regulators. Regression analysis and correlation analysis were used to identify prognosis m6A regulators. Consensus cluster analysis was used to construct different m6A prognosis risk groups, then functional enrichment, immune infiltration and drug sensitivity analysis were performed.Result: Five candidate m6A genes from lung tissue were used to predict the incidence, and the incidence was validated using datasets from bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells. Subsequently, the BALF dataset containing outcomes data was used for the prognosis analysis of m6A regulators. METTL14, G3BP2, and ZC3H13 were independent protective factors. Using correlation analysis with lung function in the lung tissue-derived dataset, METTL14 was a protective factor in IPF. Based on METTL14 and G3BP2, a consensus cluster analysis was applied to distinguish the prognostic m6A regulation patterns. The low-risk group’s prognosis was significantly better than the high-risk group. Biological processes regulated by various risk groups included fibrogenesis and cell adhesion. Analysis of immune cell infiltration showed upregulation of neutrophils in the m6A high-risk group. Subsequently, five m6A high-risk group sensitive drugs and one m6A low-risk group sensitive drug were identified.Discussion: These findings suggest that m6A regulators are involved in the diagnosis and prognosis of IPF, and m6A patterns are a method to identify IPF outcomes.</p